2010 COLD SPRING HARBOR LABORATORY CONFERENCE ON MOLECULAR CHAPERONES AND STRESS
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 5R13AG037223-02 from National Institute On Aging
Abstract: This proposal is a request for partial financial support for a meeting on Molecular Chaperones and Stress Responses to be held from May 4 - 8, 2010 at Cold Spring Harbor Laboratory. This meeting is the premier international format for presentation of new results in this area, and is attended by representatives from virtually every major laboratory in the field. The explosion of new information on how the folded state of proteins is acquired and maintained in vivo and the involvement of this process in an increasing number of disease states and in normal aging guarantees the excitement of this meeting. Among the highlights of the meeting will be sessions devoted to (1) Protein misfolding and disease, emphasizing the role of alternative folding pathways in protein biogenesis and lifecycle and mounting evidence for chaperone involvement in Alzheimer´s Disease, polyglutamine diseases and the aging process (2) new structural and mechanistic information on some intensively studied chaperones, (3) the role of chaperones in diverse cellular transactions and the signal transduction pathways that integrate them, (4) the crosstalk between protein folding and proteolysis mediated by chaperones, (5) how the various protein folding and quality control systems in a cell function together to provide a robust protein folding environment and how the breakdown of such protein homeostasis (proteostasis) contributes to disease and normal aging, (6) the diverse protein quality control strategies used by a cell to ensure the integrity of the secretory pathway during times of protein folding stress. The field of heat shock proteins and molecular chaperones has grown rapidly and draws interest not only from traditional scientific disciplines in the basic sciences but also from numerous areas of biomedical research including neurodegenerative disease, infectious diseases, cancer, heart disease and aging. The meeting will include seven lecture and three poster sessions. The proposed sessions include I- Chaperone Biochemistry & Protein Folding, II- Chaperone Function in Disease and Development, III- Diseases of Protein Misfolding IV- Quality Control & Protein Trafficking V- Evolution and Regulation of Protein Folding Machines, VI- Chaperones and Proteolysis, and VII- Manipulating Chaperone Networks and Protein Folding Pathways. Each session will consist of eight to nine oral presentations and will be chaired by an invited speaker. A maximum of two additional speakers will be pre-invited per session and the remainder will be selected from submitted s. This balance of talks allows the meeting to feature presentations by leading scientists, to be responsive to exciting new developments, to encourage diverse participation and to recognize new investigators. The subsequent meetings (2012 and 2014) will follow a similar format and will include topics highly relevant to the current research at the time of the meeting. Proteins are composed of polymers of amino acids (or polypeptides) and execute essential tasks in all living organisms. Protein function is critically dependent on the folding of their constituent polypeptides into active three-dimensional objects and experimental evidence accumulated in the recent past has drawn attention to contribution that failure of this process (protein misfolding) makes to some of the most important diseases affecting humanity. The latter include common neurodegenerative disorders, such as Parkinson´s disease and Alzheimer´s disease, common metabolic disorders such as Diabetes Mellitus and a host of other conditions liked to the aging process. The Cold Spring Harbor Meeting on Molecular Chaperones and Stress Responses brings together experts in diverse aspects of protein folding and the cellular response to protein misfolding. As such it serves as the premier clearinghouse for ideas on how to eventually translate the basic discoveries in the area to future treatments of recalcitrant diseases
Keywords: ing; Affect; Aging; Aging-Related Process; Alzheimer`s Disease; Amino Acids; Area; Attention; Basic Science; Biochemistry; Biogenesis; biological adaptation to stress; Biomedical Research; Cell physiology; Cells; Communicable Diseases; Development; Diabetes Mellitus; Discipline; Disease; Ensure; Environment; Equilibrium; Evolution; Explosion; Failure (biologic function); Financial Support; Future; Heart Diseases; Heat shock proteins; Homeostasis; Humanities; in vivo; interest; International; Laboratories; lectures; Life; Malignant Neoplasms; Mediating; meetings; Metabolic Diseases; Molecular Chaperones; Neurodegenerative Disorders; normal aging; Oral; Organism; Parkinson Disease; Pathway interactions; polyglutamine; Polymers; polypeptide; posters; Process; protein folding; protein function; protein misfolding; protein transport; Proteins; Proteolysis; public health relevance; Quality Control; Regulation; Research; Research Personnel; response; Role; Scientist; Signal Transduction Pathway; Stress; symposium; System; Time; Translating
Project start date: 2010-05-01
Project end date: 2013-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-08-149
5R13AG037223-02 (2011): $1
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to J David
REGULATION OF CELLULAR ZINC HOMEOSTASIS
J David, Professor
University Of Wisconsin Madisoncity: Madison country: United States (us)
Grant 5R01GM056285-15 from National Institute Of General Medical Sciences
Abstract: Zinc is an essential nutrient because of the important roles this metal plays as a catalytic and structural cofactor. Several zinc-dependent proteins reside within organelles such as the endoplasmic reticulum (ER), Golgi, and mitochondria. Therefore, transporter proteins are needed to distribute zinc into intracellular compartments. Because excess zinc can be toxic, cells also require homeostatic mechanisms to control the intracellular levels of free or labile zinc in the cytosol and within organelles. Cellular zinc homeostasis is achieved by several mechanisms including the control of zinc uptake, efflux, vesicular storage, and binding by metallothionein. In this proposal, another important facet of zinc homeostasis is considered, i.e. the control of zinc levels within intracellular organelles. These processes will be studied using the yeast Saccharomyces cerevisiae as a model eukaryotic cell. Preliminary results have raised five central hypotheses that provide the foundation for the specific aims of this proposal 1) The yeast Msc2 and Zrg17 proteins form a heteromeric complex that transports zinc into the ER. In Aim 1, the composition of this complex will be assessed and its zinc transport activity will be characterized. 2) It is proposed that zinc finger domains will make useful probes of zinc status in the ER and other compartments of living cells. In Aim 2, new in vivo zinc sensors will be developed based on fluorescence resonance energy transfer (FRET). These and other assays to be developed will be used to define the role of Msc2/Zrg17 and other transporters in maintaining ER zinc homeostasis. 3) Zinc transport into the ER via the Msc2/Zrg17 complex is regulated in response to zinc status by both transcriptional and post-translational control mechanisms. In Aim 3, the molecular mechanisms of these regulatory systems will be dissected and their roles in maintaining ER zinc status determined. 4) It is proposed that zinc transport into the ER is mediated by transporters in addition to the Msc2/Zrg17 complex. In Aim 4, other zinc transporters for this compartment will be identified. 5) Several studies of mammalian cells showed that zinc deficiency leads to increased oxidative stress, the source of which has long been a mystery. In Aim 5, the hypothesis that the oxidative stress of zinc deficiency arises from the loss of ER zinc homeostasis will be tested. These aims represent a cohesive and comprehensive analysis of zinc homeostasis in the secretory pathway of eukaryotic cells.7. The processes of intracellular zinc transport and homeostasis within organelles are essential for basic cellular function, physiology, and human health. Despite this importance, however, we know little about the transporters involved and how they are regulated in response to zinc status and other factors. As a result of our proposed studies, we will obtain a fundamental understanding of zinc metabolism in the secretory pathway of all eukaryotes and generate new probes of intracellular zinc that will be useful in our studies as well as in the analysis of zinc metabolism in mammals and other organisms
Keywords: base; Binding (Molecular Function); Biological Assay; Carrier Proteins; cell growth regulation; Cell physiology; Cells; cofactor; Complex; Cytosol; Endoplasmic Reticulum; Environment; Eukaryota; Eukaryotic Cell; extracellular; Fingers; Fluorescence Resonance Energy Transfer; Foundations; Golgi Apparatus; Growth; Health; Homeostasis; Human; in vivo; Life; Mammalian Cell; Mammals; Mediating; Metabolism; Metallothionein; Metals; Mitochondria; Modeling; Molecular; Nutrient; Organelles; Organism; Oxidative Stress; Pathway interactions; Physiology; Play; Process; Proteins; response; Role; Saccharomyces cerevisiae; sensor; Source; System; Testing; uptake; Yeasts; Zinc; Zinc deficiency; Zinc Fingers; zinc-binding protein
Project start date: 1997-09-30
Project end date: 2012-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-07-070
5R01GM056285-15 (2011): $317407
THE DEVELOPMENTAL ORIGINS OF DISEASE AND DETERIORATION IN OLD AGE
J David, Adjunct Professor
Oregon Health And Science Universitycity: Portland country: United States (us)
Grant 5R01AG032339-02 from National Institute On Aging
Abstract: Within any level of socio-economic status there is a wide variation in lifespan. Our hypothesis is that this variation is programmed during development. We propose that poor intrauterine nutrition and poor childhood living conditions initiate pathological processes and permanently change the body´s structure and function in ways that make it vulnerable to stressors in postnatal life. Because intrauterine nutrition is determined by the mother´s lifetime nutrition our findings will quantify how investment in the nutrition and growth of girls and young women will increase the lifespan of the next generation. The Helsinki Birth Cohort Study of 20,000 people is the only known data set in the world that includes maternal body size, birth size, infant and childhood growth and living conditions of people now at the end of their lives. The cohort includes detailed socioeconomic data recorded at each 5 yearly national census. We also propose that the ability to sustain physical and mental health in old age is programmed during development. We will explore the effects of intrauterine nutrition and childhood living conditions on resilience in old age; and we will examine whether there are periods of growth that are critical for this. This will inform preventive strategies that promote well being in early life and protect vulnerable elderly people. In subsequent studies, which are not the subject of this application, we will use the genome wide scan that has already been carried out to distinguish environmental and genetic effects on resilience in old age. Specific Aim 1 To determine how maternal nutrition and childhood living conditions predict all cause mortality, taking into account socio-economic conditions in later life. In two register-based studies of 7086 and 13,345 men and women, we will relate these early life influences to mortality from all causes and from cardiovascular and non-cardiovascular causes separately. Specific Aim 2 To determine how maternal nutrition and childhood living conditions predict changes in physical and mental health in later life, taking into account adult life-style, socio-economic conditions and illness. In a longitudinal study of a sub-sample of 1600 people we will relate these early life influences to changes in indices of physiological, mental and biochemical health. Specific Aim 3 To determine whether there are critical periods of growth between birth and age eleven years that predict all-cause mortality and changes in physical and mental health in later life. In a register- based study of 13,345 men and women, and in a longitudinal study in a sub-sample of 1600 people, we will examine how change in height and body mass index between different ages is related to mortality from cardiovascular and non-cardiovascular causes and to indices of physiological, mental and biochemical health. We propose that the maternal and early postnatal environments establish vulnerability to chronic disease in old age, and determine elderly people´s ability to sustain their health. An understanding of this may make it possible to identify elderly people who are at risk of physical or mental deterioration, and to prolong healthy life
Keywords: Accounting; Admission activity; Adult; Affect; Age; Age at Menarche; American; Animals; base; Behavior; Biochemical; Biological Process; Birth; Birth Records; Blood specimen; Body mass index; Body Size; Breast Feeding; Cardiovascular Diseases; Cardiovascular system; Censuses; Cessation of life; Child; Childhood; Chronic Disease; cohort; Cohort Studies; Competence; Conceptions; Coronary heart disease; Country; critical period; Data; Data Set; Deterioration; Development; developmental plasticity; Diabetes Mellitus; Diet; Disease; DNA; Economic Conditions; Elderly; Environment; Epigenetic Process; Fathers; Fatty acid glycerol esters; Fetal Growth; Fetal Malnutrition; Fetus; fetus nutrition; Finland; Genes; Genetic; genome wide association study; Genotype; girls; Growth; Head circumference; Health; Health Benefit; Healthcare; Height; Home environment; Hormonal; Hospitals; Human; Human Development; Income; indexing; Individual; Infant; Infection; insight; Investments; Length; Life; Life Expectancy; Life Style; Link; Longevity; Longitudinal Studies; Low Birth Weight Infant; Malignant Neoplasms; Malnutrition; Measurement; Measures; men; Menarche; Mental Health; Metabolic; Metabolism; Molecular; Mortality Vital Statistics; mother nutrition; Mothers; Newborn Infant; next generation; Nutrient; nutrition; Occupations; Pathologic Processes; Persons; Pharmaceutical Preparations; Phenotype; physical conditioning; Physiological; Placenta; Plastics; postnatal; Pregnancy; Prevention strategy; programs; Proteins; Psyche structure; public health relevance; Records; Recurrence; Relative (related person); Research; resilience; response; Risk; Sampling; Schools; Signal Transduction; Socioeconomic Status; socioeconomics; stressor; Structure; Uterus; Variant; Weight; Woman
Relevance: Public health relevance We propose that the maternal and early postnatal environments establish vulnerability to chronic disease in old age, and determine elderly people´s ability to sustain their health. An understanding of this may make it possible to identify elderly people who are at risk of physical or mental deterioration, and to prolong healthy life
Project start date: 2010-09-01
Project end date: 2013-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-07-070
5R01AG032339-02 (2011): $167249
TRAVELING WAVES IN VISUAL CORTEX DURING BINOCULAR RIVALRY
J David, Professor
New York Universitycity: New York country: United States (us)
Grant 5R01EY016752-05 from National Eye Institute
Abstract: When the two eyes view dissimilar patterns, one experiences a perceptual phenomenon called binocular rivalry. Instead of seeing both patterns superimposed, they are perceived in alternation. What makes this phenomenon remarkable is the dissociation between a constant physical stimulation and fluctuating perceptual experience. Because of this dissociation, binocular rivalry presents an opportunity for studying visual awareness, one of the deepest mysteries facing biomedical science. In spite of widespread interest, and an impressive volume of high-quality work on this topic, many of the central questions concerning the neural processing underlying binocular rivalry remain open. Particularly controversial is the role of primary visual cortex (V1) in rivalry. To address this controversy, we propose to capitalize on an interesting aspect of the perceptual phenomenon; during an alternation, one sees a traveling wave in which the dominance of one pattern emerges locally and expands progressively as it renders the other pattern invisible. Our experiments are designed to measure and characterize the neural basis of these perceptual waves. The proposed experiments will apply a combination of empirical methods (psychophysics and fMRI in humans; optical imaging, electrophysiology, pharmacology, and electrical stimulation in the awake monkey) to explain this perceptual phenomenon in terms of the underlying neural mechanisms and to test the following hypotheses (1) that competition between the two rival stimuli is implemented by neural circuits in primary visual cortex (V1), i.e., that neural circuits in V1 play a causal role in triggering transitions during rivalry; (2) that for the consequences of this neural competition to be perceived, activity must advance to higher visual areas; and (3) that attention, mediated by feedback from higher visual areas, plays a crucial role in promoting neural activity from V1 to higher visual areas. We will then be in a position to develop and refine a computational theory of the neural processing in V1 that supports these traveling waves, and a theory that elaborates the role of V1 in visual awareness. The neural competition underlying perceptual alternations during rivalry is believed to be closely related to the strabismic suppression. Hence, the proposed experiments will provide useful information and will establish novel experimental protocols that can, in future work; be applied to further our understanding of strabismus and amblyopia
Keywords: Address; Amblyopia; Area; Area striata; area striata; Area striata structure; Attention; awake; Awareness; Awarenesses; base; Binocular rivalry; Coloring Agents; computational modeling; computational models; computational simulation; computer based models; Computer Simulation; computerized modeling; Computerized Models; computerized simulation; Conscious; Consciousness; Cortex, Striate; design; designing; Dissociation; Dominance, Ocular; Dyes; Electric Stimulation; electrical microstimulation; Electrical Stimulation; Electrodes, Miniaturized; Electrophysiology; Electrophysiology (science); experience; experiment; experimental research; experimental study; extracellular; extrastriate visual cortex; Eye; Eye Dominance; Eyeball; Feedback; fMRI; Functional Magnetic Resonance Imaging; Future; Human; Human, General; Image; imaging; in silico; interest; Investigators; Link; Magnetic Resonance Imaging, Functional; Man (Taxonomy); Man, Modern; Maps; Mathematical Model Simulation; Mathematical Models and Simulations; Measurement; Measures; Mediating; Methods; Methods and Techniques; Methods, Other; Microelectrodes; microstimulation; Models, Computer; Monitor; Monkeys; MRI, Functional; Nervous; neural; neural circuit; neural circuitry; neural mechanism; neuromechanism; Neurophysiology / Electrophysiology; novel; Ocular Dominance; optic imaging; optical imaging; Pattern; Perception; Pharmacology; Phorias; Physical Stimulation; Physiologic; Physiological; Physiology; Play; Population; Position; Positioning Attribute; Primary visual cortex; Process; Protocol; Protocols documentation; psycho-physiological; Psychology, Physiologic; Psychology, Physiological; Psychophysic; Psychophysics; Psychophysiological; Psychophysiology; relating to nervous system; Research Personnel; research study; Researchers; Role; Science; Series; Simulation, Computer based; social role; spatiotemporal; Speed; Speed (motion); Squint; Stimulus; Strabismus; Striate area; Techniques; Testing; theories; Travel; V (voltage); V1 neuron; virtual simulation; Visual; Visual Cortex; visual cortical; voltage; Work
Project start date: 2005-09-02
Project end date: 2011-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2011
5R01EY016752-05 (2009): $351475
FUNCTIONS OF WRN IN RESPONSE TO DNA DOUBLE-STRAND BREAKS
J David, Director And Professor
University Of Texas Sw Med Ctr/dallascity: Dallas country: United States (us)
Grant 5R01CA134991-03 from National Cancer Institute
Abstract: Werner Syndrome (WS) is an autosomal recessive condition characterized by an early onset of age-related symptoms. WS patients also experience an increased risk of rare non-epithelial cancers, especially mesenchymalneoplasms such as sarcomas. Fibroblasts isolated from WS cells senesce prematurely in culture and display increased chromosomal aberrations. WRN, the protein mutated in WS, is unique among the RecQ family proteins possesses on exonuclease activity and 32 to 52 helicase in a single polypeptide. There is accumulating evidence suggesting that WRN contributes to the maintenance of genomic integrity through its involvement in various DNA damage repair pathways and plays a role in telomere maintenance. However, the mechanism by which WRN functions in DNA repair, especially in DNA double-strand break (DSB) repair is still elusive. In vitro and indirect evidence leads to the conclusion that WRN may play a role both in homologous recombination (HR) as well as nonhomologous end joining (NHEJ). Recently, evidence showed that WRN is recruited to DNA damage sites and phosphorylated by PIKK3 kinase family in response to DNA DSB. In this proposal, we will test the hypothesis that WRN is recruited to DNA double-strand breaks (DSB) in vivo, to determine the function of WRN phosphorylation, and its involvement in the process of NHEJ or HR in response to DNA damage. Our specific aims are (1) To determine the mechanism by which WRN is recruited to the sites of DNA double-strand breaks; (2) To test the hypothesis that WRN is phosphorylated by DNA-PK in response to DNA double-strand breaks and the phosphorylation status of WRN modulates its functions at DNA damage sites; and (3) To verify the hypothesis that WRN plays a role in nonhomologous end joining (NHEJ) and/or homologous recombination (HR) pathways of DNA double-strand break repair. Accomplishment of the proposed research would lead to the understanding of WRN´s function in response to DNA damage and help further elucidate the role of WRN in cancer. Werner Syndrome (WS) is an autosomal recessive condition characterized by an early onset of age-related symptoms. WS patients also experience an increased risk of rare non-epithelial cancers, especially mesenchymal neoplasms such as sarcomas. WRN, the protein defective in WS is involved in maintenance of genomic integrity and DNA damage repair. To understand the function of WRN in DNA damage response, we proposed to (i) determine the mechanism by which WRN is recruited to the sites of DNA damage in vivo, (ii) find out kinases that are responsible for WRN phosphorylation and (iii) determine the biological significance of WRN phosphorylation in the process of DNA double strand break repair mediated through non- homologous end joining and/or homologous recombination. The results obtained with these experiments will further elucidate the mechanism by which WRN contributes in genome maintenance, cancer and DNA repair
Keywords: Aberrant Chromosome; Abnormalities, Chromosomal; Affect; age dependent; age related; Alanine; Alanine, L-Isomer; Antimorphic mutation; Aspartic Acid; Assay; base; Bioassay; Biochemical; Biologic Assays; Biological; Biological Assay; Cancers; cell imaging; Cell Line; Cell Lines, Strains; CellLine; Cells; cellular imaging; Chromatin; Chromosomal Aberrations; Chromosomal Alterations; Chromosome Aberrations; Chromosome abnormality; Chromosome Alterations; Chromosome Anomalies; Connective Tissue Sarcoma; cultured cell line; Cytogenetic Aberrations; Cytogenetic Abnormalities; Deoxyribonucleic Acid; DNA; DNA Binding; DNA Binding Interaction; DNA Damage; DNA Damage Repair; DNA Double Strand Break; DNA Helicases; DNA Injury; DNA Repair; DNA unwinding enzyme; DNA Unwinding Proteins; DNA- PKcs protein; DNA-activated protein kinase; DNA-Activated Protein Kinase Catalytic Subunit; DNA-dependent protein kinase; DNA-dependent protein serine-threonine kinase; DNA-PK; DNA-PKcs; DNAPK; DNPK1; Dominant Negative; Dominant-Negative Mutant; Dominant-Negative Mutation; Double Strand Break Repair; early onset; EC 2.7; Electromagnetic, Laser; endo.SceI; endodeoxyribonuclease Sce I; endodeoxyribonuclease SceI; Event; Exonuclease; experience; experiment; experimental research; experimental study; Family; Family member; Fibroblasts; G22P2; Genetic Alteration; Genetic Change; Genetic defect; Genome; genome mutation; Genomics; Health; heavy metal lead; heavy metal Pb; helicase; homologous recombination; Human; human WRN protein; Human, General; Hyper-Radiosensitivity Of Murine SCID Mutation, Complementing 1; HYRC1; I-SceI; In Vitro; in vivo; Interruption; kinase inhibitor; Kinases; Kinetic; Kinetics; Knockout Mice; Ku Antigen, 80-Kd Subunit Gene; Ku80; L-Alanine; L-Aspartic Acid; Laboratories; Lasers; Lead; Life; Maintenance; Maintenances; malignancy; Malignant Neoplasms; Malignant Soft Tissue Neoplasm; Malignant Tumor; Malignant Tumor of the Soft Tissue; Mammals, Mice; Man (Taxonomy); Man, Modern; Measures; Mediating; meganuclease I-SceI; Mesenchymal Cell Neoplasm; Mesenchymal Cell Tumor; Mesenchymal Neoplasm; Mesenchymal Tumor; Mice; Mice, Knock-out; Mice, Knockout; Murine; Mus; mutant; Mutate; Mutation; neoplasm/cancer; NHEJ; Non-Homologous End Joining; Nonhomologous DNA End Joining; Null Mouse; p350; p460 protein; pathway; Pathway interactions; Patients; Pb element; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; polypeptide; PRKDC; Process; Progeria, Adult; Protein Family; Protein Phosphorylation; protein protein interaction; Radiation, Laser; Radiation, X-Rays; Radiation, X-Rays, Gamma-Rays; RECQ3 protein, human; RECQL2 protein, human; recruit; Recruitment Activity; repair; repaired; Reporter; Research; research study; response; Risk; Roentgen Rays; Role; sarcoma; Sarcoma of the Soft Tissue and Bone; Sarcoma, Soft Tissue; SceI endonuclease; SCID protein; Site; social role; spatial relationship; Study Section; Symptoms; System; System, LOINC Axis 4; Telomere Maintenance; Testing; Transphosphorylases; Unscheduled DNA Synthesis; Werner Syndrome; Werner syndrome helicase, human; Werner Syndrome Protein; Werner`s syndrome; WRN Protein; WRN protein, human; X-Radiation; X-Rays; Xrays; XRCC5; XRCC5 gene; XRCC7; XRCC7 protein
Relevance: Werner Syndrome (WS) is an autosomal recessive condition characterized by an early onset of age-related symptoms. WS patients also experience an increased risk of rare non-epithelial cancers, especially mesenchymal neoplasms such as sarcomas. WRN, the protein defective in WS is involved in maintenance of genomic integrity and DNA damage repair. To understand the function of WRN in DNA damage response, we proposed to (i) determine the mechanism by which WRN is recruited to the sites of DNA damage in vivo, (ii) find out kinases that are responsible for WRN phosphorylation and (iii) determine the biological significance of WRN phosphorylation in the process of DNA double strand break repair mediated through non- homologous end joining and/or homologous recombination. The results obtained with these experiments will further elucidate the mechanism by which WRN contributes in genome maintenance, cancer and DNA repair
Project start date: 2009-04-01
Project end date: 2014-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PA-07-070
5R01CA134991-03 (2011): $293198
NOTCH AND STEM CELLS IN LUNG CANCER TREATED WITH ERLOTINIB PLUS NOTCH INHIBITOR.
J David, Professor
University Of Texas Md Anderson Can Ctrcity: Houston country: United States (us)
Grant 1R21CA153017-01A1 from National Cancer Institute
Abstract: We will conduct a preliminary assessment of whether efficacy of erlotinib in NSCLC is reduced if tumors have high Notch pathway and CSC marker expression, and whether addition of the 3-secretase inhibitor RO4929097 will abrogate the negative impact of high Notch/CSC marker expression. We will also assess whether Notch and CSC markers correlate with EMT and EGFR markers, and whether baseline marker signatures predict for efficacy of erlotinib alone or the RO4929097-erlotinib combination. We will assess whether exposure to erlotinib alone enriches tumors for Notch/CSC/EMT markers as a mechanism of acquired resistance, and whether addition of RO4929097 to erlotinib prevents this enrichment or alters impact of erlotinib on EGFR pathway signaling. We will assess impact of host Notch SNPs on tumor Notch/ CSC/ EMT/ EGFR marker expression and on efficacy of erlotinibRO4929097. We may also assess them as a secondary goal. Specifically, where tissue quantity and patient numbers permit, we will conduct preliminary assessments of a) Notch/CSC/EMT marker expression in tumors with vs without other erlotinib resistance factors (eg, T790M mutation, IGF-1R activation, c-Met amplification); b) how Sonic Hedgehog (SHH) and Wnt pathway components modulate or augment the impact of the Notch pathway on EGFR TKI efficacy; c) Notch/CSC/EMT marker expression in EGFR-mutant vs -wild type tumors. Specific Aim 1 We will obtain pre-treatment biopsies from patients receiving erlotinib RO4929097 and will assess Notch pathway, CSC, EMT and EGFR pathway markers by immunohistochemistry (IHC) and reverse phase protein arrays (RPPAs). We will a) correlate expression of Notch and CSC markers with each other and with EMT and EGFR markers; b) correlate marker expression with tumor shrinkage after RO4929097 initiation; c) correlate marker expression with tumor shrinkage after erlotinib initiation; d) compare marker expression in patients with vs without recent erlotinib exposure Specific Aim 2 We will obtain repeat NSCLC biopsies 6 weeks after initiating therapy with erlotinib RO4929097 and will compare change in Notch, CSC, EMT and EGFR marker expression in patients who received erlotinib alone to those who also received RO4929097. Specific Aim 3 We will assess how host Notch pathway SNPs (assessed in peripheral blood mononuclear cells [PBMCs]) correlate with a) tumor expression of Notch, CSC, EMT and EGFR markers; b) change in tumor markers with erlotinib RO4929097; c) efficacy of erlotinib RO4929097. Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer cases. About 10% of NSCLCs have activating mutations of the epidermal growth factor receptor (EGFR) gene, and tumors with these mutations are sensitive to EGFR tyrosine kinase inhibitors (TKIs) such as erlotinib. We are conducting an NCI-sponsored trial in which erlotinib-resistant NSCLC patients have the 3-secretase/ Notch inhibitor RO4929097 added to their erlotinib. While erlotinib is less effective vs EGFR wild type tumors, therapeutic benefit is nevertheless seen
Keywords: Accounting; Biopsy; c-erbB-1 Proto-Oncogenes; Cancer Patient; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Erinaceidae; Erlotinib; Exposure to; Goals; Immunohistochemistry; inhibitor/antagonist; Malignant neoplasm of lung; meetings; mutant; Mutation; Non-Small-Cell Lung Carcinoma; notch protein; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Phase; prevent; Protein Array; public health relevance; R Factors; Resistance; secretase; Signal Pathway; Stem cells; Therapeutic; Tissues; tumor; Tumor Markers
Relevance: Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer cases. About 10% of NSCLCs have activating mutations of the epidermal growth factor receptor (EGFR) gene, and tumors with these mutations are sensitive to EGFR tyrosine kinase inhibitors (TKIs) such as erlotinib. We are conducting an NCI-sponsored trial in which erlotinib-resistant NSCLC patients have the ¿-secretase/ Notch inhibitor RO4929097 added to their erlotinib. While erlotinib is less effective vs EGFR wild type tumors, therapeutic benefit is nevertheless seen
Project start date: 2011-03-02
Project end date: 2013-02-28
Budget start date: 2-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PAR-08-025
1R21CA153017-01A1 (2011): $253472
STEM CELL RESEARCH TRAINING PROGRAM
J David, Professor
University Of Wisconsin Madisoncity: Madison country: United States (us)
Grant 5T32AG027566-05 from National Institute On Aging
Abstract: We propose to establish an interdisciplinary postdoctoral training program for all aspects of research into the biology of stem cells. A major strength of this program is the range of expertise at University of Wisconsin in stem cell biology - from basic embryology, through clinical trials, to ethics and policy research. Postdoctoral trainees from specific areas of developmental biology, neuroscience, endocrinology, blood science, heart science, bioengineering, and other specialties such as policy and ethics will enter this program and be matched with the primary mentor appropriate for their area of interest. In addition, there will be opportunities to study the ethical implications of stem cell research. Key aspects of the program include (i) establishing mentor committees with direct interest in stem cell biology, (ii) encouraging active interactions with other laboratories involved with stem cell research through seminars, journal clubs, workshops, and retreats, (iii) attending UW-Madison courses related to stem cell biology and research ethics, and (iv) interacting with the key institutes on campus which have become central to stem cell biology, including the Biotech Center, Waisman Center, Primate Center, and the non profit organization WiCell. The trainees will participate in a number of newly developed courses in stem cell biology - from basic principles to applications. They will be made aware of the huge commercial implications of stem cell biology and the latest thinking on translating this potential into clinical trials. The investigators aim to train a future generation of principal investigators within this important new field. All laboratories in this program which work with human embryonic stem cells will continue to use only Federally-approved cell lines, specifically WA01, WA07, WA09, WA13, and/or WA14
Keywords: Research Training; Stem Cell Research; Training Programs
Project start date: 2005-05-01
Project end date: 2011-04-30
Budget start date: 1-MAY-2009
Budget end date: 30-APR-2011
5T32AG027566-05 (2009): $139870
GENE TRANSFER FOR PREVENTION OF DIABETIC NEUROPATHY
J David, Professor And Chair
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Grant 5R01NS038850-11 from National Institute Of Neurological Disorders And Stroke
Abstract: We have developed recombinant, replication-incompetent, genomic herpes simplex virus (HSV)-based vectors for gene transfer to the nervous system. In the previous period of funding we have demonstrated that HSV- mediated expression of neurotrophin-3 (NT-3), vascular endothelial growth factor (VEGF), or erythropoietin (EPO) in dorsal root ganglion (DRG) in vivo prevents the progression of neuropathy in mice with streptozotocin (STZ)-induced diabetes. In this competing renewal, we will continue to exploit the natural tropism of HSV vectors for peripheral sensory neurons of the (DRG) to further test the hypothesis that gene transfer of neuroprotective factors to the DRG by means of an HSV-based vector can be used to prevent the progression of diabetic neuropathy, with the ultimate aim of developing a therapy that will be effective for the treatment of human neuropathy. Four specific aims are described. (1) to determine whether HSV-mediated neuroprotective factor expression prevents the loss of DRG neurons in the streptozocin model of type 1 diabetes; (2) to examine the effect of HSV-mediated delivery of neuroprotective factors in the treatment of diabetic neuropathy in the db/db mouse model of type 2 diabetes; (3) to construct a vector with a regulatable switch to control transgene expression safely; and, (4) to test the effect of intermittent regulated protective factor expression on the progression of diabetic neuropathy in the STZ-diabetic and db/db mouse. Neuropathy is an important and often disabling complication of both type 1 and type 2 diabetes mellitus, and there are no currently available treatments that are effective in preventing the progression of the complication. These studies are designed to develop novel agents that could be used to treat the disease effectively
Keywords: Adverse effects; afferent nerve; Afferent Neurons; Age; Analgesics; Animal Model; Animals; Autonomic Dysfunction; autonomic neuropathy; base; behavior measurement; Behavioral; Binding Sites; Biological Assay; C Fiber; Cell Death; Cisplatin; Code; Complication; Complications of Diabetes Mellitus; Data; db/db mouse; Detection; Development; Diabetes Mellitus; diabetic; Diabetic mouse; Diabetic Neuropathies; diabetic patient; Diabetic Polyneuropathies; Disease; DNA; DNA Binding Domain; Dose; effective therapy; Elements; Engineering; Enzyme-Linked Immunosorbent Assay; Erythropoietin; Event; expression vector; Fiber; foot; Functional disorder; Funding; Gene Expression; Gene Transfer; gene transfer vector; Genes; Genetic Models; Genetic Transcription; Genomics; Green Fluorescent Proteins; Health; Heterodimerization; Histologic; Homebound Persons; Homologous Gene; HSV vector; Human; human disease; Hyperglycemia; Immunophilins; Immunosuppressive Agents; In Vitro; in vivo; Injection of therapeutic agent; Insulin-Dependent Diabetes Mellitus; Kinetics; Life; Ligation; Lipids; Measures; Mediating; Mediator of activation protein; Modeling; Monitor; Motor; mouse model; Mus; Nerve; Nerve Fibers; Nerve Growth Factors; nerve supply; Nervous system structure; Neural Conduction; Neuroglia; Neurologic Deficit; Neurons; Neuropathy; neurotrophic factor; Neurotrophin 3; Non-Insulin-Dependent Diabetes Mellitus; novel; NTF3 gene; Outcome Measure; painful neuropathy; Pathogenesis; Peptides; Peripheral; Peripheral Nerves; Peripheral Nervous System; Pharmacologic Substance; phase 1 study; Phosphotransferases; Polyneuropathy; pre-clinical; preclinical study; prevent; Prevention; Promotor (Genetics); protective effect; Proteins; pyridoxine; Rattus; receptor; Recombinants; Research Design; research study; response; Reverse Transcriptase Polymerase Chain Reaction; Reverse Transcription; RNA; Rodent; secondary outcome; Sensory; Simplexvirus; Skin; small molecule; Solutions; Spinal Cord; Spinal Ganglia; Spinal nerve structure; Staining method; Stains; Stimulus; Streptozocin; Streptozocin Diabetes; subcutaneous; Surface; System; Tacrolimus Binding Proteins; Testing; Time; Tissues; Toxic effect; transgene expression; Transgenes; Translations; Tropism; type I and type II diabetes; Vascular Endothelial Growth Factors; vector; Withdrawal; Work
Project start date: 1999-07-23
Project end date: 2013-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-070
5R01NS038850-11 (2011): $325850
THE ROLE OF HIPPOCAMPAL SEQUENCE PLAY IN LEARNING AND DECISION MAKING
J David, Assistant Professor
Johns Hopkins Universitycity: Baltimore country: United States (us)
Grant 5R01MH085823-02 from National Institute Of Mental Health
Abstract: We aim to study how populations of neurons in the hippocampus and prefrontal cortex support navigational learning and decision-making. During awake behavior, hippocampal neurons are active in precise sequences that reflect trajectories taken in the past, and hypothetical trajectories to be taken in the future. This phenomenon is known as awake replay. The broad objective of this proposal is to investigate a possible role for awake replay in learning and decision making. Previous data together with a model suggest a framework in which replay sequences can be used to learn trajectories to reward locations, and also provide the ability to "look ahead" along hypothetical future trajectories during decision-making. We make use of high-density tetrode electrophysiology techniques which allow simultaneous recording of hundreds of neurons in multiple brain regions, in freely behaving animals. We record single unit and wideband EEG activity during the learning and performance of navigational tasks. Our preliminary data show that replay undergoes learning-related changes, that replay in the hippocampus is closely coordinated with activity in prefrontal cortex which is an area strongly associated with decision-making, and that replay at choice points is predictive of navigational decisions. We will pursue these preliminary results in three specific aims. Understanding how replay relates to learning will provide novel insight into offline mechanisms of learning, which are increasingly thought to be important, both in normal cognition, and in diseases such as Alzheimer´s disease, autism and schizophrenia. Understanding how the hippocampus interacts with prefrontal cortex during replay will shed further light on the importance of offline activity, and also provide novel insight into the way that the hippocampus conveys specific information to areas of the brain that influence behavior. Hence, we will investigate a novel and important link between hippocampal neural activity and behavioral outcomes. Finally, understanding how hippocampal replay is organized during decision-making will provide novel insight into hippocampal memory retrieval. Understanding how the hippocampus supports memory is a major goal in medicine, given the involvement of the hippocampus in memory problems in Alzheimer´s disease, Korskoff´s syndrome, normal aging, stroke and temporal lobe epilepsy. Further, this study may reveal novel prefrontal mechanisms of planning and decision making, which may shed light on those brain diseases that have been associated with degraded prefrontal function, such as schizophrenia and autism. Ultimately, the search for the neural basis of memory is a fundamental goal with profound implications for the development of therapeutic targets in brain disease. Despite the prevalence of memory disorders associated with damage to the hippocampus, from such conditions as Alzheimer´s disease, aging, stroke and epilepsy, we lack a mechanistic understanding of how hippocampal neurons encode and retrieve memories, which could help us address these problems. However, recently developed techniques for recording from large numbers of neurons simultaneously in awake, freely behaving animals, have begun to reveal patterns of activity across hippocampal neurons that may provide a model of memory processing. This study will take these results further by asking how the retrieval of specific memories by the hippocampus drives the selection of appropriate behavior, thus shining light on the very processes that may go awry when memory fails
Keywords: Address; Aging; Alzheimer`s Disease; Animals; Area; Autistic Disorder; awake; base; Behavior; behavior influence; Behavioral; Brain; Brain Diseases; Brain region; Cells; Cognition; Data; Decision Making; density; Development; Disease; Electroencephalography; Electrophysiology (science); Epilepsy; Episodic memory; Event; Exhibits; experience; follow-up; Future; Goals; Hippocampus (Brain); insight; Learning; Light; Link; Location; Medial; Medicine; Memory; Memory Disorders; memory process; memory retrieval; Modeling; Neurons; normal aging; novel; Outcome; Pattern; Performance; Play; Population Study; Prefrontal Cortex; Prevalence; Process; prospective; public health relevance; Publications; Rattus; Recruitment Activity; relating to nervous system; Research; research study; response; Retrieval; Rewards; Role; Running; Schizophrenia; Sleep; spatiotemporal; Staging; stroke; Syndrome; Techniques; Technology; Temporal Lobe Epilepsy; Testing; therapeutic target; Time; Training
Relevance: Public Health Relevance Despite the prevalence of memory disorders associated with damage to the hippocampus, from such conditions as Alzheimer´s disease, aging, stroke and epilepsy, we lack a mechanistic understanding of how hippocampal neurons encode and retrieve memories, which could help us address these problems. However, recently developed techniques for recording from large numbers of neurons simultaneously in awake, freely behaving animals, have begun to reveal patterns of activity across hippocampal neurons that may provide a model of memory processing. This study will take these results further by asking how the retrieval of specific memories by the hippocampus drives the selection of appropriate behavior, thus shining light on the very processes that may go awry when memory fails
Project start date: 2010-07-15
Project end date: 2015-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-070
5R01MH085823-02 (2011): $405900
HIGH DENSITY TETRODE RECORDING IN FREELY BEHAVING MOUSE MODELS OF MENTAL DISEASE
J David, Assistant Professor
Johns Hopkins Universitycity: Baltimore country: United States (us)
Grant 5R21MH086702-02 from National Institute Of Mental Health
Abstract: We aim to explore a novel experimental approach to understanding mental disease. Our long-term goal is to understand how schizophrenia affects information processing in the brain. To this end, I will apply high-density tetrode electrophysiology techniques to record in multiple animal models of schizophrenia, during free behavior. Much recent work in schizophrenia has focused on enhanced activity of the default mode network brain regions which are particularly co-active during rest periods, and which include the hippocampus. This converges with recent work by myself and others investigating normal hippocampal function, and particularly, the phenomenon of awake replay, in which large numbers of hippocampal neurons fire in sequences that relate to ongoing task demands. For example, awake replay can "look-ahead" along spatial trajectories ahead of an exploring animal to explore hypothetical outcomes. We propose to investigate neural activity during movement and rest, in two different mouse models of schizophrenia, in order to (1) determine whether there are disruptions to rest period activity in particular, reminiscent of disruption to the default mode network, and (2) determine whether two distinct genetic mouse models of schizophrenia exhibit commonalities, especially with respect to rest period activity. Our preliminary data show that hippocampal neural activity is affected in a forebrain-specific calcineurin knockout mouse model, in a manner specific to rest periods. We are now in a position to determine the effect of these changes on awake replay. We will also consider a transgenic DISC1 mouse model, which exhibits reduced parvalbumin-immunoreactivity, as is also seen in schizophrenia patients. This is particularly interesting because parvalbumin-containing interneurons initiate and sculpt awake replay events in normal animals, and loss of such inhibition might be expected to lead to a similar outcome as observed in the calcineurin mouse. Determining how information processing in rest periods is disrupted is a key goal, which will make possible future studies to determine how disruptions to awake replay affect ongoing task performance, how heterogeneous disease mechanisms may lead to similar behavioral phenotypes, and ultimately how circuit level dysfunction may be treated therapeutically. Schizophrenia is a psychiatric disorder that affects almost 1% of the world population, and represents the seventh most costly medical illness to society. This proposal seeks to explore a novel experimental approach to the disease, looking for commonalities between two genetically distinct animal models, and specifically targeting neural activity at the level of circuits using a cutting edge electrophysiological technique that allows for the simultaneous recording of hundreds of neurons in freely behaving animals. This work will pave the way for a better understanding of how similar circuit level phenotypes might arise out of diverse genetic models, and generate a novel understanding of the neural basis of mental phenomena that might be applied in the future to the diagnosis and treatment of human patients
Keywords: Affect; Animal Model; Animals; Area; awake; base; Behavior; Behavioral; behavioral impairment; Brain; Brain region; C-terminal; Calcineurin; Characteristics; Data; density; Diagnosis; Disease; Disinhibition; Dominant-Negative Mutation; Electroencephalography; Electrophysiology (science); Event; Exhibits; experience; Fire - disasters; follow-up; free behavior; Frequencies (time pattern); Functional disorder; Future; Generations; Genes; Genetic; Genetic Models; Goals; Hippocampus (Brain); Human; immunoreactivity; Impairment; information processing; inhibitory neuron; interest; Interneurons; Knock-out; Knockout Mice; Lead; Learning; Light; Medical; Memory; Mental disorders; Modeling; mouse model; Movement; Mus; neural information processing; Neurons; novel; Outcome; Parvalbumins; Patients; Phenotype; Population; Positioning Attribute; Process; Prosencephalon; Psyche structure; public health relevance; relating to nervous system; Reporting; Research; research study; response; Rest; Schizophrenia; Sleep; Societies; Task Performances; Techniques; Technology; Testing; Transgenic Mice; Transgenic Model; Transgenic Organisms; Work
Project start date: 2010-07-15
Project end date: 2012-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-09-164
5R21MH086702-02 (2011): $147600
SYNAPTIC PROCESSING IN THE BASAL GANGLIA
J David, Professor
University Of Washingtoncity: Seattle country: United States (us)
Grant 5R01MH066128-08 from National Institute Of Mental Health
Abstract: Vocal learning in songbirds is a unique, experimentally accessible model of human vocal learning that also exemplifies the acquisition of complex social behavior. A male songbird learns his courtship song by first memorizing his father´s song, and later using auditory feedback to match his own song to his memory of his father´s song. One major advantage to this model system is the existence of separate forebrain circuits involved in producing the song and in learning it. The pathway needed for learning song involves the basal ganglia, a set of brain regions known in mammals to be important for motor control, motor learning and a variety of cognitive functions. Because of the relatively simple circuitry for song learning, we hypothesize that understanding vocal learning in songbirds will provide general insights into learning mechanisms in mammals, including humans. Specifically, we propose to explore cellular mechanisms underlying information transfer and processing through the learning circuit. The experiments proposed here will use electrophysiological and neuroanatomical approaches to understand the structural, functional and molecular components of the wiring of this pathway. We will (1) determine the cellular specializations underlying an unusually powerful inhibitory synapse in the learning circuit; (2) test whether that "inhibitory" synapse can drive activity in postsynaptic neurons in vivo; (3) determine the dopamine receptors and neuropeptides in key neurons in one basal ganglia structure essential for learning; (4) test for functional connections in a novel anatomically characterized pathway that could provide song-related information to neurons of the dopamine system; and (5) measure when dopamine is released in the learning circuit, as a first test of whether dopamine may play a role in song learning. These experiments will provide necessary fundamental information about how the avian learning circuit accomplishes its normal function. Because of the strong foundation that we and others have built comparing avian song learning circuits with basal ganglia circuits in mammals, the results will yield insights more broadly into how basal ganglia circuits can contribute to learning of complex behavior. They will lay the foundation for higher-level experiments aimed at manipulating information processing in the learning circuit to alter learning in a predictable fashion. Although this work is focused on the basic mechanisms underlying cognitive function, because a variety of disorders such as autism spectrum disorder, schizophrenia, Parkinson disease and Huntington disease involve the basal ganglia, this research also has the potential to have long- term impact on those disorders. PUBLIC HEALTH RELEVANCE These experiments take advantage of the strong parallels that we have demonstrated between songbird basal ganglia circuits and those of mammals. Together, they address cellular mechanisms of information processing in the basal ganglia that underlie learning of a complex social behavior in juveniles and modification of such behavior in adults. The results will guide understanding of the role of the basal ganglia in cognitive processes such as learning and ultimately may shed light on disorders in which the basal ganglia are implicated, such as autism and schizophrenia
Keywords: 6-Cyano-7-nitroquinoxaline-2, 3-dione; Address; Adolescent; Adult; Affect; Aggressive behavior; Agonist; AMPA Receptors; Anterior; Area; Auditory; auditory feedback; autism spectrum disorder; Autistic Disorder; Automobile Driving; Axon; Basal Ganglia; Behavior; Bicuculline; Biological Models; Birds; Brain region; cell body (neuron); Cells; Cognitive; cognitive function; Complex; Courtship; Denervation; Dependence; Disease; Dopamine; Dopamine D1 Receptor; Dopamine D2 Receptor; Dopamine Receptor; dopamine system; Enkephalins; extracellular; Fathers; Foundations; Globus Pallidus; Glutamate Receptor; Goals; Health; Human; Huntington Disease; In Situ Hybridization; In Vitro; in vitro activity; in vivo; information processing; Inhibitory Synapse; insight; Interdisciplinary Study; interest; Label; Learning; Light; male; Mammals; Measures; Mediating; Memory; Messenger RNA; Modeling; Modification; Molecular; Molecular Profiling; Motivation; motor control; motor learning; Nerve; Neural Pathways; Neurons; Neuropeptides; novel; Output; Parkinson Disease; patch clamp; Pathway interactions; Play; Population; postsynaptic; Presynaptic Terminals; Process; Production; Prosencephalon; receptor; Receptor Activation; Research; research study; response; Role; Scanning; Schizophrenia; Signal Transduction; Social Behavior; Songbirds; Speech; Structure; Substance P; Synapses; Synaptic plasticity; Testing; Thalamic Nuclei; Thalamic structure; Ventral Tegmental Area; Vertebrates; vocal learning; Whole-Cell Recordings; Work; zebra finch
Relevance: These experiments take advantage of the strong parallels that we have demonstrated between songbird basal ganglia circuits and those of mammals. Together, they address cellular mechanisms of information processing in the basal ganglia that underlie learning of a complex social behavior in juveniles and modification of such behavior in adults. The results will guide understanding of the role of the basal ganglia in cognitive processes such as learning and ultimately may shed light on disorders in which the basal ganglia are implicated, such as autism and schizophrenia
Project start date: 2002-07-01
Project end date: 2013-02-28
Budget start date: 1-APR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01MH066128-08 (2011): $343787
Sponsored Links Excellgen http://Excellgen.com
OCULAR AND HEALTH TRAJECTORIES: THE SEE STUDY
J David, Associate Professor
University Of Miami School Of Medicinecity: Miami country: United States (us)
Grant 1R21EY021187-01 from National Eye Institute
Abstract: Vision problems are among the top ten conditions responsible for disability among Americans; these conditions are often associated with an increased risk of death. This study will utilize structural equation modeling (SEM) to study linkages between visual impairment and other ocular conditions with morbidity status and mortality risk to answer the following questions 1) What is the relative contribution of ocular system declines to changes in overall health and functional status?; 2) Do improvements in ocular health, either via effective medical management of ocular conditions or risk factor modification (e.g., smoking cessation) lead to improvements in functional status and lower mortality risk?; 3) Do ocular system declines directly lead to an increased risk of mortality, or do these changes also indirectly influence mortality risk via adverse impacts on functional status indicators?; and 4) What is the role of visual impairment (VI) in the aging process? That is, is VI simply a symptom of underlying frailty or is it independently associated with worsening health? Data for this study will come from the Salisbury Eye Evaluation (SEE) Project, a study of vision and function in older adults. The study enrolled a random sample of 2520 community residents 65 to 84 years of age. Participants underwent comprehensive vision testing and ocular examinations, and were administered multiple tests of functional status, portions of which were repeated in three subsequent rounds over a 10-year period. Mortality status was obtained thru 2003; full linkage of the entire cohort with the National Death Index (NDI) thru 2009 is proposed as part of this application and will yield a sufficient number of deaths and person-years of follow-up to reliably detect hazard ratio associations smaller than 1.34 with 80% power. This statistical power, in combination with the comprehensive assessment of ocular and functional status indicators assessed over four time periods, will enable us to 1) examine associations between change in visual acuity levels and change in functional mobility and other morbidity indicators (e.g., grip strength) and depressive symptoms among SEE participants following adjustment for time-varying ocular conditions, and select health conditions and chronic disease risk factors; and 2) examine associations between vision related factors (e.g. changes in visual acuity status, presence of ocular conditions, ocular healthcare utilization) and health-related factors (e.g. health behaviors, health conditions, and mortality risk). Proposed study aims support key goals of the National Eye Institute (NEI) Ocular Epidemiology Strategic Planning Report, including 1) Determine the burden of eye diseases and their visual outcomes in a changing population, and 2) develop new methodologies to support ophthalmic epidemiological research. Vision problems are among the top ten conditions responsible for disability among Americans and are also associated with an increased risk of death. This study will examine, in a comprehensive manner, the role of the development of these vision conditions on health, functioning, and risk of death. It will also determine if improvements in visual functioning due to either treatment (e.g., cataract surgery) and/or improvements in lifestyle (e.g., smoking cessation) leads to improved health and a lower risk of death
Keywords: 0-11 years old; Active Follow-up; Address; advanced age; Age-Years; Aged 65 and Over; Aging; Aging Process; Aging-Related Process; American; Annual Reports; Automobile Driving; Cancers; Caring; Cataract Extraction; cataract surgery; cease smoking; Cessation of life; Cessation of smoking; Child; Child Youth; children; Children (0-21); Chronic Disease; chronic disease/disorder; chronic disorder; Chronic Illness; Cognitive; cohort; Communities; Contrast Sensitivity; Coupled; Data; Data Set; Dataset; Death; depressive; depressive symptoms; Diagnosis; disability; disease risk; disorder risk; driving; Drivings, Automobile; Elderly; Elderly, over 65; elders; Emotional Depression; enroll; Enrollment; Epidemiologic Research; Epidemiologic Studies; Epidemiological Studies; Epidemiology; Epidemiology Research; Equation; Evaluation; Evaluation Studies; experience; Eye; Eye diseases; eye disorder; Eyeball; flexibility; follow-up; frailty; functional status; geriatric; Goals; grasp; Grips; hazard; Health; Health behavior; health care service utilization; Health Care Utilization; health services utilization; Health Status; Health Surveys; healthcare service utilization; healthcare utilization; heavy metal lead; heavy metal Pb; Human, Child; improved; indexing; Individual; late life; later life; Lead; Level of Health; Life; Life Style; Lifestyle; Link; long-term study; Longitudinal Studies; malignancy; Malignant Neoplasms; Malignant Tumor; Medical; Method LOINC Axis 6; Methodology; Methods and Techniques; Methods, Other; Modeling; Modification; Monitor; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Names; National Eye Institute; National Health and Nutrition Examination Survey; National Health Interview Survey; neoplasm/cancer; NHANES; older adult; older person; ophthalmopathy; Outcome; Partial Sight; Participant; Pb element; Persons; Population; Productivity; public health relevance; QOL; Quality of life; Relative; Relative (related person); Reporting; Research; Risk; Risk Factors; Role; Sampling; Scientist; Senescence; senescent; senior citizen; Sight; Smoking; smoking cessation; social role; Social Security Number; SSN; Strategic Planning; Symptoms; Symptoms of depression; System; System, LOINC Axis 4; Techniques; Testing; Time; treatment utilization; Universities; Vision; vision development; Vision Tests; Vision, Diminished; Vision, Low; Vision, Reduced; Vision, Subnormal; Visit; Visual; Visual Acuity; Visual Contrast Sensitivity; visual development; Visual impairment; visual system development; visually impaired; youngster
Relevance: Vision problems are among the top ten conditions responsible for disability among Americans and are also associated with an increased risk of death. This study will examine, in a comprehensive manner, the role of the development of these vision conditions on health, functioning, and risk of death. It will also determine if improvements in visual functioning due to either treatment (e.g., cataract surgery) and/or improvements in lifestyle (e.g., smoking cessation) leads to improved health and a lower risk of death
Project start date: 2011-02-01
Project end date: 2013-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PAR-09-252
1R21EY021187-01 (2011): $243647
MOLECULAR IMAGING AND SYSTEM DYNAMICS OF TUMOR VIROTHERAPY
J David, Associate Professor Of Medicine
Mayo Cliniccity: Rochester country: United States (us)
Grant 1R01CA164241-01 from National Cancer Institute
Abstract: Viruses that specifically replicate and kill tumor cells are highly attractive and novel cancer therapies. After an initial infection, these viruses replicate and spread within the tumor, killing cancer cells while leaving normal cells unharmed. We have developed several viruses that are in clinical trials for patients with incurable tumors. These viruses exhibit different cell killing properties and replication kinetics. Vesicular stomatitis virus (VSV) kills cells rapidly by lysis and spreads from cell to cell by diffusion. In contrast, viruses derived from the Edmonston vaccine strain of measles virus (MV) spread by formation of syncytia. The outcome of tumor therapy with viruses is highly complex since it depends on the dynamic interactions among various populations including tumor cells, the virus, and various cellular components of the immune response and the local architecture of the tumor that may hinder or enable spread of the infection. Understanding the outcomes of such therapy requires a rigorous analysis of the dynamics of therapy in vivo. This necessitates (i) methodologies that enable serial and non-invasive quantification of the tumor cell population and virus load and (ii) mathematical and computational tools to understand these dynamics. We have developed replication competent VSV as well as MV derivatives that express the thyroidal sodium iodide symporter (NIS) in cells. Infected tumor cells express NIS and concentrate radioactive isotopes that can be imaged with single photon emission computerized tomography enabling us to serially and non- invasively monitor the biodistribution and propagation of these virus infected cells in tumors. Simultaneously, we have developed mathematical and computational models that capture the essential components of the dynamic interactions between viruses and their target tumors. In this grant application, we plan to (i) establish the relationship between NIS mediated isotope uptake and intratumoral virus replication in order to be able to determine the absolute population of virus infected cells in the tumor at any time after the start of therapy. This analysis will look at various tumors to accommodate the expected inter tumor variability in NIS expression, isotope concentration as well as amplification and spread of the virus. We will also determine the impact of the initial distribution of virus within the tumor on its spread and the level of tumor control. (ii) We will determine how the architecture of tumors varies across a number of cancer models and attempt to understand how this architecture interacts with the oncolytic virus and influences the therapeutic results. (iii) We will continue to develop more appropriate mathematical and computational models that not only capture the in vivo dynamics but also are able to make meaningful predictions that will be tested in relevant animal models and allow us to optimize therapy. Our in vivo and in silico studies will be statistically correlated using novel approaches to determine the ´simplest´ in silico model that reliably captures the dynamics of therapy and makes meaningful predictions that can be experimentally tested. We hypothesize that understanding the complex dynamics of tumor virotherapy will eventually help us with faster and safer translation of these novel therapies into the clinic and be associated with considerable savings by guiding the design of the relevant animal experiments and clinical trial design
Keywords: 3-Dimensional; Address; Adjuvant; Animal Experiments; Animal Model; anticancer research; Applications Grants; Architecture; Automobile Driving; Autoradiography; base; Biodistribution; cancer cell; Cancer Model; cancer therapy; cell growth; cell killing; Cells; Cellular Structures; Characteristics; Clinic; clinical practice; Clinical Trials; Clinical Trials Design; Collimator; Complex; Computer Simulation; computerized tools; Cytolysis; Data; design; Development; Diffusion; Disease; Drug Delivery Systems; Emission-Computed Tomography; Engineering; Ensure; Environment; Evolution; Exhibits; Giant Cells; Growth; Image; Imaging technology; Immune response; Immune system; Immunohistochemistry; improved; in vivo; Infection; innovation; insight; Isotopes; Killings; Kinetics; Knowledge; Laboratories; Lead; Left; Magic; Malignant Neoplasms; mathematical model; Measles virus; Mediating; Methodology; Modeling; Molecular; molecular imaging; Monitor; neoplastic cell; Normal Cell; novel; novel strategies; Oncogenic Viruses; Oncolytic; Oncolytic viruses; Outcome; particle; Pathway interactions; Patients; Pharmacotherapy; Photons; Population; Population Dynamics; Probability; Property; Radioisotopes; recombinant virus; Relative (related person); Resistance; Resolution; response; Savings; Series; sodium-iodide symporter; success; System; Systems Biology; Technology; Testing; Therapeutic; Therapeutic Agents; therapy outcome; three-dimensional modeling; Time; Translations; tumor; Tumor Burden; uptake; Vaccines; Variant; Vesicular stomatitis Indiana virus; Viral load measurement; Virus; virus characteristic; Virus Diseases; Virus Replication
Relevance: Viruses can be engineered to specifically infect and destroy tumor cells. Efficient spread of the virus in the tumor may lead to its elimination. Taking such viruses from the laboratory to clinical practice requires understanding how the tumor cells and the virus interact and how spreading of infection occurs. This requires the use of technologies that without disturbing the tumor, enable imaging of the distribution of the virus infected cells and how they change in time. Using this data together with advanced computational tools will enable us to understand how the virus and its target cells interact together and how these interactions depend on the local tumor environment that varies across cancer. Such an understanding will hasten the safe and efficient use of these novel therapies to routine clinical practice
Project start date: 2011-09-21
Project end date: 2016-07-31
Budget start date: 21-SEP-2011
Budget end date: 31-JUL-2012
PFA/PA: RFA-CA-11-005
1R01CA164241-01 (2011): $533634
BASIC SCIENCE TRAINING GRANT IN UROLOGY
J David, Assistant Professor
Northwestern Universitycity: Chicago country: United States (us)
Grant 5T32DK062716-07 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Urologic diseases cause significant morbidity, mortality, and burden on the U.S. healthcare system of approximately $11B annually. To meet this healthcare challenge, a new generation of urologic researchers must be trained - both clinical and basic scientists. We propose training these future urologic researchers through the renewed funding of the Urology Training Program at Northwestern University. Our young program has already achieved important milestones, including placing its first trainee into the ranks of Assistant Professor at a distinguished institution. The Department of Urology has a long tradition training thought leaders in clinical and basic urologic research. In this application, we seek funding for two urologic research fellows annually. Fellows will be Ph.D. postdoctoral scientists or M.D., Ph.D. physician-scientists, and we have budgeted accordingly. The Urology Training Program relies upon 16 preceptors from within Urology and other Northwestern departments. Together, these preceptors represent diverse areas of urology, from basic biology and pathogenesis in the urinary tract to outcomes. The Urology Training Program also exploits the many centers, programs, and core facilities of Northwestern to provide a unique training environment that effectively trains fellows for successful careers in clinical and basic urologic research. In addition to the rigorous research components, the program is also structured to facilitate career development with training critical academic skills such as grant writing. An important component of the Urology Training Program is the structure that provides continuous evaluation of trainees, preceptors and the program itself by an internal Executive Committee and an external Steering Committee. Outstanding fellow candidates will be aggressively recruited from renowned Ph.D. programs and from the urology residency program at Northwestern University and its affiliation with the innovative Physician-Scientist Training Program. Only those candidates with a strong desire to pursue academic research and outstanding prior achievement will be considered for training in this highly selective program. Furthermore, every possible effort will be made to enhance diversity by specifically recruiting minority candidates. In summary, we have established a highly successful Urology Training Program. We seek to build upon our early successes by training the next generation of clinical and basic urologic researchers to provide improved diagnostics and therapies for urologic diseases. Urologic diseases afflict millions of Americans annually, costing grave suffering and billions of dollars for treatment. Training is needed to prepare the next generation of urologic researchers. This program will train both clinical and basic scientists for successful careers in urologic research that will then deliver next-generation technologies for the diagnosis and treatment of urologic diseases
Keywords: Basic Science; Grant; Training; Urology
Project start date: 2003-06-01
Project end date: 2015-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-08-226
5T32DK062716-07 (2011): $13504
INDUCTION AND MODULATION OF HOST RESPONSES BY UPEC
J David, Herman L. Kretschmer Professor & Chair
Northwestern Universitycity: Chicago country: United States (us)
Grant 5R01DK042648-19 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Urinary tract infections (UTIs) afflict approximately 7 million women in the U.S. annually, causing significant morbidity and costing $3.5 billion for treatment. Most community-acquired UTIs are due to infection by uropathogenic E. coli (UPEC) that elicit an inflammatory response in the bladder during acute cystitis. Identifying novel therapeutic targets against UPEC is critical because of both the high incidence of UTIs and the increasing prevalence of antimicrobial resistance among UPEC isolates. NFB is a pivotal mediator of inflammation and other innate responses, so we previously examined urothelial NFB responses to UPEC and demonstrated that UPEC possess NFB suppressive activity (NSA). To characterize the molecular basis of NSA, we performed a genetic screen and identified NSA mutants in UPEC that enhance urothelial inflammation, increase apoptosis, and exhibited attenuated intracellular proliferation. The NSA mutants are defective for activities of normal surface structure biosynthesis, and the involved surface structures are potential activators of innate host responses through Toll-like receptors (TLRs). TLR signaling includes the IB kinase complex (IKK), where other receptor signals converge with TLR signals that activate NFB, such as signals induced by EGF and TNF. Since UPEC also modulates EGF and TNF signaling, we hypothesize that UPEC modulation of innate responses by NSA is mediated by bacterial macromolecules that differentially induce TLR signals impinging upon IKK, thus altering key events of UTI pathogenesis that can be exploited to develop novel therapies. Indeed, NSA mutants modulate IB phosphorylation, induce elevated urothelial apoptosis, are defective for intracellular proliferation, and stimulate enhanced protection in a UTI vaccine assay. In Aim 1, we will identify the UPEC molecules(s) mediating NSA. In Aim 2, we will identify the urothelial targets of UPEC NSA. In Aim 3, we will quantify the impact of NSA on UPEC-induced apoptosis and UPEC invasion and intracellular proliferation. In Aim 4, we will determine the therapeutic potential of UPEC NSA mutants as live vaccines. For all aims, we will employ UPEC strain NU14 and isogenic mutants that differentially induce host responses, and we will use both human cell culture and murine UTI models. These studies lay the groundwork for development and optimization of novel and effective live attenuated vaccines for UTI. Urinary tract infections afflict approximately 7 million women in the U.S. annually. The bacteria that cause the majority of infections, uropathogenic Escherichia coli (UPEC), modify the responses of bladder tissues to promote successful infection. This project will define the mechanism that UPEC uses to modify the host responses and thus provides the groundwork for development and optimization of novel, live-attenuated vaccines against urinary tract infection
Keywords: Acute; Acute Cystitis; adaptive immunity; Anabolism; Antigen Presentation; Antimicrobial Resistance; Apoptosis; Attenuated; Attenuated Live Virus Vaccine; Attenuated Vaccines; Bacteria; base; Biological Assay; Bladder; Bladder Tissue; Cell Culture Techniques; chemokine; community-acquired UTI; Complex; cost; cytokine; Development; Epidermal Growth Factor; Epidermal Growth Factor Receptor; Event; Exhibits; Flagellin; Foundations; gene interaction; Genetic Screening; Human; Immune response; Immunity; immunogenic; Incidence; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory Response; innovation; Kidney; Life; Life Cycle Stages; Ligands; Lipopolysaccharide Biosynthesis Pathway; Lipopolysaccharides; macromolecule; Mediating; Modeling; Molecular; Morbidity - disease rate; Mus; mutant; Mutation; new therapeutic target; novel; Pathogenesis; Peptidoglycan; Phosphorylation; Phosphotransferases; Prevalence; Production; public health relevance; Receptor Signaling; response; Role; SECTM1 gene; Signal Pathway; Signal Transduction; Structure; Suppressor Genes; Surface; Testing; Therapeutic; toll-like receptor 4; Toll-like receptors; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Urinary tract infection; Uropathogenic E. coli; Vaccines; Woman
Relevance: Urinary tract infections afflict approximately 7 million women in the U.S. annually. The bacteria that cause the majority of infections, uropathogenic Escherichia coli (UPEC), modify the responses of bladder tissues to promote successful infection. This project will define the mechanism that UPEC uses to modify the host responses and thus provides the groundwork for development and optimization of novel, live-attenuated vaccines against urinary tract infection
Project start date: 1990-04-01
Project end date: 2014-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-07-070
5R01DK042648-19 (2011): $282441
MOUSE BRAIN MODELS OF TUBEROUS SCLEROSIS
J David
Massachusetts General Hospitalcity: Boston country: United States (us)
Abstract: Tuberous sclerosis (TSC) is an autosomal dominant tumor suppressor gene syndrome, characterized by development of distinctive benign tumors (hamartomas) and malformations (hamartias) in multiple organ systems. The brain lesions (cortical tubers, subependymal nodules, and cerebral white matter migration tracts) account for the most severe manifestations in most patients, including seizures, mental retardation, and autism and other developmental disorders. The long-term objective of this project is to develop and analyze authentic brain models ofTSC in the mouse, so that we can better understand the pathogenesis of TSC in the brain and develop effective therapeutic approaches. We have created a floxed Tsc1 conditional allele, which is a critical reagent for this study. In the first aim, we will complete an analysis of mice in which we have engineered complete loss of Tsc1 in the majority of cortical and hippocampal neurons. These mice are severely affected with both spontaneous and inducible seizure activity, a neurologic wasting syndrome, with ectopic and dysplastic neurons in both the cortex and hippocampus, and a myelination defect; all without genetic modification of astrocytes or astrogliosis. This model displays a marked therapeutic response to rapamycin/RADOOl treatment with increase in median survival from 35d to > 100d. We will examine cortical layer organization through use of specific antibodies and BrdU birthdating, assess neurotransmitter-receptor and transcription factor expression alterations, and assess the response to interrupted rapamycin/RADOOl treatment in this model. In the second aim, we will explore a second brain model of TSC, using a regulable neural progenitor cell specific promoter in the form of the rtTA-tet-on system. Detailed studies of cortical development, seizure predilection, and response to rapamycin/RADOOl will be performed. In the third aim, we will explore the specificity of a set of novel brain enhancers for the cortex, and select one or more of them for use in generating a novel .inducible brain model of TSC, using the ere recombinase-estrogen receptor (Cre-ER) system. Through these approaches, we will be able to modulate the amount of recombination in neural progenitor cells, leading to variable fractions of cells lacking Tsc1 during various stages of brain development
Keywords: Accounting; Actins; Affect; Alleles; Antibodies; Architecture; Astrocytes; astrogliosis; Autistic Disorder; Benign; body system; Brain; Bromodeoxyuridine; Cell Fraction; cell type; Cerebrum; Cessation of life; Clinical; Defect; Development; developmental disease/disorder; Disease; Dose; Doxycycline; Dysplasia; Electroencephalography; Engineering; Enhancers; Estrogen Receptors; Event; Functional disorder; Generations; Genes; Genetic; Genetic Recombination; Genetic Transcription; Giant Cells; Goals; Hamartoma; Hippocampus (Brain); human TSC1 protein; human TSC2 protein; improved; in vivo; Inherited; Laboratories; Lead; Lesion; malformation; Mental Retardation; migration; Modeling; Modification; Molecular Genetics; Mus; Mutation; myelination; nerve stem cell; nervous system disorder; nestin protein; Neuroepithelial; Neurologic; neuron loss; Neurons; Neurotransmitter Receptor; Neurotransmitters; Nodule; novel; Pathogenesis; Patients; precursor cell; Predisposition; Process; Promotor (Genetics); Proteins; Proteome; Reagent; receptor expression; recombinase; Regulation; response; Seizures; Signal Pathway; Signal Transduction; Sirolimus; Specificity; Staging; Subependymal; Synapses; Synapsin I; Synapsins; Syndrome; System; Tetanus Helper Peptide; Therapeutic; Time; Tissues; transcription factor; treatment effect; TSC1 gene; TSC2 gene; Tuberous Sclerosis; tumor; Tumor Suppressor Genes; Wasting Syndrome; white matter; Work
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5P01NS024279-23_0015 (2011): $308428
TUBEROUS SCLEROSIS-PATHWAY AND PATHOGENESIS
J David
Brigham And Women´s Hospitalcity: Boston country: United States (us)
Abstract: Tuberous sclerosis (TSC) is an autosomal dominant tumor suppressor gene syndrome, characterized by development of distinctive benign tumors (hamartomas) and malformations (hamartias) in multiple organ systems, due to mutations in either TSC1 or TSC2. The long-term objectives of this project are to understand in detail the functions of the TSC1 and TSC2 genes and proteins, their interacting partners, how they are regulated, and how their loss leads to deregulated growth control in the tumors that develop in TSC patients. The insight achieved will have relevance to growth regulatory mechanisms in general, including in malignant cells. There are four specific aims in this project. First, the importance of phosphorylation at multiple sites on TSC2 by the Akt kinase will be critically tested in vivo through generation and analysis of an allele in the mouse in which these sites are mutated to Ala. Second, we will examine the mechanism and importance in growth control of feedback circuitry in the TSC1/TSC2 signaling pathway that serves to attenuate activation of the Akt kinase when TSC1/TSC2 are lacking, in both cell lines and tumors. Third, we will explore the estrogen responsive growth that is seen in TSC1/TSC2 null cell lines, examining its mechanism, effects on ERa expression and phosphorylation, consequences in terms of gene transcription, and in vivo effects in tumor growth in mice. Fourth, we will examine whether Rheb is the sole downstream effector that is perturbed when TSC1/TSC2 are lacking from cells, whether Rheb activation has the identical effects in cells as the loss of TSC1/TSC2, and identify binding partners of Rheb for downstream signaling effects
Keywords: 9q34; Acute; Affect; Affinity Chromatography; Alanine; Alleles; Attenuated; attenuation; Behavior; Benign; Binding (Molecular Function); Binding Proteins; body system; cancer cell; Cell Culture Techniques; cell growth; Cell Line; Cell Proliferation; Cells; Clinical; Combined Modality Therapy; Complex; Data; Development; Disease; Disease model; Down-Regulation; Drosophila genus; Estrogens; Event; Exhibits; Family member; Feedback; Fibroblasts; Gene Expression; Gene Proteins; Generations; Genes; Genetic Transcription; Genomics; Growth; Growth Factor; Guanosine Triphosphate Phosphohydrolases; Hamartoma; Hepatic Hemangioma; human FRAP1 protein; human TSC1 protein; human TSC2 protein; Impairment; in vivo; inhibitor/antagonist; insertion/deletion mutation; insight; Knock-in Mouse; Knock-out; Laboratories; Lead; Loss of Heterozygosity; malformation; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Mediating; Molecular; mouse model; mTOR Inhibitor; multicatalytic endopeptidase complex; Mus; mutant; Mutate; Mutation; neoplastic cell; Neurons; Null Lymphocytes; Organism; overexpression; Pathogenesis; Pathway interactions; Patients; PDGFRB gene; Phosphorylation; Phosphorylation Site; Phosphotransferases; Protein Binding; protein function; Protein Isoforms; Proteins; PTEN gene; Reagent; Regulation; Reporting; research study; response; restoration; Role; Signal Pathway; Signal Transduction; Sirolimus; Site; Stable Isotope Labeling; Stimulus; Syndrome; Testing; Time; Tissues; Translations; TSC1 gene; TSC1/2 gene; TSC2 gene; Tuberous Sclerosis; tumor; Tumor Cell Line; tumor growth; Tumor Suppressor Genes; Tumor Suppressor Proteins; tumorigenesis; Up-Regulation (Physiology); Work; Yeasts
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5P01CA120964-05_0001 (2011): $435768
THROMBOTIC/FIBRINOLYTIC BALANCE IN CARDIAC TRANSPLANT VASCULOPATHY
J David
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Abstract: Transplantation-associated coronary artery disease (TCAD), characterized by progressive neointimal proliferation and luminal obliteration, remains the major impediment to the long-term survival of heart transplant recipients. Clinical studies suggest that inherited thrombophilias, graft vascular thrombosis, and hyperlipidemia contribute to TCAD. Preliminary data show that cardiac preservation or tissue hypoxia elicits secretion of von Willebrand factor (vWF) and expression of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1), the latter driven by three independent transcriptional motifs in the PAI-1 promoter. Furthermore, in a heterotopic murine vascularized cardiac allotransplant model, TCAD quantified by histomorphometry is significantly reduced in PAI-1 deficient allografts, but exacerbated in tissue plasminogen activator (tPA) null grafts. These data lead us to hypothesize that creation of a fibrin stroma by ischemiadriven thrombosis or inhibited fibrinolysis in the context of ongoing vascular injury, hyperlipidemia, and inflammation, creates a rich matrix driving the development of an occluding neointima. Project 3 for this PPG will elucidate the molecular mechanisms driving thrombus accrual in cardiac allografts. and their role in TCAD pathoqenesis. Aim 1 will elucidate the contribution of thrombosis to TCAD using thrombophilic factor V Leiden mice, mice with a hypomorphic TF mutation, and those with graded levels of vWF. Aim 2 will elucidate the contribution of fibrinolysis to TCAD using mice deficient in PAI-1, tPA, or uPA, as well as mice null for the master switch transcription factor (Egr-1) underlying hypoxic induction of PAI-1. Pharmacological PAI-1 inhibitors will be tested to establish a potential therapeutic target for TCAD where now exist. Aim 3 will determine the role of background hyperlipidemia as a synergistic mechanism driving thrombosis and TCAD, using apolipoprotein E-deficient hypercholesterolemic mice on thrombophilic or thromboresistant backgrounds. This project will interact with Project 1 in areas of PAI-1 biology and hyperlipidemia, Project 2 in areas of thrombosis, hypofibrinolysis, and hyperlipidemia as triggers for venous vascular injury, Project 4 in areas of new anti-thrombotic target development and genetic modifiers of thrombosis, as well as the three Cores. These studies will provide new insights into powerful mechanisms which obliterate vessels in transplanted hearts, and identify new therapies to keep these critical vessels open
Keywords: Affinity; Alloantigen; Allografting; allotransplant; Alteplase; Apolipoprotein E; Area; Atherosclerosis; Automobile Driving; Biological Preservation; Biology; Blood Platelets; Blood Vessels; Cardiac; CCAAT-Enhancer-Binding Proteins; Cells; cellular development; Clinical Research; Coagulation Process; Coronary; Coronary Arteriosclerosis; Coronary Thrombosis; Crossbreeding; Data; Deposition; Development; Elements; Engineering; Equilibrium; Exhibits; factor V Leiden; Fibrin; Fibrinolysis; Functional disorder; Generations; Genes; Genetic; Growth; Heart; heart allograft; heart preservation; Heart Transplantation; Homologous Gene; Human; human data; Hyperlipidemia; Hypoxia; Hypoxia Inducible Factor; Immune; In Situ; in vivo; Incidence; Indium; Inflammation; Inflammatory; Inherited; inhibitor/antagonist; injured; Injury; insight; Ischemia; Isogenic transplantation; Knockout Mice; Lead; Mediator of activation protein; Modeling; Molecular; monocyte; mouse model; Mus; mutant; Mutant Strains Mice; Mutation; novel; Organ; Pathogenesis; Patients; Plasminogen Activator Inhibitor 1; pre-clinical; Process; Promotor (Genetics); research study; response; Rodent; Role; Severities; Signal Transduction; Site; System; Techniques; Testing; therapeutic target; Thrombin; thrombolysis; Thromboplastin; Thrombosis; Thrombus; Tissues; tool; transcription factor; Translating; Transplant Recipients; Transplantation; Vascular blood supply; Vascular Diseases; Vascular Graft; Venous; von Willebrand Factor; Wild Type Mouse
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5P01HL089407-04_5651 (2011): $236153
NEXTGEN DNA BINDING PROTEINS BASED ON TAL DOMAINS
J David, Associate Professor
University Of California Daviscity: Davis country: United States (us)
Grant 1R01GM097073-01 from National Institute Of General Medical Sciences
Abstract: Transcription activator-like (TAL) effector domains were recently discovered to be DNA binding domains. Based on the reported DNA recognition features of the these domains, long arrays of repeats that each recognize one base pair using only two variable amino acids, the structure of this DNA binding domain is unlike any that have been previously described. As such, the protein fold and structural features responsible for specific DNA recognition are highly novel and worthy of elucidation for their own merits. However, the seeming simple recognition code and apparent recognition of a wide spectrum of sequences has important implications as a scaffold for engineering new DNA binding proteins. TAL domains appear to be even more flexible in their recognition properties than zinc fingers (ZFs), the engineering of which have had transformative impact in the areas of gene regulation, genome engineering, genetically modified organisms, and gene therapy. Despite their successes, the difficultly of engineering high-quality ZFs represents a significant bottleneck to their widespread application. We hypothesize that TAL domains will have superior performance characteristics compared to the gold-standard ZF domains for engineering novel DNA- binding proteins. We will test our hypothesis using a combined computational and biochemical approach to examine the protein fold and repeat assembly (Aim 1), elucidate the mechanism and extent of DNA recognition (Aim 2), and investigate the potential of TAL domains for the creation of sequence-specific tools for gene regulation and genome engineering (Aim 3). If successful, this study will provide insights into the structure and function of a novel DNA-binding domain, and an understanding of how those insights can be applied to create tools for genetic modification that would be more broadly accessible and of greater general utility than current methods. KEY WORDS Protein-nucleic acid interactions, engineered zinc fingers, ab initio modeling, protein structure, protein folding, structure-function relationship, genome engineering, gene therapy, computational design. Over the past two decades, proteins have been engineered to regulate and make precise changes to the DNA of living cells, leading to transformative advances in our ability to study and treat human diseases. These methods are based on the ability to reprogram the DNA binding specificity of zinc finger proteins, which is difficult and consequently has restricted their use. Here we will explore the newly discovered TAL DNA binding motif that seems to overcome these limitations, which should enable greater access to more powerful tools for medical research and therapy
Keywords: Affinity; Amino Acids; Area; base; Base Pairing; Binding (Molecular Function); Biochemical; Cells; Characteristics; Code; Computer Simulation; Computer-Assisted Image Analysis; design; DNA; DNA Binding; DNA Binding Domain; DNA-Binding Proteins; Elements; Engineered Gene; Engineering; flexibility; Gene Expression Regulation; gene therapy; Genetic; Genetically Modified Organisms; Genome engineering; Gold; human disease; insight; Knowledge; Life; Mammalian Cell; Medical Research; Methods; Modeling; Modification; novel; nuclease; Nucleic Acids; Performance; Property; protein folding; protein structure; Proteins; public health relevance; Reporting; research study; scaffold; Specificity; Structure; Structure-Activity Relationship; success; Testing; To specify; tool; Transcription Coactivator; transcription factor; Zinc Fingers
Relevance: RELEVANCE Over the past two decades, proteins have been engineered to regulate and make precise changes to the DNA of living cells, leading to transformative advances in our ability to study and treat human diseases. These methods are based on the ability to reprogram the DNA binding specificity of zinc finger proteins, which is difficult and consequently has restricted their use. Here we will explore the newly discovered TAL DNA binding motif that seems to overcome these limitations, which should enable greater access to more powerful tools for medical research and therapy
Project start date: 2011-04-01
Project end date: 2015-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-10-067
1R01GM097073-01 (2011): $269102
GENETIC DISSECTION OF THE TOXOPLASMA GONDII RHOPTRY KINOME
J David, Professor
Dartmouth Collegecity: Hanover country: United States (us)
Grant 1R21AI097018-01 from National Institute Of Allergy And Infectious Diseases
Abstract: Toxoplasma gondii is an important opportunistic infection of AIDS patients. Improved strategies and approaches are urgently needed to more effectively prevent and treat recurrent infections in AIDS. Following primary infection, a chronic life-long T. gondii infection characterized by the presence of persisting cysts containing slowly or nonreplicating bradyzoite parasite forms is established in CNS/brain. Reactivated infection due to cyst rupture during immune suppression causes a difficult to treat and life-threatening Toxoplasmic encephalitis in AIDS. Currently, no vaccine is approved for use in humans to prevent infection and drug treatments for acute infection are suboptimal. Eradicating pre-existing cysts and chronic infection is an excellent approach to control/prevent infection in AIDS. However, no current treatment is effective at eradicating pre-existing cysts and chronic infection. Lack of progress in targeting cyst stages can be attributed to major gaps in our knowledge of parasite biology underlying cyst development and maintenance of chronic infection. Recent progress in understanding the complex host-parasite interaction has revealed that many rhoptry bulb (ROP) proteins appear to be central players in host cell manipulation due to their secretion into the host cell at invasion and their subsequent localization to host cell cytosol/nucleus or their intimate association with the parasitophorous vacuole. Host cell manipulation by these secreted ROP proteins is likely to be critical to the success of parasite biology necessary for acute and chronic infection in vivo. The parasitophorous vacuole appears to be critical for replication of tachyzoites as well as being central to the development of the cyst wall and tissue cyst that characterize chronic infection. We hypothesize that a family of ~ 34 secreted ROP proteins possessing homology to kinases (the "ROP kinome") are outstanding candidates as potential targets to disrupt virulence, cyst development, or chronic infection. Here, we propose to delete each predicted gene member of the secreted ROP kinome in type II T. gondii, and to then ascertain any defect in acute virulence, in early cyst development, or in chronic infection. Using a directed functional genetic approach, this work will identify specific members of the secreted ROP kinome that participate in critical functions required for establishing or maintaining chronic infection in vivo. This project will identify and validate specific secreted ROP kinases as key targets for preventing acute or chronic infection. This project will also develop essential tools and methods required for the future undertaking of larger scale higher throughput knockout and functional genomic projects in type II T. gondii. Consequently, this innovative project has high overall impact by advancing genetic models for dissection of type II biology, by revealing new aspects of host-parasite interaction, and by exposing new targets to prevent or eradicate chronic infection. This project will identify specialized parasite proteins that are essential for the ability of Toxoplasma gondii to establish and maintain a chronic infection. No vaccine is currently available to prevent infection and no drug treatment is available that can eradicate chronic infection. By functional analysis of a family of specialized parasite proteins we expect to identify new targets to prevent or eradicate chronic infection. The information gained in this project helps to uncovering fundamental knowledge of how a parasite manipulates mammalian cells and the mammalian host, and will thus uncover new approaches to eradicate a significant parasite of humans
Keywords: Acquired Immunodeficiency Syndrome; Acute; AIDS-Related Opportunistic Infections; Alleles; Biological; Biological Assay; Biological Process; Biology; Brain; Cell Nucleus; Cells; Chronic; Complex; Cyst; Cytosol; Data; Defect; Development; Dissection; Encephalitis; Exhibits; Failure (biologic function); Family; Family member; Firefly Luciferases; functional genomics; Future; Genes; Genetic; Genetic Models; Human; Image; improved; In Vitro; in vivo; Infection; Infection prevention; innovation; Killings; Knock-out; knockout gene; Knowledge; Life; Maintenance; Mammalian Cell; member; Methods; Modeling; Mus; mutant; Natural immunosuppression; novel strategies; Parasites; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; prevent; Promotor (Genetics); Proteins; Publishing; Recurrence; Reporter; Research; Research Design; Resistance; rhoptry; Rupture; Specific qualifier value; Staging; success; Sum; Tissues; tool; Toxoplasma gondii; Toxoplasmosis; Transgenic Organisms; Vaccines; Vacuole; Validation; Virulence; Work; XRCC5 gene
Relevance: This project will identify specialized parasite proteins that are essential for the ability of Toxoplasma gondii to establish and maintain a chronic infection. No vaccine is currently available to prevent infection and no drug treatment is available that can eradicate chronic infection. By functional analysis of a family of specialized parasite proteins we expect to identify new targets to prevent or eradicate chronic infection. The information gained in this project helps to uncovering fundamental knowledge of how a parasite manipulates mammalian cells and the mammalian host, and will thus uncover new approaches to eradicate a significant parasite of humans
Project start date: 2011-07-01
Project end date: 2013-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-10-069
1R21AI097018-01 (2011): $197500
REGULATED VIRULENCE AND CYST MATURATION IN TOXOPLASMA GONDII
J David, Professor
Dartmouth Collegecity: Hanover country: United States (us)
Grant 5R21AI075931-02 from National Institute Of Allergy And Infectious Diseases
Abstract: Toxoplasma gondii is an important opportunistic infection of AIDS patients. Improved strategies and approaches are urgently needed to more effectively prevent and treat recurrent infections in AIDS. Due to the ease of in vitro culture and cloning, efficiency of transformation, abundance of established genetic tools, and availability of robust murine models of infection, Toxoplasma gondii is increasingly recognized as a model apicomplexan parasite for understanding biology of intracellular parasitism and host immune response. Our laboratory has developed a highly attenuated uracil auxotroph (cps1-1) mutant of T. gondii (RH background) that is proving to be a valuable model organism for understanding critical host-parasite interactions. The cps1-1 mutant invades host cells, but does not replicate in vitro or in vivo in the absence of uracil. Tachyzoites of the uracil auxotroph cps1-1 can be cultured in vitro and following intraperitoneal (i.p.) inoculation into mice elicit potent Th1 responses that confer effective long-term protection from a lethal challenge infection from virulent type I strain RH. Our recent results demonstrate that cps1-1 administered i.p. elicits surprisingly effective long-term protective immunity from lethal primary and chronic infections after peroral challenge with type II ME49 cysts. Most commonly, the natural route of exposure and transmission of Toxoplasma to humans is via oral ingestion of tissue cysts. Bradyzoites released from cysts launch a primary infection in gut mucosa that ultimately leads to the establishment of chronic brain cysts that can later cause Toxoplasmic encephalitis in AIDS. The early host-parasite interactions during T. gondii infection in the gut mucosa are likely to be critical in eliciting innate and adaptive immune responses that lead to immune protection. We propose to develop tet regulated uracil auxotrophy in type I and type II strains with a goal to develop live-attenuated cyst-forming mutants that can be delivered by the natural peroral route. The strains developed in this project should provide valuable tools necessary for the dissection of key host- parasite interactions in early infection in gut mucosa and will also serve as innovative models for studying host response, parasite development, trafficking, cyst biology, immunity, and vaccines. Relevant to Toxoplasmic encephalitis infections in AIDS, the model organisms developed in this project are likely to provide invaluable tools for experimentally addressing cyst formation, cyst maintenance, and cyst recrudescence. This research project will develop and validate model organism strains of Toxoplasma gondii with the property of regulated virulence and development. These model organisms are expected to provide significant improvements in the ability to perform critical investigations into host response, parasite development, and host-parasite biology as these areas relate to new concepts for effective vaccination to prevent or treat brain cysts. Consequently, this research project will accelerate the discovery of new vaccines and treatments for devastating opportunistic parasite infections in AIDS
Keywords: 2, 4(1H, 3H)-pyrimidinedione; 2, 4-dioxopyrimidine; 2, 4-pyrimidinediol; 2-hydroxy-4-(3H)-pyrimidione; 4-hydroxy-2-(1H)-pyrimidione; Acquired Immune Deficiency; Acquired Immune Deficiency Syndrome; Acquired Immuno-Deficiency Syndrome; Acquired Immunodeficiency Syndrome; Acute; Address; AIDS; AIDS Associated Opportunistic Infection; AIDS, Opportunistic Infection; AIDS-Related Opportunistic Infections; Animal Model; Animal Models and Related Studies; Area; Attenuated; auxotrophy; Biology; Body Tissues; Brain; Cells; Chronic; Cloning; Cyst; Development; DISSEC; Dissection; Encephalitis; Encephalon; Encephalons; Engineering; Engineerings; Exhibits; Generalized Growth; Genetic; Genome; Goals; Growth; heavy metal lead; heavy metal Pb; host response; Human; Human, General; Immune; Immune response; Immunity; Immunologic Deficiency Syndrome, Acquired; immunoresponse; improved; In Vitro; in vivo; Infection; Inflammation, Brain; Ingestion; innovate; innovation; innovative; intracellular parasitism; intraperitoneal; Invaded; Investigation; Laboratories; Lead; Life; Maintenance; Maintenances; Mammals, Mice; Man (Taxonomy); Man, Modern; Mice; model organism; Modeling; Mucosa; Mucosal Tissue; Mucous Membrane; Murine; Mus; mutant; Nervous System, Brain; new vaccines; next generation vaccines; novel vaccines; ontogeny; Opportunistic Infections, HIV-Related; Oral; Organism Strains; Parasites; Pathogenesis; Patients; Pb element; prevent; preventing; Property; Property, LOINC Axis 2; R01 Mechanism; R01 Program; Recrudescences; Recurrence; Recurrent; Regulation; Research Grants; Research Project Grants; Research Projects; Research Projects, R-Series; Resistance; resistant; response; Route; RPG; Study models; T. gondii; T. gondii infection; Tetracycline Antibiotic; Tetracyclines; Tissue Growth; Tissues; tool; Toxoplasma; Toxoplasma gondii; Toxoplasma gondii Infection; Toxoplasmosis; trafficking; Transmission; transmission process; Uracil; Vaccination; Vaccines; Virulence; Virulent
Project start date: 2008-04-01
Project end date: 2011-03-31
Budget start date: 1-APR-2009
Budget end date: 31-MAR-2011
PFA/PA: PA-06-181
5R21AI075931-02 (2009): $239850
Sponsored Links Excellgen http://Excellgen.com
FUNCTIONAL ANALYSIS OF CD8 T CELL IMMUNE CONTROL IN TOXOPLASMA GONDII INFECTION
J David, Professor
Dartmouth Collegecity: Hanover country: United States (us)
Grant 5R21AI091461-02 from National Institute Of Allergy And Infectious Diseases
Abstract: Toxoplasma gondii is an important opportunistic infection of AIDS patients. Improved strategies and approaches are urgently needed to more effectively prevent and treat recurrent infections in AIDS. Despite extensive knowledge of host immune response to T. gondii infection, no vaccine is yet approved for use in humans for this parasite or any other protozoan parasite causing opportunistic infection in AIDS. Following an acute primary infection, a chronic life-long T. gondii infection is established in CNS/brain and this chronic infection is characterized by the persisting presence of cysts containing slowly replicating bradyzoite parasite forms. Reactivation of active infection due to rupture of brain cysts in chronic infection is the major mechanism causing difficult to treat and life-threatening Toxoplasmic encephalitis in AIDS. A major gap exists in our understanding of the antigen-specific immune factors that control acute infection and chronic infection. Our preliminary data shows that CD8+ T cells control acute infection and prevent cyst rupture in chronic infection. CD8+ T cells and IFN-3 production are critical mediators of immune control in acute primary and chronic T. gondii infection. We hypothesize that immune control of acute and chronic infection is highly dependent on antigen-specific CD8+ T cell populations that recognize specific parasite-derived epitopes presented to MHC class I during infection. This hypothesis will be tested by investigating the functional importance of defined CD8+ T cell epitopes in mediating control of acute and chronic infection in experiments using mutant virulent strains as well as mutant attenuated vaccine strains that lack known endogenous parasite-derived CD8+ T cell epitope(s). The functional importance of defined CD8+ T cell epitopes in eliciting protective immunity to lethal acute infection will be determined, as well as the ability of a normally pre-established vaccine-induced immunity to protect against a virulent mutant strain lacking specific parasite-derived CD8+ T cell epitope(s). This innovative and exploratory study will functionally reveal whether any of the four recently identified and potentially immunodominant parasite-derived CD8+ T cell epitopes and their corresponding CD8+ T cell populations determine the immune control of acute and chronic T. gondii infection. This project offers significant impact as it is relevant to the understanding of immune control of acute and chronic infection, which will allow future studies to focus on the most critical parasite antigens and their antigen-specific CD8+ T cell responses. This project is also is relevant for the improvement of vaccine design and to identify new interventions to address chronic infection in AIDS. This project is relevant to understanding immune control of acute and chronic infection in the AIDS opportunistic pathogen Toxoplasma gondii. No vaccine is currently available to prevent infection, and no treatment is available to eradicate chronic infection in infected individuals. By addressing fundamental and functional aspects of immune control in T. gondii infection in this project, we expect to reveal new information that can be directly applied to vaccine development to prevent infection, as well as to identify potential interventions to eliminate chronic infection in already infected people who are particularly at risk of serious recurrent infection(s)
Keywords: Acquired Immunodeficiency Syndrome; Acute; Address; AIDS-Related Opportunistic Infections; Antigens; Attenuated Vaccines; Biological; Biological Response Modifiers; Biology; Brain; CD8B1 gene; Chronic; Complex; Cyst; Data; Dissection; Dose; Encephalitis; Epitopes; Future; Genes; Human; Immune; Immune response; Immunity; Immunologic Surveillance; improved; Individual; Infection; Infection prevention; innovation; Interferons; Intervention; Knock-out; knockout gene; Knowledge; Life; Maps; Measures; Mediating; MHC Class I Genes; Modeling; Mus; mutant; novel strategies; Opportunistic Infections; Outcome; Parasites; pathogen; Patients; Population; prevent; Production; public health relevance; Pyrimidine; Reagent; Recrudescences; Recurrence; Research; research study; Risk; Role; Rupture; T cell response; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; tool; Toxoplasma gondii; Toxoplasmosis; Vaccinated; Vaccination; Vaccine Design; vaccine development; vaccine-induced immunity; Vaccines; Virulent; Work
Relevance: This project is relevant to understanding immune control of acute and chronic infection in the AIDS opportunistic pathogen Toxoplasma gondii. No vaccine is currently available to prevent infection, and no treatment is available to eradicate chronic infection in infected individuals. By addressing fundamental and functional aspects of immune control in T. gondii infection in this project, we expect to reveal new information that can be directly applied to vaccine development to prevent infection, as well as to identify potential interventions to eliminate chronic infection in already infected people who are particularly at risk of serious recurrent infection(s)
Project start date: 2010-07-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-09-164
5R21AI091461-02 (2011): $234630
T. GONDII: PYRIMIDINE SYNTHESIS AS A CHEMO TARGET
J David, Professor
Dartmouth Collegecity: Hanover country: United States (us)
Grant 5R01AI041930-13 from National Institute Of Allergy And Infectious Diseases
Abstract: Toxoplasma gondii chronically infects approximately 30% of the USA population and causes severe life-threatening Toxoplasmic encephalitis infections in AIDS. Current drug treatments are not well tolerated, and because these treatments have little effect on the slow growing bradyzoite stages, latent parasites remain as a source of recrudescing infection in AIDS. More effective therapies that can target both tachyzoite and bradyzoite stages are urgently needed to treat acute infections as well as to prevent the toxoplasmosis caused by recrudescing parasites. Our preliminary studies have demonstrated that disruption of carbamoyl phosphate synthetase II (CPSII) in the de novo pyrimidine synthesis pathway causes a severe uracil auxotrophy with a corresponding loss of parasite growth and virulence. Consequently the pyrimidine biosynthetic pathway appears to be an excellent target for intervention, and fundamental studies on pyrimidine acquisition have the potential to identify new targets. Due to the need for uridine monophosphate in all life stages, we hypothesize that pyrimidine biosynthesis is necessary to sustain rapid tachyzoite replication as well as the viability of bradyzoites within tissue cysts. We propose to use a combination of genetic, biochemical, and cell biological approaches to better understand the fundamental biology of de novo pyrimidine synthesis and salvage pathways in T. gondii. Inhibitors of pyrimidine biosynthesis will be identified based on a screening strategy of pyrimidine auxotrophy. The information and reagents developed in our studies will be used to validate significant drug targets in the pathway as well as to validate the target(s) of inhibitors identified in this project. These studies will contribute to the development of new therapeutic agents to treat acute infections and may contribute to the development of the first therapeutic agents capable of clearing chronic infection to eliminate reactivation Toxoplasmic encephalitis in AIDS. This research project will validate drug targets and identify inhibitors of pyrimidine biosynthesis in Toxoplasma gondii using fundamental genetic, biochemical, and drug screening approaches. Consequently, this research project will accelerate the discovery of new vaccines and treatments for devastating opportunistic parasite infections in AIDS
Keywords: Acquired Immunodeficiency Syndrome; Acute; Address; Alleles; Anabolism; Anti-HIV Agents; Attenuated; auxotrophy; base; Biochemical; Biological; Biological Assay; Biology; burden of illness; Carbamyl Phosphate; Carboxy-Lyases; Cells; Chemicals; chemotherapy; Chronic; Complementary DNA; Cyst; Development; dihydroorotate; Dihydroorotate dehydrogenase; Disease; Dissection; DNA; Drug Delivery Systems; drug development; Drug usage; effective therapy; Encephalitis; Enzyme Inhibitor Drugs; Enzyme Inhibitors; enzyme pathway; Enzymes; Future; Genetic; Goals; Growth; Growth and Development function; high throughput screening; Immune response; improved; in vivo; Infection; inhibitor/antagonist; Intervention; knockout gene; Lead; Life; Ligase; Maps; mutant; novel therapeutics; novel vaccines; orotidine; Parasites; Pathway interactions; Patients; Pharmaceutical Preparations; Plasmodium falciparum; Population; Preclinical Drug Evaluation; prevent; Prevention; public health relevance; Pyrimidine; pyrimidine metabolism; Reagent; Recombinants; reconstitution; Research Project Grants; RNA; Screening procedure; Source; Staging; Testing; Therapeutic Agents; Tissues; Toxoplasma gondii; Toxoplasmosis; Treatment Protocols; treatment strategy; Uracil; Uridine Monophosphate; Vaccines; Virulence; Work
Relevance: This research project will validate drug targets and identify inhibitors of pyrimidine biosynthesis in Toxoplasma gondii using fundamental genetic, biochemical, and drug screening approaches Consequently, this research project will accelerate the discovery of new vaccines and treatments for devastating opportunistic parasite infections in AIDS
Project start date: 1997-08-01
Project end date: 2014-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-07-070
5R01AI041930-13 (2011): $387140
COLLABORATIVE SYSTEMS FOR ANALYZING TISSUE MICROARRAYS
J David, Professor
Univ Of Med/dent Nj-r W Johnson Med Schcity: Piscataway country: United States (us)
Grant 5R01CA156386-06 from National Cancer Institute
Abstract: Tissue microarray technology holds great potential for reducing the time and cost associated with conducting investigative research in cancer biology, oncology, and drug discovery. TMA´s make it possible to construct a carefully planned array such that a 20-year survival analysis can be performed on a cohort of 600 or more patients using only a few micro-liters of antibody. However, capturing, organizing, updating, exchanging, and analyzing the data generated by this technology creates a number of significant challenges. The sheer volume of data, text, and images arising from even limited studies involving tissue microarrays can over time quickly approach those of a small clinical department. The central objective of this revised renewal application is to (1) build upon the progress made in the first phase of research by expanding the reference archive of imaged TMA specimens and correlated clinical data to include a wider scope of malignancies, tissues and biomarkers; (2) develop advanced imaging, computational and data management tools to support automated analysis of tissue microarrays in collaborative frameworks; and (3) increase dissemination of the query-enabled image archive and imaging and data management tools to the clinical and research communities for research, education and clinical decision support. The aims of the proposed project will be achieved through the development and implementation of advanced computational, imaging, and pattern recognition tools and new technologies. Tissue microarray technology holds great promise for advancing investigative research in cancer biology, oncology and drug discovery. The overarching objective of the proposed project is to develop a suite of algorithms and software tools which facilitate automated imaging, analysis, and archiving of tissue microarrays. The key computational and imaging tools that were developed in the first phase of the project including a color decomposition algorithm for analyzing the staining characteristics of the histology; image analysis tools for automatically computing the integrated staining intensity, effective staining area and effective staining intensity of expression patterns; an intelligent image archiving system; caBIG compliant data management tools; a reference library of expression signatures for more than 120, 000 imaged tissue discs originating from a mixed set of cancer tissue microarrays; and a new texture descriptor based on region covariance which was shown to provide quick, reliable performance for identifying and delineating tumor regions and performing antigen localization at the tissue level. The central objective of this revised renewal application is to build upon the progress made in the first phase of our research by (1) expanding the reference archive of imaged specimens and correlated clinical data to include a wider range of tissues, cancer types and biomarkers; (2) building upon our prior work by integrating a vendor-independent interface to the TMA analysis and data management toolset to support a full range of commercially available virtual slide formats; (3) investigating the use of a new repulsion force term to be used in conjunction with the existing internal and external energy equations for improved accuracy in delineating boundaries in regions exhibiting dense, concentrations of cells; (4) integrating a variable channel module into the segmentation algorithm and evaluate its capacity to support multi-dimensional image data; (5) building upon our successful efforts to design, develop, and evaluate a quick, reliable approach for performing unsupervised, deformable co-registration of consecutive histological sections to facilitate analysis across multiple experiments and correlate image features across adjacent sections; (6) deploying the updated software suite, data management tools and query-enabled reference archive of imaged TMA specimens to the consortium of adopter sites and assess performance using quantitative imaging experiments and a newly developed man-machine comparative analysis software toolkit. Upon completion of the project the archive of imaged specimens, computational and data management tools will be made available to the clinical and research communities as shareable resources for collaborative research, education and clinical decision support
Keywords: Algorithms; Antibodies; Antigens; Archives; Area; base; biomarker; Breast; Cancer Biology; cancer Biomedical Informatics Grid; cancer type; Cells; Characteristics; Clinical; Clinical Data; Clinical Research; cohort; Color; Communities; comparative; Computer software; computerized tools; cost; Data; Data Analyses; data management; Descriptor; design; Development; drug discovery; Educational aspects; Equation; Exhibits; Glioblastoma; Head and neck structure; Histologic; Histology; Image; Image Analysis; image archival system; Imaging Device; improved; Libraries; Lymphoma; Malignant Neoplasms; man; melanoma; Microarray Analysis; Molecular Profiling; new technology; oncology; Patients; Pattern; Pattern Recognition; Performance; performance tests; Phase; Prostate; public health relevance; Research; research study; Resources; ROC Curve; Site; Slide; Software Tools; Specimen; Staining method; Stains; statistics; Survival Analysis; System; Systems Analysis; Technology; Text; Texture; Time; Tissue Microarray; Tissues; tool; tumor; Update; Vendor; virtual; Work
Relevance: Tissue microarray technology holds great promise for advancing investigative research in cancer biology, oncology and drug discovery. The overarching objective of the proposed project is to develop a suite of algorithms and software tools which facilitate automated imaging, analysis, and archiving of tissue microarrays. The key computational and imaging tools that were developed in the first phase of the project including a color decomposition algorithm for analyzing the staining characteristics of the histology; image analysis tools for automatically computing the integrated staining intensity, effective staining area and effective staining intensity of expression patterns; an intelligent image archiving system; caBIG compliant data management tools; a reference library of expression signatures for more than 120, 000 imaged tissue discs originating from a mixed set of cancer tissue microarrays; and a new texture descriptor based on region covariance which was shown to provide quick, reliable performance for identifying and delineating tumor regions and performing antigen localization at the tissue level. The central objective of this revised renewal application is to build upon the progress made in the first phase of our research by (1) expanding the reference archive of imaged specimens and correlated clinical data to include a wider range of tissues, cancer types and biomarkers; (2) building upon our prior work by integrating a vendor-independent interface to the TMA analysis and data management toolset to support a full range of commercially available virtual slide formats; (3) investigating the use of a new repulsion force term to be used in conjunction with the existing internal and external energy equations for improved accuracy in delineating boundaries in regions exhibiting dense, concentrations of cells; (4) integrating a variable channel module into the segmentation algorithm and evaluate its capacity to support multi-dimensional image data; (5) building upon our successful efforts to design, develop, and evaluate a quick, reliable approach for performing unsupervised, deformable co-registration of consecutive histologic sections to facilitate analysis across multiple experiments and correlate image features across adjacent sections; (6) deploying the updated software suite, data management tools and query-enabled reference archive of imaged TMA specimens to the consortium of adopter sites and assess performance using quantitative imaging experiments and a newly developed man-machine comparative analysis software toolkit. Upon completion of the project the archive of imaged specimens, computational and data management tools will be made available to the clinical and research communities as shareable resources for collaborative research, education and clinical decision support
Project start date: 2004-04-01
Project end date: 2015-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01CA156386-06 (2011): $373339
SOMATOSENSORY BASIS OF SPEECH LEARNING AND CONTROL
J David, Senior Research Scientist
Haskins Laboratories, Inc.city: New Haven country: United States (us)
Grant 5R01DC004669-10 from National Institute On Deafness And Other Communication Disorders
Abstract: This work focuses on somatosensory function in speech motor learning. The long-term goal is to understand how somatosensory and auditory information are integrated in the control of speech and how somatosensory feedback on its own contributes to motor learning. The studies extend from an evaluation of the specificity of motor learning and an assessment of its somatosensory basis, to an examination of somatosensory influences on speech perception. A common procedure is adopted throughout - the use of a servo-controlled robotic manipulator to deliver precise mechanical perturbations to the jaw. There has been surprisingly little work on somatosensory function in speech and on speech learning in general. Even quite basic properties of speech motor learning such as its specificity and the extent of somatosensory involvement are largely unexplored and these form the basis of this research. These topics are central to clinical issues of training and rehabilitation as well to our basic understanding of the mechanisms of speech motor control
Keywords: Achievement; Acoustics; Adopted; Affect; Articulators; Auditory; base; Brain Stem; Classification; Clinical; Cutaneous; Equilibrium; Evaluation; Feedback; Goals; Jaw; Jordan; Learning; Link; Mechanics; Modality; Motor; motor control; motor learning; Movement; Perception; Play; Positioning Attribute; Procedures; Production; Property; Rehabilitation therapy; Relative (related person); Research; response; Robotics; Role; Sensory; Series; somatosensory; sound; Specificity; Speech; Speech Perception; Speech Sound; Training; Work
Project start date: 2000-12-01
Project end date: 2012-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5R01DC004669-10 (2011): $489875
TOWARDS GENE THERAPY FOR ANGELMAN SYNDROME
J David, Associate Professor
University Of California Daviscity: Davis country: United States (us)
Grant 5R21NS071028-02 from National Institute Of Neurological Disorders And Stroke
Abstract: This proposal will develop of a potential gene therapy for Angelman syndrome (AS). The root cause of AS is the loss of the maternal copy of the gene UBE3A. In most regions of the brain, the paternal allele is intact, but is epigenetically silenced. Our goal is to activate of the silenced UBE3A allele to ameliorate the clinical manifestations of AS. We are developing zinc finger-based artificial transcription factors (ATFs) designed to activate the expression of the murine Ube3a in cultured mouse neuronal cells. In Specific Aim 1, we will create ATFs that directly activate the transcription of murine Ube3a or repress the expression of murine Ube3a-Antisense Transcript (ATS). The milestone goal will be to create one or more ATFs that can activate Ube3a expression at least 2-fold. Several ATFs have been constructed and tested, and our preliminary data show that several are able to upregulate endogenous Ube3a protein levels. In Specific Aim 2, the ATFs will be packaged into AAV viral vectors and delivered into the brains of AS mice. We will examine the distribution and expression of the ATF in the brain, its effect on Ube3a expression. The milestone goal is to achieve widespread neuronal transduction and expression of the ATF in injected brain structures (>10% of hippocampal cells, and transduction of some surrounding tissue). One or more ATFs are expected to cause at least a 2-fold increase in Ube3a expression from the paternal allele. Successful achievement of these milestones would justify expanded pre-clinical studies and the development of ATFs to the human UBE3A. Angelman Syndrome (AS) is a neurogenetic disorder causing developmental delays and neurological impairments such as lack of speech. Currently, there is no cure and no specific drug therapy. Activation of the silenced paternal UBE3A allele by an ATF could address the fundamental genetic deficiency causing AS, and thus be a potentially curative gene therapy. This proposal will investigate such a therapy in a mouse model
Keywords: Achievement; activating transcription factor; Address; Alleles; Angelman Syndrome; base; Behavioral; Brain; Brain region; Cells; cellular transduction; Clinical; Data; design; Development; Developmental Delay Disorders; Disease; Epigenetic Process; gene therapy; Genes; Genetic; Genetic Transcription; Goals; Hippocampus (Brain); Human; Impairment; Knockout Mice; Measures; Methods; mouse model; Mus; neurogenetics; Neurologic; Neurons; Pharmacotherapy; Plant Roots; preclinical study; Proteins; public health relevance; Repression; Serotyping; Speech; Structure; Testing; Tissues; Transcript; transcription factor; vector; Viral; Viral Vector; Zinc Fingers
Relevance: NARRATIVE Angelman Syndrome (AS) is a neurogenetic disorder causing developmental delays and neurological impairments such as lack of speech. Currently, there is no cure and no specific drug therapy. Activation of the silenced paternal UBE3A allele by an ATF could address the fundamental genetic deficiency causing AS, and thus be a potentially curative gene therapy. This proposal will investigate such a therapy in a mouse model
Project start date: 2010-09-15
Project end date: 2012-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PAR-08-232
5R21NS071028-02 (2011): $212492
LENTIVIRAL GENE THERAPY FOR WISKOTT-ALDRICH SYNDROME
J David, Head, Section Of Immunology
Seattle Children´s Hospitalcity: Seattle country: United States (us)
Grant 5R01AI071163-04 from National Institute Of Allergy And Infectious Diseases
Abstract: This proposal seeks to develop an effective and safe pre-clinical model for gene therapy in Wiskott- Aldrich Syndrome (WAS). While transplantation using HLA-matched bone marrow can be curative for young WAS patients, the success rate falls precipitously with increasing age. Multiple lines of evidence document a strong selective advantage for WASP expressing hematopoeitic cell subsets suggesting that introduction of the normal WASP gene into hematopoietic stem cells (HSC) could provide a viable therapeutic alternative in disease management. While conceptually simple, development of a safe and effective strategy for WASP gene replacement requires extensive pre-clinical modeling in human and animal systems. This proposal takes advantage of combined expertise, and a network of important research and clinical collaborators, to establish a lentiviral delivery system for the definitive genetic treatment of WAS. We will test the hypotheses that 1) WASP activity is crucial for both the generation of marginal zone (MZ) B cells and homeostasis of functional T-regulatory cells (TR); and that these observations help to explain the susceptibility to infection with encapsulated bacteria, and the high-incidence of autoimmunity in WAS patients, respectively. Further, we predict that LV gene therapy will rescue these key defects. 2) Lentiviral vectors containing a pan-hematopoeitic or selected lymphoid restricted promoters will lead to functional correction of lymphoid development, activation, and survival; platelet turnover; and immune function in vivo in an animal model of WAS. 3) Analysis of viral marking and expression in a non-human primate model will allow us to define the optimal vector for use in human clinical trials; and provide key data with regard to any potential toxicity of this vector and/or dysregulated WASP expression within HSC and their progeny. Our proposed studies will provide nearly all of the key expression, efficacy, and safety data required to move forward with a human gene therapy trial for WAS; and have a very high likelihood for translation into new therapies
Keywords: Address; Adoptive Cell Transfers; Age; Animal Model; Animals; Autoimmune Process; Autoimmunity; B-Lymphocytes; Bacteria; base; Biological Assay; Blood Platelets; Bone Marrow; Cell Line; Cell Lineage; Cells; Clinical; Clinical Trials; Data; Defect; Development; Disease; Disease Management; Encapsulated; Enhancers; Exhibits; experience; falls; gene correction; gene delivery system; gene replacement; gene therapy; Generations; Genes; Genetic; Goals; Half-Life; Hematopoietic; Hematopoietic stem cells; HIV; Homeostasis; Human; Human Activities; immune function; Immunologics; In Vitro; in vitro Model; in vivo; Incidence; Individual; Infection; Kinetics; knockout animal; Knockout Mice; Lead; Lentivirus Vector; Life; Lymphoid; Mediating; Modeling; Molecular Profiling; Morbidity - disease rate; Mus; nonhuman primate; Patients; Peripheral; Platelet Count measurement; Pre-Clinical Model; Predisposition; Promotor (Genetics); Proteins; Regulatory T-Lymphocyte; Research; research study; Role; Safety; Site; Streptococcus pneumoniae; success; System; T-Lymphocyte; Testing; Therapeutic; Toxic effect; Translations; Transplantation; vector; Viral; Wiskott-Aldrich Syndrome; Work
Project start date: 2008-03-01
Project end date: 2013-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01AI071163-04 (2011): $665452
2012 CSHL PTEN PATHWAYS AND TARGETS CONFERENCE
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 1R13AG042312-01 from National Institute On Aging
Abstract: This proposal seeks support for the meeting on "PTEN Pathways and Targets" to be held at Cold Spring Harbor Laboratory March, 2012. The meeting will assemble leaders in the field, together with junior faculty, postdoctoral fellows and graduate students, to discuss new, cutting-edge developments in the regulation and function of the PTEN signaling cascade in cancer, other diseases and normal development. . PTEN is one of the most commonly lost or mutated tumor suppressor genes in human cancer. PTEN is the central negative regulator for phosphoinositide 3-kinase (PI3K) signaling, a ubiquitously expressed and evolutionarily conserved pathway that transduces signals for growth, proliferation and survival, as well as additional specialized context-dependent functions. Aberrant regulation of this pathway has profound consequences on normal development and in diseases including cancer, diabetes and autism. Topics to be discussed for the 2012 meeting include 1) Upstream regulation of PTEN - molecules and mechanisms; 2) Downstream targets of PTEN signaling 3) mTORC1 and intersecting pathways 4) Role of PTEN in development and disease 5) Clinical use of PI3K pathway inhibitors. The meeting will embrace diverse technical approaches including cell biology, genetics and biochemistry, and will showcase a range of experimental systems from mammals to invertebrate model organisms. An important mission of the conference is to span the full spectrum of this field from biochemical characterization of the regulation, specificity and function of pathway components to in vivo investigations of tumor suppression. The molecular pathogenesis of cancer will be highlighted, however, dysfunction of PI3K/PTEN signaling leading to developmental defects and other disease states will also be included to provide broader perspectives and insights. Each session will be chaired by a leading scientist in the field. Oral presentations will be selected from submitted s by the session chairs in consultation with the organizers. Selected speakers will include graduate students, postdoctoral fellows and junior faculty aiming for maximal inclusion of young investigators. Of special importance are the two poster sessions, where many participants can present their work in an atmosphere conducive to informal discussion. Additional senior investigators will be invited to highlight important or rapidly moving areas. The meeting will be of moderate size and we expect about 200 people to attend, the vast majority of whom will be presenting a poster or talk. This application requests funding to support a conference at Cold Spring Harbor Laboratory on "PTEN Pathways and Targets" Tumor suppressors are genes that are protective against the development of cancer. Extensive previous research has shown that PTEN is one of the most commonly lost or mutated tumor suppressor genes in many human cancers. The conference will focus on cutting-edge research presentations from a mixture of senior leaders in the field and junior non-established scientists, and aims to provide a general discussion forum for the emerging and converging research efforts linking disruption of PTEN pathways and targets to cancer and development
Keywords: ing; Animal Model; Area; Autistic Disorder; base; Biochemical; Biochemistry; Biological Models; Biology; Caring; Cellular biology; Chemicals; Child; Clinical; Collaborations; Complex; Consultations; Data; Defect; Development; Diabetes Mellitus; Disease; Drug Industry; Ensure; Environment; Europe; Faculty; Fostering; Functional disorder; Funding; Genetic; Goals; graduate student; Growth; Human; in vivo; inhibitor/antagonist; insight; interest; International; Invertebrates; Investigation; Japan; Laboratories; Link; Malignant Neoplasms; Mammals; meetings; Minority; Mission; Molecular; Mutate; Oral; Participant; Pathogenesis; Pathway interactions; Phosphatidylinositols; Phosphotransferases; Physiology; planetary Atmosphere; Play; Postdoctoral Fellow; posters; PTEN gene; Regulation; Request for Applications; Research; Research Institute; Research Personnel; Role; role model; Scientist; Senior Scientist; Services; Signal Pathway; Signal Transduction; Specificity; structural biology; success; symposium; System; Tumor Suppression; Tumor Suppressor Genes; Woman; Work
Relevance: This application requests funding to support a conference at Cold Spring Harbor Laboratory on "PTEN Pathways and Targets" Tumor suppressors are genes that are protective against the development of cancer. Extensive previous research has shown that PTEN is one of the most commonly lost or mutated tumor suppressor genes in many human cancers. The conference will focus on cutting-edge research presentations from a mixture of senior leaders in the field and junior non-established scientists, and aims to provide a general discussion forum for the emerging and converging research efforts linking disruption of PTEN pathways and targets to cancer and development
Project start date: 2011-09-30
Project end date: 2012-08-31
Budget start date: 30-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-10-071
1R13AG042312-01 (2011): $7500
COLD SPRING HARBOR LABORATORY CONFERENCE ON PHARMACOGENOMICS & PERSONALIZED THERA
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 5R13GM096522-02 from National Institute Of General Medical Sciences
Abstract: This proposal is a request for partial financial support for a conference on PHARMACOGENOMICS & PERSONALIZED THERAPY to be held November 17 - 21, 2010 at Cold Spring Harbor Laboratory. This represents the eighth in a series of joint Cold Spring Harbor/Wellcome Trust conferences on Pharmacogenomics held in Hinxton, UK and Cold Spring Harbor in alternate years. This conference will focus on the opportunities presented by the growing contribution of emerging genomic information and technologies to interdisciplinary approaches in the study of variable responses of humans to drugs and toxic agents, and how research may benefit the individual. The meeting will provide an in depth focus on diverse areas including mechanisms in pharmacogenomics; contemporary advances in pharmacogenomics; basic genomics - DNA sequencing and copy number variation; whole-genome associations & translational bioinformatics; cardiovascular pharmacogenomics; cancer pharmacogenomics; clinical application of pharmacogenomics; & public-private partnerships. The conference will bring together senior and junior investigators, postdoctoral and postgraduate researchers, medical, regulatory and ethical experts in a range of disciplines to share existing research and experience, and to explore the potential of working together in new international and interdisciplinary research areas for the benefit of individual patients. Co-chairs and a limited number of invited speakers will provide critical focus for the conference, while the remainder of the talks will be selected from the s on the basis of scientific merit and relevance. This balance of talks allows the conference to feature presentations by leading scientists, to be responsive to exciting new developments, to encourage diverse participation and to recognize new investigators. Cold Spring Harbor Laboratory and the meeting organizers will make particular efforts to encourage attendance and participation by women and minority scientists. The subsequent meetings (2012 and 2014) will follow a similar format and will include topics highly relevant to the current research at the time of the meeting. This proposed conference on Pharmacogenomics & Personalized Therapy is intended to have a strong multidisciplinary focus and to address diverse issues related to applying genomics to variable drug response in the human population. This conference is intended to provide a format for the exchange of ideas and information and to discuss the latest research findings and technical advances towards the study of pharmacogenetics and pharmacogenomics
Keywords: ing; Address; Area; base; Bioinformatics; Biological Models; Biotechnology; Cardiovascular system; clinical application; Copy Number Polymorphism; Data; Development; Discipline; DNA Sequence; Ensure; Environment; Equilibrium; Ethics; European; experience; Faculty; Financial Support; Fostering; Genome; Genomics; graduate student; Health; Human; Individual; Industry; Information Technology; interdisciplinary approach; Interdisciplinary Study; interest; International; Joints; Laboratories; Malignant Neoplasms; Medical; meetings; Minority; multidisciplinary; Oral; Participant; Patients; Pharmaceutical Preparations; Pharmacogenetics; Pharmacogenomics; Pharmacologic Substance; Population; Postdoctoral Fellow; postdoctoral investigator; posters; public-private partnership; Research; Research Institute; Research Personnel; response; Science; Scientist; Series; success; symposium; Time; Trust; Woman; Work
Relevance: This proposed conference on Pharmacogenomics & Personalized Therapy is intended to have a strong multidisciplinary focus and to address diverse issues related to applying genomics to variable drug response in the human population. This conference is intended to provide a format for the exchange of ideas and information and to discuss the latest research findings and technical advances towards the study of pharmacogenetics and pharmacogenomics
Project start date: 2010-09-30
Project end date: 2015-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-10-071
5R13GM096522-02 (2011): $1
MECHANISMS OF DRUG ACTION AND DISPOSITION
J David, Professor
University Of Texas Sw Med Ctr/dallascity: Dallas country: United States (us)
Grant 2T32GM007062-37 from National Institute Of General Medical Sciences
Abstract: This program will provide integrated pre-doctoral training in the pharmacological sciences for 13 students in the Division of Basic Sciences (DBS), the interdisciplinary graduate curriculum of the University of Texas Southwestern Medical Center. Graduates of the program will be well-prepared to pursue postdoctoral training or other career paths that lead to independent research programs focused on molecular and cellular approaches to understanding the mechanism of action of drugs, hormones, and other regulatory molecules. Students with strong undergraduate training in physical and biological science will receive interdisciplinary instruction in pharmacology, biochemistry, molecular and cell biology, and physiology. Advanced didactic training, student seminars, journal clubs, research rotations, and dissertation research projects will complete the research experience. Students will also receive opportunities to present their work orally or in posters at national and international meetings. All students will be encouraged to finish their training within 4-5 years of matriculating. Trainees Students who have completed the integrated first year curriculum of the DBS or within the first two years of our M.D. /Ph.D. training program will be considered for appointment as a trainee by the Steering Committee. Most students participate in the Cell Regulation graduate program of the DBS. Selection will be based on a student´s undergraduate and graduate performance and on the commitment of the student and his/her mentor to pursue a course of training consistent with goals of the program. Particular emphasis will be placed on candidates that the Steering Committee feels are "diamonds-in-the-rough" and show potential beyond their didactic training credentials. Faculty The 61 members of the training faculty come from 18 different Departments, 3 different Centers, and represent 9 interdisciplinary graduate programs of the DBS. These individuals bring a wealth of experience (many are Nobel laureates and National Academy members) and provide substantial diversity in their approaches to problems of pharmacological interest. Senior, mid-level, and junior faculty are represented, and the UT Southwestern Endowed Scholars program continues to provide a yearly influx of talented new junior faculty to the program. Over the past decade there has been a paucity of new therapeutic agents to treat human diseases. With recent technological advances in DNA sequencing and the ability to map intra- and intercellular regulatory networks in great detail, the potential to discover and characterize novel therapeutics has never been more promising. This program seeks to train the next generation of scientists who will make these discoveries and insure that scientific research on human health remains vibrant and at the cutting edge
Keywords: Drug effect disorder
Relevance: Over the past decade there has been a paucity of new therapeutic agents to treat human diseases. With recent technological advances in DNA sequencing and the ability to map intra- and intercellular regulatory networks in great detail, the potential to discover and characterize novel therapeutics has never been more promising. This program seeks to train the next generation of scientists who will make these discoveries and insure that scientific research on human health remains vibrant and at the cutting edge
Project start date: 1975-07-01
Project end date: 2016-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-10-036
2T32GM007062-37 (2011): $343622
ROLE OF PLA2R AND ANTI-PLA2R IN IDIOPATHIC MEMBRANOUS NEPHROPATHY
J David
Boston Medical Centercity: Boston country: United States (us)
Grant 1R01DK090029-01A1 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: This program will address three questions in order to better understand the role of the M-type phospholipase A2 receptor (PLA2R) as a major target antigen in idiopathic membranous nephropathy (MN) and the properties of the anti-PLA2R antibodies that are specifically detected in patients with the disease. 1. Do variants in the PLA2R1 gene account for susceptibility to idiopathic MN? This aim is based on the finding that the PLA2R epitope identified by anti-PLA2R autoantibodies is conformation dependent; the known propensity for other members of the mannose receptor/PLA2R family to exist in bent or extended configurations; differences in immunoreactivity between PLA2R in normal and MN kidney tissues; and the presence of several non-synonymous SNPs in PLA2R1, including eight within the N- terminal region that contains the epitope, at least three of which are predicted to affect PLA2R structure. Hypothesis Variants in PLA2R1 may explain the unique properties of the pathogenic epitope in MN. Approach Genomic variants in PLA2R1 from patients with idiopathic MN will be compared to matched controls to determine if there are unique SNPs that co-segregate with the disease. Particular attention will be paid to those variants that are predicted to affect PLA2R structure or function. 2. Do anti-PLA2R autoantibodies activate complement and, if so, which IgG subclass is responsible and which pathway is activated? This aim will address the apparent paradox that IgG4, the major IgG subclass in idiopathic MN and predominant anti-PLA2R subclass, is incapable of activating the classical complement pathway, yet complement components are commonly present in the glomerular immune deposits in idiopathic MN. The presence of mannan-binding lectin (MBL) and activated C4 but absent C1q in the immune deposits suggests that the lectin pathway may be involved. It is noteworthy that immunoglobulins lacking terminal galactose on Fc N-linked glycans have been shown to activate MBL. Hypothesis IgG4 anti-PLA2R autoantibodies may activate complement via the lectin pathway. Approach The ability of PLA2R IgG subclasses to activate complement will be assessed. If IgG4 activates complement, its ability to bind and activate MBL will be determined and its glycosylation state examined. 3. Is recurrent MN in transplanted human kidneys associated with circulating anti-PLA2R? Idiopathic MN frequently recurs in the transplanted kidney and is associated with a high risk of allograft loss. There is presently no way to predict which patients are likely to recur. Hypothesis The presence of circulating anti-PLA2R will predict the recurrence of MN. Approach Pretransplant and serial post-transplant sera from patients with idiopathic MN will be tested to determine if the presence of anti-PLA2R antibodies predates and presages the recurrence of MN. Idiopathic membranous nephropathy (MN) is a relatively common worldwide cause of proteinuric kidney disease in all ethnic groups with a high rate of recurrence after kidney transplantation. Following on our finding that the M-type phospholipase receptor (PLA2R) is a specific target antigen of circulating autoantibodies in a high proportion of patients with MN, we propose to examine if there is a genetic susceptibility to the disease and determine if alterations in the sugar coating of the autoantibodies might explain their ability to cause kidney damage by activating the complement system. We also plan to find out if the likelihood of recurrence after transplantation can be predicted by detecting circulating antibodies to PLA2R
Keywords: Accounting; Address; Affect; Allografting; Antibodies; Antigen Targeting; Antigens; Attention; Autoantibodies; base; Binding (Molecular Function); Biopsy; Classical Complement Pathway; Cleaved cell; Clinical; Complement; Complement 1q; Complement 4b; Complement Membrane Attack Complex; complement system; Control Groups; Deltastab; Deposition; Development; Diagnosis; Disease; Early Diagnosis; End stage renal failure; Epitopes; Ethnic group; Family; Fc Receptor; Galactose; Genes; Genetic Predisposition to Disease; Genomics; glycosylation; high risk; Human; Idiopathic Membranous Nephropathy; IgG Receptors; IgG1; IgG3; IgG4; Immune; Immunoglobulin G; Immunoglobulins; immunoreactivity; in vivo; Injury; Kidney; Kidney Diseases; Kidney Transplantation; Korea; Lectin; Link; Mannose Binding Lectin; mannose receptor; Mediating; member; Membranous Glomerulonephritis; Molecular Conformation; N-terminal; Oligosaccharides; Pathway interactions; Patients; Phospholipase; Phospholipase A2; podocyte; Polysaccharides; Positioning Attribute; Predisposition; programs; Property; Proteins; Proteinuria; receptor; Recurrence; Reducing Agents; Relative (related person); Reporting; Resources; Role; Serum; Structure; sugar; Taiwan; Testing; Time; Tissues; Transplantation; Urine; Variant
Relevance: Idiopathic membranous nephropathy (MN) is a relatively common worldwide cause of proteinuric kidney disease in all ethnic groups with a high rate of recurrence after kidney transplantation. Following on our finding that the M-type phospholipase receptor (PLA2R) is a specific target antigen of circulating autoantibodies in a high proportion of patients with MN, we propose to examine if there is a genetic susceptibility to the disease and determine if alterations in the sugar coating of the autoantibodies might explain their ability to cause kidney damage by activating the complement system. We also plan to find out if the likelihood of recurrence after transplantation can be predicted by detecting circulating antibodies to PLA2R
Project start date: 2011-09-01
Project end date: 2016-07-31
Budget start date: 1-SEP-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-10-067
1R01DK090029-01A1 (2011): $422500
Sponsored Links Excellgen http://Excellgen.com
TUBERCULOSIS INFECTION AND GENETIC VARIATION
J David, Senior Fellow
University Of Washingtoncity: Seattle country: United States (us)
Grant 5K23AI085036-02 from National Institute Of Allergy And Infectious Diseases
Abstract: This K23 award will provide an opportunity for Dr. Horne to develop as an independent investigator in patient-oriented clinical research on genetic variation in innate immunity and susceptibility to tuberculosis (TB) infection. This career development application describes a comprehensive plan to accomplish the following goals 1) assemble a cohort of patients in order to identify genetic, exposure-, and pathogen-related risk factors for TB infection, 2) develop expertise in host genetic variation and TB infection, 3) develop an understanding of the host innate immune response following an exposure to Mycobacterium tuberculosis (MTb), and 4) develop an independent clinical research career. These goals will be accomplished through mentorship by key personnel, didactic course work, instruction in practical laboratory skills, and participation in scientific meetings. We will assemble a cohort of patients in Seattle, Washington, who have a known TB exposure with the following specific aims 1) thoroughly characterize clinical and exposure-related factors, identify MTb phylogenetic lineage from the index case, and use these factors to develop a risk factor model for TB infection; and 2) perform a nested case- control study in unrelated subjects to determine whether single nucleotide polymorphisms (SNPs) in candidate genes related to innate immunity are associated with TB infection. In our analysis of genetic associations, we will adjust for significant clinical, exposure and pathogen-related risk factors identified in Aim 1. We will confirm genetic associations in a previously collected family- based cohort. This study is novel for the outcome of TB infection and the inclusion of MTb lineage. The rich academic environment at the University of Washington is ideal for Dr. Horne´s training and has allowed him to assemble an advisory committee whose members possess expertise in innate immunology, MTb phylogenetics, genetic epidemiology, biostatistics, risk factor modeling and TB transmission. Dr. Hawn, the primary mentor, is a successful and recognized expert in innate immunity, especially pathogen recognition receptors and associated pathways. He has investigated the role of Toll-like receptors (TLRs) in mycobacterial infection in functional and genetic association studies. Dr. Peter Small, co-mentor, is an internationally recognized expert in MTb phylogenetics. /Relevance TB infection, compared to TB disease, is an understudied area and little is known about host genetics and susceptibility to TB infection. TB is estimated to infect one-third of the global population and findings from this study could be important to TB vaccine development. Findings may also have relevance to other infectious diseases
Keywords: Advisory Committees; Alveolar Macrophages; Area; base; Biological Assay; Biometry; Candidate Disease Gene; career; career development; case control; Cells; Clinical; Clinical Research; cohort; Communicable Diseases; Complex; design; Disease; Disease Association; DNA; Environment; Evaluation; experience; Exposure to; Family; Future; gene environment interaction; Genes; Genetic; genetic association; genetic epidemiology; Genetic Polymorphism; Genetic Predisposition to Disease; genetic risk factor; Genotype; Goals; Host Defense; Human; Human Resources; Immune response; Immune system; Immunologic Receptors; Immunologics; Immunology; indexing; Individual; Infection; Instruction; Interferon Type II; Investigation; Laboratories; Lung; macrophage; meetings; member; Mentored Patient-Oriented Research Career Development Award; Mentors; Mentorship; Modeling; Mycobacterium Infections; Mycobacterium tuberculosis; Natural Immunity; Nested Case-Control Study; novel; Outcome; pathogen; pathogen exposure; Pathway interactions; patient oriented; Patients; Phylogenetic Analysis; Population; Predisposition; Process; prophylactic; prospective; receptor; Research; Research Infrastructure; Research Personnel; Risk Factors; Role; Single Nucleotide Polymorphism; skills; Target Populations; Toll-Like Receptor 1; Toll-like receptors; Training; transmission process; Tuberculosis; Tuberculosis Vaccines; Uganda; Universities; vaccine development; Variant; Variation (Genetics); Washington; Work
Project start date: 2010-05-01
Project end date: 2015-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-09-043
5K23AI085036-02 (2011): $127521
1K23AI085036-01 (2010): $127521
VASCULAR FUNCTION, COGNITION, AND BRAIN MRI IN ATHEROSCLEROTIC VASCULAR DISEASE
J David
University Of Iowacity: Iowa City country: United States (us)
Grant 5R01AG030417-04 from National Institute On Aging
Abstract: This is the third submission of an R01 requesting five years of funding to support research on vascular disease, cognition, and brain structure in individuals with atherosclerotic vascular disease (AVD) and healthy comparison participants. AVD is the leading cause of morbidity and mortality in industrialized nations and is a primary or contributing factor in well over half of all cases of dementia. Despite this, there are relatively few longitudinal studies of AVD-related cognitive decline and the factors that are most predictive of this process. The early identification of individuals who are at greatest risk for such decline will have important implications for the development of interventions aimed at preventing or attenuating this important health problem. Our research team is using an innovative physiological measure (forearm vascular function - the degree to which forearm vessels dilate in response to vasoactive agents) as an indicator of vascular health to be examined in relation to cognition and brain structure. We have published cross-sectional findings in Stroke and in Arteriosclerosis, Thrombosis, and Vascular Biology showing that this measure of vascular function is significantly related to cognition in patients with AVD, and we have obtained pilot data that suggest that this measure is also related to frontal/temporal grey and white matter volumes. The proposed research will significantly expand upon our current findings by allowing us to achieve the following goals 1) Determine whether the relationship between vascular function and cognition is unique to AVD or is also present in the general elderly population, as part of the normal aging process; 2) Determine whether vascular function measured at baseline, and change in vascular function across time, are associated with cognitive decline across time; 3) Determine the relationships among oxidative stress-mediated suppression of vascular function, neurohumoral mediators of vasoconstriction (endothelin levels, indicators of renin-angiotensin- aldosterone system and sympathetic nervous system activation), vascular function, and cognition; 4) Determine the relationship between vascular function and specific MRI-based brain measures (both cross- sectionally and longitudinally) in patients with AVD; 5) Conduct a pilot study on vascular function and MRI- based brain measures in healthy comparison participants. We will study healthy elderly participants (N = 100) as well as those with AVD (N = 135, 85 of whom are already enrolled in our ongoing research). This will allow us to conduct baseline and three-year follow-up assessments on those participants who are newly enrolled, and six-year follow-up assessments on those who are already participating in our research. The proposed research will allow us to make significant strides toward our long-term goals of elucidating the mechanisms by which vascular disease contributes to cognitive decline, finding better ways to identify those individuals who are at greatest risk for vascular-related cognitive decline, and developing more effective treatments to prevent or attenuate this process. Atherosclerotic vascular disease is a major contributing factor to cognitive decline and dementia in the elderly. This study is designed to help clarify the mechanisms by which vascular disease causes cognitive decline and to develop better ways to identify those individuals who are at greatest risk for vascular-related cognitive changes
Keywords: Accounting; Address; aging population; Aging-Related Process; Arteriosclerosis; Atherosclerosis; Attenuated; Base of the Brain; Biology; Blood Vessels; Brain; brain volume; Cerebrovascular Disorders; Cognition; cognitive change; comparison group; Data; Dementia; design; Developed Countries; Early identification; effective therapy; Elderly; Endothelin; Enrollment; follow-up; Forearm; Functional disorder; Funding; Goals; Grant; gray matter; Health; Hippocampus (Brain); Impaired cognition; Individual; innovation; Longitudinal Studies; Magnetic Resonance Imaging; Measures; Mediating; Mediator of activation protein; Morbidity - disease rate; Mortality Vital Statistics; neuroimaging; neuropsychological; normal aging; Oxidative Stress; Participant; Patients; Performance; Physiological; Pilot Projects; Population; prevent; Process; Publishing; Renin-Angiotensin-Aldosterone System; Research; Research Support; Resistance; response; Risk; Sampling; stroke; Structure; Sympathetic Nervous System; Temporal Lobe; therapy development; Thrombosis; Time; Vascular constriction (function); Vascular Diseases; vasoactive agent; white matter
Project start date: 2008-09-01
Project end date: 2013-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01AG030417-04 (2011): $463073
2011 EUKARYOTIC DNA REPLICATION & GENOME MAINTENANCE
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 1R13CA159756-01 from National Cancer Institute
Abstract: This conference will be the 13th biennial meeting on Eukaryotic DNA Replication and Genome Maintenance and follows the highly successful meetings that have been held at Cold Spring Harbor every second year since September 1987. It is the only regularly occurring meeting that is exclusively focused on eukaryotic DNA replication. Because of this focus, the meeting has played a major role in the rapid growth in our understanding of the eukaryotic DNA replication process and how it is integrated into the cell division cycle. This year´s conference will be devoted to fundamental research topics related to chromosome duplication, structure and function, and will include important areas of biological research in the areas of cell cycle and growth control, genomic amplification, and cancer research. Starting with the 2007 meeting, we placed an increased emphasis on the central role of DNA replication in the DNA damage and replication stress response and the strategies used by cells to minimize the threats to genomic integrity arising from DNA replication. The rapid convergence of the DNA replication and DNA damage response fields makes this a timely meeting. The format of the meeting will ensure that recent results will be communicated and discussed face-to-face, which will enhance progress and collaboration. The participation of young investigators and minority and women scientists is strongly encouraged. The 2011 meeting will include a diverse array of topics, systems, and approaches including studies of (1) chromosomal replication and gene amplification in organisms as diverse as yeast, Drosophila, Xenopus and mammalian cells; (2) the replication of viral chromosomes, including SV40, polyomavirus, cytomegalovirus, herpesvirus, papillomaviruses, and Epstein-Barr virus; (3) control of DNA replication in the cell cycle; (4) structure and function of the chromosomal elements controlling replication including origins of replication and telomeres; (5) connections between DNA replication and the cell cycle, development, and cancer; (6) mechanisms that control genomic integrity including DNA replication checkpoints and post- replication DNA repair; (7) novel approaches including genomics, systems biology, and single molecule studies. Since DNA replication is crucial to cell division, and uncontrolled growth is a hallmark of tumors, the relevance of this meeting to cancer research, and ultimately to improved therapies, cannot be overemphasized. Replication of DNA is a fundamental process in all (eukaryotic) life. Understanding the mechanism by which this process occurs, and how this process coincides with genome. The intellectual merits of this conference include the opportunity for leading investigators at all stages of their scientific careers to share and discuss their latest results and concepts. The informal peer review in oral and poster sessions is invaluable in providing rapid feedback that will fruitfully steer and accelerate future research. This conference also provides ample opportunity for learning and building collaborations - no parallel sessions are planned and so all attendees share a common experience, while the secluded venue maximizes the likelihood of productive scientific exchange. Large conferences generally can have significant impact in their field. Unique aspects of this conference that have particularly broad impact are the active participation of younger scientists who will particularly benefit from the opportunity to present their latest ideas. The conference archive allows participants to share aspects of the conference with their colleagues who were unable to attend while protecting the right of the presenting authors to present unpublished research. The duplication of the genetic material (DNA) is a process central to all unicellular and multicellular organisms. In higher plants and animals, the genetic material is packaged together with specialized proteins in structures called chromosomes that are themselves packed within the cell nucleus. How these chromosomes are duplicated, and how the DNA they contain is replicated, to allow faithful copies to be made from generation to generation, is a central question in biology. The entirety of the DNA of an organism (its "genome") has to be copied with minimal errors every time the cell divides, which involves multiple "proof-reading" and error- correction" mechanisms that operate at the molecular level and ensure excellent copying fidelity. When cells begin to divide uncontrollably during the development of cancer, errors may accumulate which may themselves contribute to how the disease progresses. The centrality and importance of this process, and its relevance to the development of cancer in particular, has led a large but diverse group of scientists towards the study of DNA replication in numerous systems. The aim of the conference on Eukaryotic DNA Replication & Genome Maintenance is to provide a forum to bring together the diverse group of scientists working in different areas of this field. The conference will be an open international meeting devoted to the most recent advances in the rapidly evolving field of eukaryotic DNA replication and the maintenance of genome integrity. Oral presentations will consist of a combination of invited presentations and selected presentations from submitted s. This ensures the participation of junior and senior leaders in the field and the presentation of the most exciting results emerging at the time of the meeting. The oral presentations will be complemented by poster presentations in three poster sessions, also selected from submitted s
Keywords: ing; American; Animal Genetics; anticancer research; Archives; Area; Asians; base; Biochemistry; biological adaptation to stress; biological research; Biology; cancer cell; career; Cell Cycle; Cell Cycle Progression; Cell division; Cell Nucleus; Cells; Chromosomes; Collaborations; Complement; Cytomegalovirus; Data; Development; Disease; DNA; DNA biosynthesis; DNA Damage; DNA Repair; DNA Replication Damage; Drosophila genus; Drug resistance; Elements; Ensure; Environment; Eukaryota; European; Event; experience; Faculty; Feedback; Fostering; fundamental research; Gene Amplification; Generations; Genetic; Genetic Materials; Genome; Genomics; Goals; graduate student; Growth; Herpesviridae; Human Herpesvirus 4; improved; interest; International; Investigation; Laboratories; Learning; lectures; Length of Stay; Life; Maintenance; Malignant Neoplasms; Mammalian Cell; meetings; Minority; Mitotic Cell Cycle; Molecular; Molecular Biology; novel strategies; Oral; Organism; Papillomavirus; Participant; Peer Review; Play; Polyomavirus; Postdoctoral Fellow; posters; Process; Proteins; Published Comment; rapid growth; Reading; Regulation; repaired; Replication Origin; Replication-Associated Process; Research; Research Institute; Research Personnel; response; Role; S Phase; Schedule; Science; Scientist; Seasons; Selection Criteria; Senior Scientist; Series; Signal Transduction; Simian virus 40; single molecule; Staging; Structure; success; symposium; System; Systems Biology; telomere; Time; tumor; unpublished works; Vascular Plant; Viral; Woman; Work; Xenopus; Yeasts
Relevance: The duplication of the genetic material (DNA) is a process central to all unicellular and multicellular organisms. In higher plants and animals, the genetic material is packaged together with specialized proteins in structures called chromosomes that are themselves packed within the cell nucleus. How these chromosomes are duplicated, and how the DNA they contain is replicated, to allow faithful copies to be made from generation to generation, is a central question in biology. The entirety of the DNA of an organism (its "genome") has to be copied with minimal errors every time the cell divides, which involves multiple "proof-reading" and error- correction" mechanisms that operate at the molecular level and ensure excellent copying fidelity. When cells begin to divide uncontrollably during the development of cancer, errors may accumulate which may themselves contribute to how the disease progresses. The centrality and importance of this process, and its relevance to the development of cancer in particular, has led a large but diverse group of scientists towards the study of DNA replication in numerous systems. The aim of the conference on Eukaryotic DNA Replication & Genome Maintenance is to provide a forum to bring together the diverse group of scientists working in different areas of this field. The conference will be an open international meeting devoted to the most recent advances in the rapidly evolving field of eukaryotic DNA replication and the maintenance of genome integrity. Oral presentations will consist of a combination of invited presentations and selected presentations from submitted s. This ensures the participation of junior and senior leaders in the field and the presentation of the most exciting results emerging at the time of the meeting. The oral presentations will be complemented by poster presentations in three poster sessions, also selected from submitted s
Project start date: 2011-09-01
Project end date: 2012-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-10-071
1R13CA159756-01 (2011): $6500
J David
University Of Utahcity: Salt Lake City country: United States (us)
Grant 5T32GM007464-35 from National Institute Of General Medical Sciences
Abstract: This application requests continuing support for an interdepartmental PhD training program in the area of genetics. Training is provided by a collection of 56 faculty mentors distributed between seven departments at the School of Medicine and the College of Science. The participating faculty members are a highly interactive group which has demonstrated its ability to train students and work together productively. They work together through this training program, through the campus-wide Combined Program in Molecular Biology and through a complex network of shared teaching, journal clubs and scientific collaborations. The requested training grant continuation helps support graduate student training in the area of genetics and contributes to our ability to provide students with a broad education in all areas of genetics. The training program provides cohesion for students and faculty in the areas of genetics. The program identifies outstanding students working on some aspect of genetics, and brings them into a group with other genetics students and the collective genetics faculty where a broad range of topics in current genetics research is discussed. Their research success is promoted by multiple opportunities to talk about their projects, as well as getting critical feedback about their research. A major feature of this training program is its yearly retreat at which the entire group of trainees is brought together with faculty members and an outside speaker. At this meeting, students present their ongoing work and benefit from in-depth discussion, suggestions and critique. A series of regular presentations by both inside and outside speakers comprises a second important training activity. A successful part of this series is seminars by former students supported by this training grant, who provide valuable mentoring. Trainee selection is based primarily on demonstrated excellence. In addition, effort is made to represent the entire breadth of genetics research underway here. One of the outstanding traditions of Utah´s genetics program has been its breadth, and we promote interaction between those using mechanistic and population-based research genetic approaches. Trainees are selected from the pool of graduate students recruited and given formal course work by a Campus-wide Combined Molecular Biology Program. This program advertises our general graduate program, screens applicants and teaches a set of four courses required by all students in the program. Students admitted through this program spend their first year doing course work and four lab rotations after which they choose a thesis advisor and become administratively associated with that advisor´s department. Only after starting their thesis research and passing their qualifying examination are they eligible to apply for training grant support
Keywords: Genetic; Training Programs
Project start date: 1977-07-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5T32GM007464-35 (2011): $278746
ENGINEERING SKELETAL MUSCLE WITH BIODEGRADABLE HYDROGELS
J David, Professor Of Bioengineering
Harvard Universitycity: Cambridge country: United States (us)
Grant 5R01DE013349-11 from National Institute Of Dental & Craniofacial Research
Abstract: There exists a significant need for skeletal muscle reconstruction or regeneration in a variety of diseases or following trauma, and a new approach to skeletal muscle regeneration is proposed in this application. This approach combines the localized presentation of specific growth factors with cell transplantation using biodegradable polymer vehicles. The vehicles will serve to control the availability of the factors to host and transplanted cells, control the activation and migration of the transplanted cells to repopulate and regenerate damaged host tissue, and regulate muscle vascularization and reinnervation to enhance transplanted and host cell survival. The specific hypothesis guiding this proposal is that the regeneration of damaged muscle tissue can be controlled by providing a sustained delivery of activated satellite cells to the damaged tissue, and regionally regulating VEGF signaling to enhance their survival and participation in regeneration. This hypothesis will be addressed with the following aims (1) Determine whether sustained delivery of satellite cells leads to repopulation and regeneration of lacerated sternomastoid muscle when delivered using a vehicle designed to activate the cells (e.g., proliferate, migrate) but prevent their terminal differentiation until released. (2) Examine the ability of regionally available vascular endothelial growth factor (VEGF) to promote regeneration of lacerated sternomastoid muscle, and (3) Analyze the regeneration of lacerated sternomastoid muscle following sustained satellite cell delivery using vehicles that provide regional VEGF availability and locally regulate cell activation and migration. The successful completion of these aims will ead to new strategies for the regeneration of functional muscle tissue. This work will likely have significant applications in craniofacial reconstructive surgery, and will apply to the regeneration of muscle throughout the body. In a broader sense, this new approach to tissue regeneration, which focuses on repopulation of damaged tissues by the sustained delivery of cells competent to participate in regeneration in concert with revascularization will likely be broadly applicable to other tissues and organs in the body
Keywords: Address; Adhesives; biodegradable polymer; Cell Survival; Cell Transplantation; Cell Transplants; Cells; craniofacial; Cues; design; Disease; Engineering; Excision; Goals; Growth Factor; Hydrogels; improved; Malignant Neoplasms; migration; Minor; Modeling; Muscle; muscle regeneration; Mutation; Natural regeneration; nerve supply; novel strategies; Organ; Perfusion; prevent; Proliferating; reconstruction; Reconstructive Surgical Procedures; reinnervation; Research; Role; satellite cell; Signal Transduction; Skeletal muscle structure; stem cell niche; Structure of sternocleidomastoid muscle; tissue regeneration; Tissues; Transplantation; Trauma; Vascular Endothelial Growth Factors; Vascularization; Work
Project start date: 2000-03-01
Project end date: 2012-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
5R01DE013349-11 (2011): $268498
RADIATION PROTECTORS AND RADIATION THERAPY COUPLED CHEMOPREVENTION
J David, Professor
University Of Chicagocity: Chicago country: United States (us)
Grant 5R01CA132998-03 from National Cancer Institute
Abstract: The use of radiation and chemotherapy has resulted in steadily improving cure rates in both childhood and adult cancer patients. This therapeutic success, however, has been limited by the frequent development of secondary cancers in long term survivors. Examples of therapy-related cancers include myelodysplastic syndrome and acute myeloid leukemia. These neoplastic disorders are a consequence of the therapy induced- DNA damage in hematopoietic stem cells. Patients with potentially curable cancers represent a critical population for the development of strategies for chemoprevention of secondary malignancies. The goals of this proposal are the identification, characterization, and validation of agents that can prevent mutational damage in hematopoietic stem cells while preserving the anti-tumor efficacy of radiation and chemotherapy regimens. This novel paradigm of radioprotector-mediated chemoprevention is known as therapy coupled chemoprevention (TCC). To facilitate these studies, we will use the MLL-ELL knock-in mouse model that closely recapitulates the multistep process of transformation observed in human-related acute myeloid leukemia. These MLL-ELL knock-in mice do not develop leukemia spontaneously, but exhibit a high susceptibility to leukemia following exposure to a DNA damaging agent. This model will provide a novel and unique resource to test and validate TCC strategies for patients at risk for secondary cancers. TCC agents to be examined are amifostine and phosphonol, which each possess anti-mutagenic properties at doses 4- to 16- fold lower than those required to demonstrate classical cytoprotection. We will determine the maximum non- cytoprotective doses of these TCC agents in mice having 4 day old FSa "artificial" micro lung metastases treated with ionizing radiation and an alkylating-chemotherapeutic agent such as cyclophosphamide. Using the Hprt mutation assay, we will assess the anti-mutagenic effectiveness of TCC agents in MLL-ELL knock-in mice and their wild type counterparts following exposure to ionizing radiation and alkylating agent therapy. Using the MLL-ELL knock-in mouse model, we will also evaluate the efficacy of TCC to prevent radiation- and alkylating agent-induced mutations at the Hprt locus and the development of therapy-related acute myeloid leukemia in the same anima system. This grant addresses the growing health problem of cancer patients cured of their first cancer by radiation and chemotherapy only to be diagnosed in later years with a cancer induced by those therapies. By appropriately administering to patients at the time of their radiation and chemotherapies cancer prevention drugs that can prevent mutations and cancer development without affecting the direct killing of cancer cells, patients will continued to be cured but without the risk of developing new cancers due to their treatments
Keywords: Acids; Address; Adult; Adverse effects; Affect; Alkylating Agents; Alkylating Antineoplastic Agents; Amifostine; analog; Animal Testing; Animals; Anticarcinogenic Agents; artificial lung; base; Base Pairing; Biological Assay; Biological Availability; cancer cell; Cancer Patient; cancer prevention; cancer therapy; carcinogenesis; Carmustine; cell killing; Cells; Chemoprevention; Chemopreventive Agent; Chemoprotective Agent; chemotherapeutic agent; chemotherapy; Chemotherapy-Oncologic Procedure; Childhood; Chronic; Cisplatin; Clinical; Combined Modality Therapy; Complex; Coupled; Couples Therapy; Cyclophosphamide; Cytoprotection; Cytoprotective Agent; cytotoxic; Descriptor; design; Development; Diagnosis; Disease; DNA Damage; dosage; Dose; Drug usage; Dysmyelopoietic Syndromes; effective therapy; Effectiveness; Ethylnitrosourea; Evaluation; Exhibits; Experimental Neoplasms; Exposure to; FDA approved; Frameshift Mutation; Gene Mutation; General Population; Genomics; Goals; Grant; Guanine; Head and Neck Cancer; Health; Hematopoietic; Hematopoietic stem cells; Hour; Human; Hypoxanthines; improved; Incidence; Individual; Induced Mutation; Investigation; Ionizing radiation; irradiation; Kidney; Killings; Knock-in Mouse; Laboratories; Lateral; Lesion; leukemia; Leukemia, Myelocytic, Acute; leukemogenesis; Long-Term Survivors; Lung; Lung nodule; Lymphocyte; Malignant - descriptor; Malignant Childhood Neoplasm; Malignant neoplasm of ovary; Malignant Neoplasms; Marketing; Mediating; Metastasis Induction; Modality; Modeling; Molecular; Monitor; mouse model; Mus; Mutagenesis; Mutation; Myeloid Leukemia; Names; Neoplasm Metastasis; neoplastic; neoplastic cell; Nitrosourea Compounds; Nodule; Non-Small-Cell Lung Carcinoma; Normal Cell; novel; oncology; Oral; outcome forecast; Package Insert; Parotid Gland; Patients; Peripheral Blood Lymphocyte; Pharmaceutical Preparations; Phase; phosphorothioate; Pilot Projects; Population; Postoperative Period; Predictive Value; Predisposition; prevent; Primary Neoplasm; Process; Promotor (Genetics); Property; Protocols documentation; public health relevance; Radiation; radiation effect; Radiation therapy; Radiation-Protective Agents; Radio; Regimen; Relative (related person); research study; Resources; response; Risk; Risk Factors; Schedule; Second Primary Cancers; stem; success; System; Tail; Testing; Therapeutic; Therapeutic Agents; Therapy-Related Acute Myeloid Leukemia; Therapy-Related Acute Myeloid Leukemia and Myelodysplastic Syndrome; Time; Toxic effect; Transferase; Treatment Efficacy; Treatment Protocols; treatment strategy; Treatment-Related Cancer; tumor; tumor growth; Validation; Veins; Wild Type Mouse; Work; Xerostomia
Project start date: 2009-06-01
Project end date: 2014-04-30
PFA/PA: PA-07-070
5R01CA132998-03 (2011): $291330
INTERSENSORY PERCEPTUAL NARROWING IN HUMAN INFANTS
J David
Florida Atlantic Universitycity: Boca Raton country: United States (us)
Grant 1R01HD057116-01A2 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: The two primary channels of communication in primate species are the face and the voice. Normally, facial gestures and accompanying vocalizations provide highly redundant information because they are always spatially co-located, temporally synchronous, and specified by overlapping patterns of dynamic audible and visible information. Adults profit from the multisensory redundancy that vocalizing faces offer because they use the redundancy to integrate the separate auditory and visual streams of information into coherent perceptual experiences and because the redundancy makes it easier to detect, learn, and remember vocalizing faces. Although extant evidence indicates that the ability to take advantage of multisensory redundancy emerges early in infancy, the processes underlying its emergence are still poorly understood. Consequently, the current project investigates the development of face-voice integration in human infants and explores the novel idea that perceptual narrowing plays a critical role in the development of intersensory integration. This novel idea is based on a recent discovery by the PI & his colleagues that younger infants can integrate a broader range of faces and voices than do older infants. We have found that young infants can integrate nonnative faces and vocalizations (either monkey faces and their vocalizations or foreign visible and audible speech sounds) but that older infants do not. Our findings are striking because they are counterintuitive and contrary to predictions arising from extant theories of intersensory development. They are, however, consistent with a growing literature on narrowing effects in the speech, face, and music processing domains and suggest that narrowing is a pan-sensory, domain-general process. Consequently, the current project will investigate intersensory perceptual narrowing (IPN). It will be guided by 5 specific aims whose purpose will be to shed light on (1) the role of temporal intersensory synchrony, configural processing, gestural features, and language-general processes in IPN, (2) the general nature of IPN by testing for it across different types of integration tasks, (3) the role of experience in IPN, (4) the separate contribution of unisensory perceptual processes to IPN, and (5) whether the sensitive period for IPN is affected by multisensory redundancy. We will use the intersensory matching technique to examine face-voice integration, the habituation/test method to investigate learning and discrimination of faces and voices, and measure infant visual attention to index responsiveness. By shedding light on the perceptual and experience-dependent mechanisms that underlie the development of face-voice integration in infancy, this project will provide insights into the emergence of a critical communicative skill that is essential to adaptive functioning at the perceptual, cognitive, and social levels. This, in turn, will provide insights into the etiology of various developmental disorders (e.g., autism) and will facilitate the development of better diagnostic tools for such disorders and help develop better intervention methods to ameliorate these disorders. Autism, whose hallmark is the inability of children with this developmental disability to respond to the faces and voices of people as sources of social communication, has been growing at an alarming rate of 10-17 percent per year. Because autism and related communication and learning disorders are developmental in nature, the earlier they are diagnosed the more effectively they can be ameliorated and/or prevented. The current project will investigate the development of some critical perceptual skills that enable infants to respond to people
Keywords: Adult; Affect; Age; Age-Months; Animals; Auditory; Autistic Disorder; base; Behavioral; Birth; Child; Cognitive; Collection; Communication; Communication impairment; Conceptions; Cues; design; Detection; Development; Developmental Disabilities; developmental disease/disorder; Diagnostic; Discrimination (Psychology); Discrimination Learning; Disease; Early Diagnosis; Employee Strikes; Etiology; Event; Exhibits; experience; Face; Gender; Gestures; Human; improved; indexing; infancy; Infant; infant animal; insight; Intervention; Language; Learning; Learning Disorders; Life; Light; Literature; Measures; Mediating; Memory; Methods; Monkeys; multisensory; Music; Nature; novel; Pattern; Perception; Plastics; Play; prevent; Prevention; Primates; Process; public health relevance; Race; relating to nervous system; Research; response; Role; Sensory; Series; skills; social; social communication; sound; Source; spatiotemporal; Specific qualifier value; Speech; Speech Perception; Speech Sound; Stimulus; Stream; Tactile; Techniques; Testing; theories; Thinking, function; Time; tool; Visual; Visual attention; vocalization; Voice
Relevance: Autism, whose hallmark is the inability of children with this developmental disability to respond to the faces and voices of people as sources of social communication, has been growing at an alarming rate of 10-17 percent per year. Because autism and related communication and learning disorders are developmental in nature, the earlier they are diagnosed the more effectively they can be ameliorated and/or prevented. The current project will investigate the development of some critical perceptual skills that enable infants to respond to people¿s faces and voices as sources of socially relevant communicative information. As a result, not only will this project provide insights into the possible reasons for impaired functioning in these disorders but also a scientific basis for the design of diagnostic tools that will permit the detection of these disorders much earlier than is currently possible and ultimately their prevention
Project start date: 2011-03-01
Project end date: 2016-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-10-067
1R01HD057116-01A2 (2011): $300242
TRI-INSTITUTIONAL TRAINING PROGRAM IN COMPUTATIONAL BIOLOGY & MEDICINE
J David, Associate Professor
Weill Medical College Of Cornell Univcity: New York country: United States (us)
Grant 5T32GM083937-03 from National Institute Of General Medical Sciences
Abstract: The Tri-lnstitutional Training Program in Computational Biology and Medicine (CBM) takes advantage of the outstanding educational and research resources of Cornell University in Ithaca, NY, the Weill Cornell Medical College in New York City, and the Sloan-Kettering Institute (the research arm of the Memorial Sloan- Kettering Cancer Center) to train computational biologists in the interdisciplinary approaches they need to solve the complex problems that characterize biology and medicine. The CBM training environment is designed to address the unique challenges of training scientists in this area and is characterized by (i) coursework in both quantitative and biological sciences; (ii) research rotations to enable a well-informed thesis topic selection; (iii) journal clubs and research-in-progress series to enhance program cohesion and ensure fluency in relevant disciplines; (iv) thesis research in one of a diverse array of basic to translational computational and experimental laboratories; (v) both quantitative and biological mentorship in order to ensure training balance and breadth; (vi) encouragement to engage in hybrid computational/experimental projects to foster connections between theory and experiment. The enthusiastic support for the CBM program by the Tri-lnstitutional consortium has enabled the program to enroll and support 6 students annually. However, now that the program is well established and entering its sixth year, it is clear that it would benefit from an increase in its most vital component - outstanding students. To that end, the goal of this T32 proposal is to receive funding to increase our annual admission to 9 students. The justifications for such growth include (i) an unmet demand by CBM faculty for students; (ii) the CBM applicant pool is deep enough to allow for an increase without either lowering quality or decreasing the percentage of admitted domestic students; and (iii) a national need to train more computational biologists. By enabling such growth, T32 funding would aid the CBM program in achieving its mission of training the next generation of scientist to use computational and analytical methods, often integrated with experimental and clinical studies, to solve complex interdisciplinary problems in biology
Keywords: Computational Biology; Medicine; Training Programs
Project start date: 2009-07-01
Project end date: 2014-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5T32GM083937-03 (2011): $217128
ELECTRICAL SILENCING OF THE PULMONARY VEINS
J David, Instructor In Medicine
Massachusetts General Hospitalcity: Boston country: United States (us)
Grant 5R21HL096009-02 from National Heart, Lung, And Blood Institute
Abstract: The recent discovery of genes responsible for the potassium leak currents, the KCNK family of potassium channels, has created the opportunity to directly target arrhythmogenic triggers of atrial fibrillation, rather than merely trying to contain them through pulmonary vein isolation. Expression of the KCNK0 gene in neurons can render these cells electrically silent by effectively shorting out any depolarizing currents. We have extended this work to demonstrate that expression of KCNK0 in cardiomyocytes renders them inexcitable. We thus propose to evaluate the KCNK0 gene for its ability to silence cardiomyocytes as a treatment for atrial fibrillation in the following specific aims Specific Aim1 In vitro evaluation of KCNK0 for the electrical silencing of cardiomyocytes. This will involve assessment of biological effects in cardiomyocytes - establishing transfection efficiency targets, assessing cell viability, studying the electrophysiologic effects of heterogeneity from mosaic KCNK0 expression, and assessing possible cytotoxicity. Specific Aim 2 In vivo adenoviral delivery of KCNK0 to the anterior right atrium to create a line of block. This will involve linear delivery of the Ad.KCNK0 followed 5 days later by multielectrode recordings of electrical activity. Comparison of two gene delivery methods and dose response will be evaluated with assessment of gene transfer efficiency, toxicity, and protein levels. Specific Aim 3 In vivo adenoviral delivery of KCNK0 to silence pulmonary vein myocytes in a large animal model. This will involve baseline electroanatomic mapping, followed by KCNK0 gene delivery to the pulmonary veins, and then remapping of the pulmonary veins to document electrical silencing. Animals will also undergo atrial programmed stimulation and Holter monitoring to assess for proarrhythmia. Comparison will be made to Ad.GFP treated animals as well as animals treated with traditional radiofrequency pulmonary vein isolation. Our primary objective is to demonstrate the feasibility of electrically silencing the pulmonary veins using gene transfer as a potential treatment for atrial fibrillation in an experimental animal model. In this project, sheep be will be used to determine whether expression of a modified potassium leak current, KCNKO, by adenoviral gene therapy approach leads to loss of electrical excitability (electrical silencing) of the cardiomyocytes investing the pulmonary vein ostia
Keywords: A-V Node; Ablation; Academic Medical Centers; Address; Animal Model; Animal Models and Related Studies; Animals; Anterior; Anticoagulation; Antigenic Determinants; Apoplexy; Arrhythmia; Atrial; Atrial Fibrillation; Atrio-Ventricular Node; Atrioventricular Node; atrioventricular node; atrium; Atrium, Right; Auricle of Heart; Auricular Fibrillation; AV Node; Binding Determinants; Biological; Body Tissues; brain attack; Brain TIA; Cardiac Arrhythmia; Cardiac Atrium; Cardiac Myocytes; Cardiac Tamponade; Cardiocyte; cardiomyocyte; Cell Survival; Cell Viability; Cells; Cerebral Stroke; cerebral vascular accident; Cerebrovascular accident; Cerebrovascular Apoplexy; Cerebrovascular Stroke; Complication; Constriction, Pathologic; Constriction, Pathological; Coupled; cytotoxicity; Data; Dose; Epitopes; Esophageal; Evaluation; Experimental Animal Model; Family; Family member; Gene Delivery; gene discovery; gene product; gene therapy; Gene Transfer; Gene Transfer Clinical; Gene Transfer Procedure; Gene-Tx; Genes; Genetic Intervention; genetic therapy; Goals; Heart Arrhythmias; Heart Atrium; Heart myocyte; heart tamponade; Heterogeneity; Holter Electrocardiography; Holtmon; Hour; implantation; In Vitro; in vivo; Intervention, Genetic; Ion Channels, Potassium; K channel; K element; Lytotoxicity; Maps; Membrane Potentials; Methods; model organism; Molecular; Molecular Biology, Gene Therapy; Monitoring, Holter; Muscle Cells; Muscle Cells, Cardiac; Muscle Cells, Heart; Muscle Cells, Mature; Myocytes; Myocytes, Cardiac; Nature; Nerve Cells; Nerve Paralysis; Nerve Unit; Neural Cell; Neurocyte; neuronal; Neurons; new approaches; novel approaches; novel strategies; novel strategy; overexpression; Ovis; Pace Stimulators; Pacemakers; Phrenic Nerve; Play; Potassium; Potassium Channel; Prevalence; Procedures; programs; Programs (PT); Programs [Publication Type]; Proteins; public health relevance; Pulmonary veins; Radio Frequency Ablation; radiofrequency; Radiofrequency Ablation; Radiofrequency Interstitial Ablation; response; Rest; Resting Potentials; RF ablation; Right atrial structure; Risk; Role; Rose`s tamponade; Sheep; social role; Stenosis; Stimulators, Electrical, Pace; Stroke; stroke; Structure; Structure of atrioventricular node; Structure of phrenic nerve; success; Technology; Therapy, DNA; Time; Tissues; Toxic effect; Toxicities; Transfection; transfer of a gene; Transient Ischemic Attack; Transmembrane Potentials; University Medical Centers; Vascular Accident, Brain; Work
Relevance: Our primary objective is to demonstrate the feasibility of electrically silencing the pulmonary veins using gene transfer as a potential treatment for atrial fibrillation in an experimental animal model. In this project, sheep be will be used to determine whether expression of a modified potassium leak current, KCNKO, by adenoviral gene therapy approach leads to loss of electrical excitability (electrical silencing) of the cardiomyocytes investing the pulmonary vein ostia
Project start date: 2010-02-03
Project end date: 2012-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PA-06-181
5R21HL096009-02 (2011): $220182
Sponsored Links Excellgen http://Excellgen.com
2011 EUKARYOTIC MRNA PROCESSING CONFERENCE
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 1R13CA159737-01 from National Cancer Institute
Abstract: The proposed conference on "Eukaryotic mRNA Processing" will convene scientists studying various aspects of mRNA processing, transport, RNA interference, informatics and turn-over. Major advances have recently been made in these areas, and the proposed conference will be a timely event for discussing the latest unpublished results and exchanging ideas, thereby fostering new developments in this rapidly moving field. The proposed 2011 conference will be held in August and is the eighth meeting of a conference that is held every other year at Cold Spring Harbor Laboratory. The most recent meeting was held in August 2009 and attracted 335 scientists internationally, who are actively investigating various aspects of messenger RNA maturation in eukaryotic cells, using genetic, biochemical, molecular, and cell biological approaches. As in the previous meetings, a major focus will be on nuclear events in mRNA maturation, particularly mRNA splicing and polyadenylation, but the scope will now be expanded to encompass both the impact of the history of an mRNA on its ultimate biological fate, the emerging fields of RNA interference and microRNA function as well as the application of informatics and genome-wide approaches to the analysis of RNA processing. The meeting format will consist of eight plenary sessions and two poster sessions. For the 2011 meeting we will subdivide each plenary session into two parts (separated by a coffee break) that focus on related but separate topics. This subdivision allows us to specifically seek out a broader representation of fields and to recruit a more diverse set of faculty as session chairs. As always, all speakers will be selected on the basis of the submitted s, which will encourage active participation by junior scientists. We will particularly encourage presentation of unpublished work by the students and postdoctoral fellows who are leading these projects, as has traditionally been a hallmark and a unique strength of the Cold Spring Harbor meetings. In genes of higher organisms from yeast to man, the information encoded in the DNA sequence is interrupted by non-coding regions called introns. An RNA copy of the gene has to be read off, cut and then spliced back together to remove the introns and produce a continuous "message" with the correct information to produce a protein. In many cases, the message can be cut and put back together in different combinations giving rise to proteins with different functions. This means that the number of different proteins in a cell can be much greater than the number of different genes. Mistakes in the splicing of the RNA cause serious problems as defective proteins are produced, and this sometimes happens as a consequence of genetic defects or disease. The molecular machinery that carries out this gene splicing is very complicated and has to be tightly regulated in the cell. Many scientists are studying how this messenger RNA processing occurs and is controlled, helped by the fact that the RNA splicing machinery is highly conserved between yeast and humans. This conference brings these scientists together to discuss their latest results. A key to the success of the conference is that the majority of oral presentations are given by graduate students, postdoctoral fellows and junior faculty chosen on the basis of scientific merit, ensuring that the conference showcases the latest developments which are often yet to be published. Participants come from academic centers, research institutes and industrial centers around the world to present and discuss their findings. Importantly, this application requests support for junior scientists who might not otherwise be able to attend to actively participate in the meeting
Keywords: ing; American; Animal Model; Architecture; Area; Asians; Back; base; Binding (Molecular Function); Biochemical; Biochemical Genetics; Biochemistry; Biological; Cells; Coffee; Data; Development; Disease; disease-causing mutation; Dissection; DNA Sequence; Drosophila genus; Emerging Technologies; Ensure; Environment; Eukaryota; Eukaryotic Cell; European; Event; Faculty; Fostering; Functional RNA; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic; genome-wide; Genomics; Goals; graduate student; Health; Hereditary Disease; Human; human disease; In Vitro; Individual; Informatics; insight; interest; International; Introns; Investigation; Knowledge; Laboratories; lectures; Length of Stay; man; meetings; Messenger RNA; Methods; MicroRNAs; Molecular; Molecular Biology; mouse model; mRNA Precursor; Mutation; new technology; Nuclear; Oral; Organism; Participant; Physiological; Play; Polyadenylation; Postdoctoral Fellow; posters; Process; Proteins; Published Comment; Publishing; Reaction; Reading; Recording of previous events; Recruitment Activity; Regulation; Regulatory Pathway; Request for Applications; Research; Research Institute; Research Personnel; Resources; RNA; RNA Interference; RNA Processing; RNA Splicing; Schedule; Science; Scientist; Seasons; Selection Criteria; Senior Scientist; Series; Shapes; Spinal Muscular Atrophy; Spliced Genes; Students; success; symposium; System; Therapeutic; Transcript; Transport Process; unpublished works; Woman; Work; Yeasts
Relevance: In genes of higher organisms from yeast to man, the information encoded in the DNA sequence is interrupted by non-coding regions called introns. An RNA copy of the gene has to be read off, cut and then spliced back together to remove the introns and produce a continuous "message" with the correct information to produce a protein. In many cases, the message can be cut and put back together in different combinations giving rise to proteins with different functions. This means that the number of different proteins in a cell can be much greater than the number of different genes. Mistakes in the splicing of the RNA cause serious problems as defective proteins are produced, and this sometimes happens as a consequence of genetic defects or disease. The molecular machinery that carries out this gene splicing is very complicated and has to be tightly regulated in the cell. Many scientists are studying how this messenger RNA processing occurs and is controlled, helped by the fact that the RNA splicing machinery is highly conserved between yeast and humans. This conference brings these scientists together to discuss their latest results. A key to the success of the conference is that the majority of oral presentations are given by graduate students, postdoctoral fellows and junior faculty chosen on the basis of scientific merit, ensuring that the conference showcases the latest developments which are often yet to be published. Participants come from academic centers, research institutes and industrial centers around the world to present and discuss their findings. Importantly, this application requests support for junior scientists who might not otherwise be able to attend to actively participate in the meeting
Project start date: 2011-06-03
Project end date: 2012-05-31
Budget start date: 3-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-10-071
1R13CA159737-01 (2011): $4000
CSHL 2011 CELLULAR BIOLOGY OF ADDICTION COURSE
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 2R13DA019791-04 from National Institute On Drug Abuse
Abstract: The primary objective of the proposed workshop is to provide an intense discussion of the fundamentals, state-of-the-art advances and major gaps in the cell and molecular biology of drug addiction. Targeted to both new and experienced investigators, the workshop will combine formal presentations and informal discussions to convey the merits and excitement of cellular and molecular approaches to drug addiction research. The workshop will stress the significance of this approach not only to the most costly neuropsychiatric disorder, addiction, but its applicability to learning, memory, neurochemical correlates of reward, perception and other disciplines of neuroscience. Research on the molecular and cellular changes that occur in response to drugs is critical for clarifying whether adaptive processes are diverse for each stage of addiction and for each drug of abuse. With the advent of genomics and proteomics, an extraordinary opportunity now exists to develop comprehensive models of neuroadaptative processes fundamental to addiction, craving, and relapse to drug use. The workshop will provide attendees with a spectrum of disciplines that can contribute to developing comprehensive models of each stage of drug addiction. The following disciplines will be presented 1.Overview of addiction. 2. Neuroadaptation to drugs of abuse. 3. Association between drug-induced neuroadaptation and behavioral manifestations 4. Genotype/susceptibility to drug addiction or drug response. 5. Tolerance, sensitization and adaptation at the cellular level. 6. New receptor targets. 7. Drug discovery and development to treat drug abuse how can cell biology contribute? This workshop will provide an integrated view of current and novel research on neuroadaptive responses to addiction, foster discussion on collaboration and integration and provide critical information needed to construct a model of addiction as a disease and novel molecular targets for biological treatments. Beyond the plane of scientific endeavor, such information is vital for formulating public policy and for enlightening the public on the neurobiological consequences of drug use and addiction. The proposed workshop will generate intense interest, open conduits for collaborations and novel routes to investigating the neurobiology of addiction. The workshop makes particular efforts to include a diverse group of participants, being particularly mindful of the inclusion of women, US minorities and a good balance of national and international scientists. The societal consequences of drug addiction and abuse are enormous. Over the past thirty years, behavioral studies have tended to dominate the field of addiction research. With the rapid growth in molecular neuroscience in the last two decades and the completion of the human genome in 2003, integration between behavioral or physiological responses and molecular genetic and cellular responses is increasingly possible and relevant. Amalgamation of these disciplines requires a profound appreciation for the strengths and weaknesses of each approach. The theme of the workshop is to provide attendees with the most up-to-date information on how tissues, cells and genes respond to drugs of abuse, within the framework of the behavioral effects of addictive drugs. This week-long residential workshop is targeted to both new and experienced investigators, and combines formal presentations and informal discussions to convey the merits and excitement of cellular and molecular approaches to drug addiction research. The workshop will stress the significance of this approach not only to the most costly neuropsychiatric disorder, addiction, but its applicability to learning, memory, and other disciplines of neuroscience. Research on the molecular and cellular changes that occur in response to drugs is critical for clarifying whether adaptive processes are diverse for each stage of addiction and for each drug of abuse. Ultimately, new treatments for these disorders are likely to emerge from a better understanding of the underlying processes that go awry during substance abuse and addiction
Keywords: addiction; Attention; Behavioral; Biological; Brain; Cells; Cellular biology; Collaborations; craving; design; Discipline; Disease; Disease model; Drug abuse; Drug Addiction; drug development; drug discovery; drug of abuse; Drug usage; Educational workshop; Effectiveness; Electronic Mail; Ensure; Equilibrium; Evaluation; experience; Fostering; Foundations; Genes; Genomics; Genotype; Human Genome; Individual; instructor; interdisciplinary approach; interest; International; Lead; Learning; lecturer; Measures; Memory; Mentors; Minority; Modeling; Molecular; Molecular Biology; Molecular Genetics; Molecular Target; neuroadaptation; Neurobiology; neurochemistry; neuropsychiatry; Neurosciences; New York; novel; Online Systems; Participant; Perception; Pharmaceutical Preparations; Physiological; Predisposition; present value; Process; programs; Proteomics; public health relevance; Public Policy; Publications; rapid growth; receptor; Relapse; Research; Research Personnel; response; Rewards; Route; satisfaction; Scientist; Spain; Staging; Stress; Structure; Students; Substance abuse problem; Substance Addiction; Suggestion; Surveys; Techniques; Tissues; Training; Woman
Relevance: Narrative The societal consequences of drug addiction and abuse are enormous. Over the past thirty years, behavioral studies have tended to dominate the field of addiction research. With the rapid growth in molecular neuroscience in the last two decades and the completion of the human genome in 2003, integration between behavioral or physiological responses and molecular genetic and cellular responses is increasingly possible and relevant. Amalgamation of these disciplines requires a profound appreciation for the strengths and weaknesses of each approach. The theme of the workshop is to provide attendees with the most up-to-date information on how tissues, cells and genes respond to drugs of abuse, within the framework of the behavioral effects of addictive drugs. This week-long residential workshop is targeted to both new and experienced investigators, and combines formal presentations and informal discussions to convey the merits and excitement of cellular and molecular approaches to drug addiction research. The workshop will stress the significance of this approach not only to the most costly neuropsychiatric disorder, addiction, but its applicability to learning, memory, and other disciplines of neuroscience. Research on the molecular and cellular changes that occur in response to drugs is critical for clarifying whether adaptive processes are diverse for each stage of addiction and for each drug of abuse. Ultimately, new treatments for these disorders are likely to emerge from a better understanding of the underlying processes that go awry during substance abuse and addiction
Project start date: 2005-07-01
Project end date: 2016-02-29
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-10-071
2R13DA019791-04 (2011): $79300
PODOCYTE-SPECIFIC HUMAN PLA2R TRANSGENIC MOUSE MODEL OF MEMBRANOUS NEPHROPATHY
J David
Boston Medical Centercity: Boston country: United States (us)
Grant 5R21AI090238-02 from National Institute Of Allergy And Infectious Diseases
Abstract: The primary goal of this exploratory proposal is to develop a tool to determine if circulating autoantibodies directed at the M-type phospholipase receptor (PLA2R) from patients with idiopathic membranous nephropathy (MN) are pathogenic. We have established that about 75% of patients with idiopathic MN have circulating anti-PLA2R autoantibodies whereas of the disease or normal controls has tested positive, thus indicating a high degree of specificity. The anti-PLA2R antibodies are IgG4, the subclass that predominates in idiopathic but not secondary MN, the antigen PLA2R is expressed on podocytes and colocalizes with IgG4 in the glomerular immune deposits of patients with MN, and the antibodies can be eluted from the glomerular immune deposits of MN renal biopsies. Moreover, the presence of circulating anti-PLA2R antibodies correlates with disease activity in MN. Although these findings strongly suggest that anti-PLA2R antibodies are responsible for, or at least contribute to disease development, proof of pathogenicity has yet to be established. Whereas simple transfer of the disease to laboratory animals with human anti-PLA2R would readily answer this question, human anti-PLA2R does not recognize PLA2R in the glomeruli of any small laboratory animal tested to date. Therefore, we plan to take advantage of both the lack of immunoreactive PLA2R in mouse glomeruli and the refinement of podocyte- specific transgenic techniques to develop a mouse model in which the human PLA2R is expressed in podocytes. To that end we propose the following specific aims Specific Aim 1 Produce a transgenic mouse utilizing the NPHS1 (nephrin) promoter to drive expression of human PLA2R exclusively in mouse podocytes. Specific Aim 2 Verify that human PLA2R is expressed on mouse podocytes in vivo. Specific Aim 3 a) Determine if injected human anti-PLA2R autoantibodies bind to human PLA2R and form immune deposits typical of MN in the transgenic mouse glomeruli. b) Establish if injected human anti-PLA2R antibodies are able to cause proteinuria in the human PLA2R transgenic mouse. Specific Aim 4 Determine if the human PLA2R transgenic mice develop anti-PLA2R antibodies and MN when actively immunized with human PLA2R. Successful development of this model will pave the way for studies to both establish the pathogenic role of human anti-PLA2R autoantibodies and define the mechanisms of injury. Having demonstrated that the M-type phospholipase receptor (PLA2R) is a specific target antigen of circulating autoantibodies in a high proportion of patients with idiopathic membranous nephropathy, we propose to develop a mouse model to determine if the antibodies are responsible for the clinical (proteinuria) and pathological manifestations of the disease. To that end, we will produce transgenic mice in which the human PLA2R is expressed on mouse podocytes, the location of the antigen in human kidneys
Keywords: Animal Testing; Antibodies; Antigen Targeting; Antigens; Autoantibodies; Binding (Molecular Function); Biopsy; Cavia; Cell Aging; Chinchilla (genus); Clinical; Complement; complement pathway; Deposition; Development; Disease; Eicosanoid Production; Epitopes; Future; Goals; Human; Idiopathic Membranous Nephropathy; IgG1; IgG4; Immune; Immunoglobulin G; in vivo; Injury; Kidney; Laboratory Animals; Ligand Binding; Location; Mediating; Membranous Glomerulonephritis; model development; Modeling; mouse model; Mus; nephrin; Oryctolagus cuniculus; Pathogenicity; Patients; Phospholipase; Play; podocyte; Positioning Attribute; Promotor (Genetics); Proteinuria; public health relevance; Rattus; receptor; Role; Serum; Specificity; System; Techniques; Testing; tool; Transgenic Mice; Transgenic Organisms
Relevance: Having demonstrated that the M-type phospholipase receptor (PLA2R) is a specific target antigen of circulating autoantibodies in a high proportion of patients with idiopathic membranous nephropathy, we propose to develop a mouse model to determine if the antibodies are responsible for the clinical (proteinuria) and pathological manifestations of the disease. To that end, we will produce transgenic mice in which the human PLA2R is expressed on mouse podocytes, the location of the antigen in human kidneys
Project start date: 2010-07-02
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-09-164
5R21AI090238-02 (2011): $209138
TOBACCO PACKAGING AND LABELING POLICIES: EXPANDING THE EVIDENCE ON NOVEL POLICIES
J David, Pi
Roswell Park Cancer Institute Corpcity: Buffalo country: United States (us)
Abstract: The primary goal of Project 2 is to build the evidence base in the area of tobacco packaging and labeling policies (Article 11 of the FCTC), in low, middle, and high income countries. Project 2 will complement the survey data described in Project 1 by collecting information directly from tobacco packages and by conducting experimental studies in "high-burden" ITC countries. Project 2 includes 4 sub-studies, each with a specific aim. Specific Aim 1 is to examine consumer perceptions of cigarette packaging design among youth and adults in 7 "high-burden" ITC countries China, India, Bangladesh, Mexico, Republic of Korea, the United States, and Germany. Parallel experimental studies will be conducted in each of the 7 countries to examine the impact of brand descriptors (e.g., light, mild, smooth, slims) and brand imagery (e.g., colors and design) on perceptions of risk and brand appeal. Overall, the study seeks to identify potentially misleading information on cigarette packages and to evaluate the potential impact of "plain" packaging regulations. In Specific Aim 2, parallel experimental studies will be conducted with youth adults in the same 7 high-burden ITC countries to evaluate the impact of health warnings on tobacco packages. The study will examine the impact of three types of health warnings on health beliefs, message recall, and smoking susceptibility text- only health warnings, pictorial warnings with graphic depictions of health effects, and pictorial warnings with personal testimonials. Specific Aim 3 will attempt to replicate the findings from the experimental studies in Aims 1 and 2 by integrating key measures from these studies into the population-based surveys in 7 low and middle-income ITC countries Thailand, Malaysia, Uruguay, India, China, Mexico, and Bangladesh. Specific Aim 4 will document industry practices in cigarette packaging and their association with tobacco labeling policies. This study will systematically collect and analyze package design and descriptors from 50 leading cigarette brands in each of 20 countries, including all ITC countries. Each of the studies in Project 2 will focus upon potential similarities and differences between low,middle, and high income countries in industry practice and the effectiveness of labeling policiesa primary theme of this P01 application
Keywords: Address; Area; Arousal; Bangladesh; China; Cigarette; Color; Complement; Country; Data; Descriptor; design; Effectiveness; Emotional; Epidemic; evidence base; Germany; Global Change; Goals; Health; health belief; Imagery; Income; India; Industry; Label; Light; Low income; Malaysia; Measures; Mediating; Mexico; novel; Perception; Policies; population based; Predisposition; public policy on tobacco; Regulation; research study; response; risk perception; Sampling; Smoking; South Korea; Surveys; Text; Thailand; Tobacco; tobacco control; trend; United States; Uruguay; Work; young adult
Relevance: RELEVANCE (Seeinstructions): Health warnings and tobacco labelling policies have been identified by the WHO as one of six critical policies to address the tobacco epidemic. This project will collect information in high, middle, and low income countries to address key gaps in the evidence, particularly in low and middle income countries
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
5P01CA138389-03_8480 (2011): $175791
REDUCING MEDICAL RISKS IN INDIVIDUALS WITH BIPOLAR DISORDER
J David, Thomas Detre Professor And Chair
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5R01MH081003-04 from National Institute Of Mental Health
Abstract: The primary aim of this project is to understand whether it is possible to reduce medical risk factors in adults with bipolar disorder and, in doing so, to improve psychiatric and functional outcomes. In response to PA-07- 211, Health Behavior Change in People with Mental Disorders, we will examine the role of behavioral risk factors and presumed behavioral mediators and moderators of health risk in individuals suffering from bipolar disorder. We will employ a multi-pronged, longitudinal treatment platform that integrates an innovative behavioral intervention with guideline based psychiatric care (Integrated Risk Reduction Intervention} IRRI) in order to target modifiable medical risk factors. IRRI is aimed at ameliorating sleep/wake and social rhythm disturbance and achieving modest weight reduction by increasing physical activity and improving nutrition and dietary habits, while at the same time providing optimal psychiatric care and medical monitoring. This will allow us to investigate the role that improvements in sleep/wake and social rhythm regularity, diet, and physical activity have in improving psychiatric and functional outcomes. In order to examine another set of possible pathways (i.e., that it is the amelioration of psychiatric symptoms that leads to improvements in physical health), we will contrast outcomes of subjects receiving IRRI with those of subjects receiving psychiatric care with medical monitoring (PCMM), which incorporates the same optimal psychiatric care plus monitoring of medical conditions. These aims will be achieved in a 24-month randomized treatment trial of 144 adult patients with bipolar disorder. We hypothesize that 1) IRRI compared with PCMM will result in a larger decrease in medical risk factors and greater improvement in sleep/wake and social rhythm disturbances; and 2) IRRI compared with PCMM will result in greater improvement in mood symptoms and functioning, particularly employment-related functioning. Our secondary hypotheses are that 1) The effect of IRRI on improved mood symptoms and functioning will be mediated by the effect of IRRI on medical risk factors and on sleep/wake and social rhythm disturbances; 2) the effect of IRRI on medical risk factors and sleep/wake and social rhythm disturbances will be mediated by patients´ regular participation and adherence; 3) High baseline medical risk factors will predict worse outcomes over the course of the study for patients receiving PCMM than for patients with similarly high baseline medical risk factors who receive IRRI; and 4) Higher within-subject variation in medical risk and sleep/wake and social rhythm disturbances over the course of the study will be associated with smaller decreases in medical risks, smaller improvements in sleep/wake and social rhythm disturbances, and smaller improvements in mood symptoms and functioning. Additionally, within-subject variation in medical risk parameters over the course of the study will be smaller in patients receiving IRRI than in those receiving PCMM. The public health consequences of the morbidity and mortality associated with bipolar disorder are enormous and have increased over the past decade, despite the introduction of new pharmacological interventions. If we could better assess the risk and protective factors underlying the accumulation of psychiatric and medical morbidity in patients with bipolar disorder, then early interventions could be designed to prevent the rapid accumulation of chronic disease and adverse prognoses leading to persistent poor functioning and disability. We hope to identify such risk and protective factors, intervene to reduce medical risk factors with the expectation that psychiatric outcomes will be substantially improved, and thus provide a framework for further efforts at secondary prevention
Keywords: Address; Adherence (attribute); Adult; Affect; Alcohol or Other Drugs use; Area; base; Behavior; Behavior Therapy; Behavioral; Bipolar Disorder; Blood Vessels; Body Weight decreased; Chronic; Chronic Disease; Compliance behavior; depressive symptoms; design; Diet; Diet Habits; disability; Disease; Disease remission; Early treatment; Employment; Endocrine; epidemiologic data; evidence based guidelines; expectation; Functional disorder; functional outcomes; Guidelines; Health; Health behavior change; Immunologics; improved; Individual; innovation; Intervention; intervention effect; Lead; Left; Life Style; Mediating; Mediator of activation protein; Medical; Mental Depression; Mental disorders; Metabolic syndrome; Modeling; Monitor; Mood Disorders; Moods; Morbidity - disease rate; Mortality Vital Statistics; Neurophysiology - biologic function; NIH Program Announcements; nutrition; Outcome; outcome forecast; Pathway interactions; Patient Monitoring; patient population; Patients; Physical activity; physical conditioning; prevent; programs; Prospective Studies; Psychiatric therapeutic procedure; public health medicine (field); public health relevance; Randomized; Recurrence; response; Risk; Risk Factors; Risk Reduction; Role; Secondary Prevention; Sleep; Smoking; social; Suicide; Symptoms; Testing; Time; Translational Research; Treatment outcome; treatment trial; Variant; Work
Project start date: 2008-08-15
Project end date: 2013-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-07-211
5R01MH081003-04 (2011): $630363
2011 COLD SPRING HARBOR CONFERENCE ON NEUROBIOLOGY OF DROSOPHILA
J David, Executive Director, Meetings And Courses
Cold Spring Harbor Laboratorycity: Cold Spring Harbor country: United States (us)
Grant 1R13NS076086-01 from National Institute Of Neurological Disorders And Stroke
Abstract: The planned conference on Neurobiology of Drosophila will convene a group of junior and senior scientists to discuss the latest advances in the neurobiology research that are being made in the highly successful model system the fruit fly Drosophila melanogaster. This proposal seeks support for the 2011 conference, which is the 14th biennial international meeting in this series. This meeting focuses on advances made using the combined power of genetics, molecular biology, biochemistry, cell biology, electrophysiology, imaging, and behavioral analysis to address fundamental issues in neurobiology. The topics covered will range from i) Sensory systems ii) Behavior, brain function & its evolution; iii) Neurological disease models & cellular mechanisms; iv) Neural circuits - development, evolution and function; v) Synaptic transmission, development and plasticity; vi) Neural development & evolution; and vii) Technological innovations & applications. By vote of the previous participants, the meeting will remain of moderate size (430 participants) and will have no parallel sessions so as to facilitate discussion, exchange of ideas and techniques, and to promote new collaborations in this rapidly-evolving field. All applicants will be encouraged to submit an and the majority of participants will present a talk or poster. Speakers will be chosen by session leaders and meeting organizers from the most timely and interesting s submitted a few months in advance of the conference this will ensure that late-breaking science is covered in all of the talks. In the event that the conference is oversubscribed, participants will be chosen to include at least one representative from each participating laboratory. This conference has an excellent track record for highlighting the work of younger investigators and women. To encourage participation by junior investigators, a special lecture is presented by a graduate student who has written the best Ph.D. thesis since the previous meeting. This Neurobiology of Drosophila conference focuses on advances made using the combined power of genetics, molecular biology, biochemistry, cell biology, electrophysiology, imaging, and behavioral analysis to address fundamental issues in neurobiology using the model organism Drosophila melanogaster. A driving principle of the entire field is that the lowly fruit fly, while seeming to have little in common with humans, shares many of the same genes, molecules and neural wiring -albeit far simpler - with vertebrate animals. Building on one hundred years of research in fly genetics, the fly neurobiology field is a burgeoning field of endeavor with many laboratories around the world making significant advances in the field. Understanding how these systems work in Drosophila has enormous relevance to our efforts to elucidate how these things function in higher animals, with consequences including how we think about neurological disease, brain evolution and other major themes of research in humans. Many of the research techniques that are developed for use in the fly have considerable utility in research into nervous system function in vertebrates and mammals. The aim of the conference is to provide a forum to bring together a diverse group of scientists working in different areas of the field of fly neurobiology. The meeting will be of moderate size and will have no parallel sessions so as to facilitate discussion, exchange of ideas and techniques, and to promote new collaborations in this rapidly evolving field. The scientific organizers are elected by vote of the previous participants, thus ensuring a fresh perspective on the scientific exchange. All applicants are encouraged to submit an and the majority of participants will present a talk or poster. Speakers will be chosen by session leaders and the meeting organizers from the most timely and interesting s submitted a few months in advance of the conference, ensuring that late-breaking science is covered in all of the talks
Keywords: ing; Accounting; Address; Animal Model; Animals; Area; Asians; Automobile Driving; base; Behavior; Behavioral; Biochemistry; Biological; Biological Models; Brain; Brain Diseases; Businesses; Cells; Cellular biology; Circadian Rhythms; Collaborations; Complex; Data; Development; Disease; Disease model; Doctor of Philosophy; Drosophila genome; Drosophila genus; Drosophila melanogaster; Electrophysiology (science); Ensure; Environment; European; Event; Evolution; Faculty; fly; Fostering; Future; Genes; Genetic; Goals; graduate student; Health; Human; Image; insight; interest; International; Laboratories; laboratory facility; lectures; Length of Stay; Maintenance; Mammals; meetings; Molecular; Molecular Biology; nervous system disorder; Nervous System Physiology; Nervous system structure; neural circuit; Neurobiology; neurodevelopment; Neurons; Neurosciences; Oral; Participant; Physiological; Postdoctoral Fellow; posters; Published Comment; Questionnaires; relating to nervous system; Research; Research Institute; Research Personnel; Research Technics; Schedule; Science; Scientist; Senior Scientist; sensory system; Series; success; symposium; Synaptic Transmission; System; Techniques; technological innovation; Transcriptional Regulation; translational study; unpublished works; Vertebrates; Voting; Woman; Work; Writing
Relevance: This Neurobiology of Drosophila conference focuses on advances made using the combined power of genetics, molecular biology, biochemistry, cell biology, electrophysiology, imaging, and behavioral analysis to address fundamental issues in neurobiology using the model organism Drosophila melanogaster. A driving principle of the entire field is that the lowly fruit fly, while seeming to have little in common with humans, shares many of the same genes, molecules and neural wiring -albeit far simpler - with vertebrate animals. Building on one hundred years of research in fly genetics, the fly neurobiology field is a burgeoning field of endeavor with many laboratories around the world making significant advances in the field. Understanding how these systems work in Drosophila has enormous relevance to our efforts to elucidate how these things function in higher animals, with consequences including how we think about neurological disease, brain evolution and other major themes of research in humans. Many of the research techniques that are developed for use in the fly have considerable utility in research into nervous system function in vertebrates and mammals. The aim of the conference is to provide a forum to bring together a diverse group of scientists working in different areas of the field of fly neurobiology. The meeting will be of moderate size and will have no parallel sessions so as to facilitate discussion, exchange of ideas and techniques, and to promote new collaborations in this rapidly evolving field. The scientific organizers are elected by vote of the previous participants, thus ensuring a fresh perspective on the scientific exchange. All applicants are encouraged to submit an and the majority of participants will present a talk or poster. Speakers will be chosen by session leaders and the meeting organizers from the most timely and interesting s submitted a few months in advance of the conference, ensuring that late-breaking science is covered in all of the talks
Project start date: 2011-09-01
Project end date: 2012-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-10-071
1R13NS076086-01 (2011): $15000
DETERMINING EPIGENETIC TARGETS OF MILK THISTLE EXTRACT IN PROSTATE
J David, Professor And Chair
North Carolina Central Universitycity: Durham country: United States (us)
Grant 7R21CA135503-03 from National Cancer Institute
Abstract: The overall goal of the proposed research is to discover allelic targets for epigenetic regulation by milk thistle-derived compounds. Extracts of milk thistle (Silybum marianum L. Gaertn) have been investigated as experimental preventive and therapeutic agents against a variety of epithelial cancers including prostate cancer. While the precise direct or indirect molecular targets of the active compounds in this extracts have not been identified, we show preliminary evidence that the partially purified milk thistle extract silibinin affects the regulation and activity of the protein EZH2 in a variety of prostate tumor cell liners. EZH2 is a protein methyltransferase that can alter chromatin structure and lead to recruitment of DNA methyltransferases to gene promoters. It is also been characterized as a biomarker of the more aggressive prostate and breast cancers in humans. Given the effects of silibinin on a known modifier of epigenetic events, we hypothesize that the tumor preventative properties of milk thistle may proceed through epigenetic mechanisms. The proposed work takes an exploratory genome-wide approach to uncovering epigenetic targets of silibinin by developing a two part screening system. In the first screen, alleles that show high expression variation in the absence of genetic heterogeneity are determined by examining protein expression in normal human prostate tissue from unrelated individuals undergoing prostatectomies for non-prostate cancer related reasons. In the second screen, alleles that show differential response to repeated dosing of silibinin are identified using a prostate tumor cells line. The hypothesis being tested is that alleles the show high expression level heterogeneity in a normal tissue, that are also affected by repeated exposure to silibinin are metastable epialleles; a.k.a, alleles most susceptible to epigenetic regulation in normal tissue. By identifying these alleles and understanding their epigenetic heterogeneity in a normal tissue, we advance our understanding of how dietary factors such as silibinin exert their effects on prostate tissue over the long term. The overall goal of the proposed research is to discover metastable epialleles that are influenced by milk thistle extracts. Extracts of milk thistle (Silybum marianum L. Gaertn) have been investigated as experimental preventive and therapeutic agents against a variety of epithelial cancers including prostate cancer. By identifying these alleles and understanding their epigenetic heterogeneity in a normal tissue, we advance our understanding of how dietary supplements such as silibinin exert their effects on prostate tissue over the long term
Relevance: The overall goal of the proposed research is to discover metastable epialleles that are influenced by milk thistle extracts. Extracts of milk thistle (Silybum marianum L. Gaertn) have been investigated as experimental preventive and therapeutic agents against a variety of epithelial cancers including prostate cancer. By identifying these alleles and understanding their epigenetic heterogeneity in a normal tissue, we advance our understanding of how dietary supplements such as silibinin exert their effects on prostate tissue over the long term
Project start date: 2009-05-10
Project end date: 2012-04-30
Budget start date: 15-SEP-2011
Budget end date: 30-APR-2012
PFA/PA: PA-06-413
7R21CA135503-03 (2010): $92396
INTERDISCIPLINARY TRAINING IN GENOME ENGINEERING (COMPONENT 10 OF 11)TL1
J David, Head, Section Of Immunology
Seattle Children´s Hospitalcity: Seattle country: United States (us)
Grant 5TL1HL092556-05 from Office Of The Director, National Institutes Of Health
Abstract: The overall goal of the NGEC is the application of genome engineering to hematopoietic stem cells for the purpose of gene repair. Achieving this goal will require work among the seven NGEC scientific components to be tightly integrated. A secondary aim of the NGEC is to promote the development of genome engineering as a scientific discipline. This T90/R90 component will function to promote both of these objectives by supporting interdisciplinary training of post-doctoral level individuals in genome engineering- related fields. Selected individuals will carry out projects spanning two or more disciplines with co-mentoring by two principal investigators with complementary expertise. Support is requested for one position in year one, three positions in year 2, and four positions in years 3-5. Fellows will be chosen on the basis of the interdisciplinary nature of their project and its relationship to genome engineering. The period of training will be up to two years, and training will involve active research and associated laboratory meetings, NGEC group meetings (see component 1), presentation of their data at the annual Workshop for Genome Engineering (see component 9), and travel to an outside meeting each year. Options are also provided for additional interdisciplinary experiences including didactic coursework, a biotechnology externship, the many interdisciplinary symposia and workshops which take place among NGEC institutions, and graduate level teaching
Keywords: base; Biotechnology; Data; Development; Discipline; Educational process of instructing; Educational workshop; experience; gene repair; Genome engineering; Goals; Group Meetings; Hematopoietic stem cells; Individual; Institution; Laboratories; meetings; Mentors; Nature; Positioning Attribute; post-doctoral training; Postdoctoral Fellow; Principal Investigator; symposium; Training; Travel; Work
Project start date: 2007-09-30
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: RFA-RM-06-008
5TL1HL092556-05 (2011): $82876
INTERDISCIPLINARY TRAINING IN GENOME ENGINEERING(COMPONENT 10 OF 11) RL9
J David, Head, Section Of Immunology
Seattle Children´s Hospitalcity: Seattle country: United States (us)
Grant 5RL9HL092555-05 from Office Of The Director, National Institutes Of Health
Abstract: The overall goal of the NGEC is the application of genome engineering to hematopoietic stem cells for the purpose of gene repair. Achieving this goal will require work among the seven NGEC scientific components to be tightly integrated. A secondary aim of the NGEC is to promote the development of genome engineering as a scientific discipline. This T90/R90 component will function to promote both of these objectives by supporting interdisciplinary training of post-doctoral level individuals in genome engineeringrelated fields. f Selected individuals will carry out projects spanning two or more disciplines with co-mentoring by two principal investigators with complementary expertise. Support is requested for one position in year one, three positions in year 2, and four positions in years 3-5. Fellows will be chosen on the basis of the interdisciplinary nature of their project and its relationship to genome engineering. The period of training will be up to two years, and training will involve active research and associated laboratory meetings, NGEC group meetings (see component 1), presentation of their data at the annual Workshop for Genome Engineering (see component 9), and travel to an outside meeting each year. Options are also provided for additional interdisciplinary experiences including didactic coursework, a biotechnology externship, the many interdisciplinary symposia and workshops which take place among NGEC institutions, and graduate level teaching
Keywords: Area; base; Biotechnology; Data; Development; Discipline; Educational process of instructing; Educational workshop; experience; Future; gene repair; Genome; Genome engineering; Goals; Group Meetings; Hematopoietic stem cells; Individual; Institution; Laboratories; meetings; Mentors; Methods; Nature; Positioning Attribute; post-doctoral training; Postdoctoral Fellow; Principal Investigator; programs; Research Personnel; symposium; System; Training; Travel; Ursidae Family; Work
Project start date: 2007-09-30
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: RFA-RM-06-008
5RL9HL092555-05 (2011): $132116
MOLECULAR CONTROL OF NEURAL CELL FATE DETERMINATION
J David, Professor
California Institute Of Technologycity: Pasadena country: United States (us)
Grant 5R01NS023476-23 from National Institute Of Neurological Disorders And Stroke
Abstract: The objective of this proposal is to understand the cellular and molecular mechanisms that control a fundamental event in neural development the switch from neurogenesis to gliogenesis. An understanding of this process is essential for applying neural stern and progenitor cell biology to the treatment of neurological disease. This switch will be studied in a specific population of spinal cord precursors that sequentially generate motoneurons (MNs) and oligodendracytes (oligos). These precursors can be prospectively isolated using fluorescence-activated cell sorting (FACS), by means of a GFP reporter expressed from the Olig2 locus, which encodes a transcription factor required for both MN and oligo differentiation. Using these isolated cells, we will address the following specific aims I). We will test whether MNs and oligos develop from a multipotential, self-renewing stem cell in the ventricular zone (VZ) of the spinal cord, as is widely assumed, or rather from progenitors that undergo irreversible restrictions in developmental competence. We will investigate this by using a newly developed technique for direct transplantation of freshly isolated Olig2-expressing progenitors into the chick spinal cord, without any ex vivo expansion (which may perturb the properties of the cells). Using this approach, we will perform heterochronic transplantation experiments to test the self-renewal anc developmental capacities of Olig2-i- cells at different stages during the MNgoligo transition. II). We will test the hypothesis that changes in gene expression in Olig2+ progenitors during the neuron-to-glial switch reflect the regulation of several distinct subclasses of genes, each with different kinetics of activation and repression. This hypothesis will be tested by using oligonucleotide microarrays to perform gene expression profiling (GEP) experiments on acutely isolated Olig2+ progenitors from different stages of spinal cord development. This GEP analysis should also identify a) markers useful in clarifying lineage relationships between Olig2+ progenitors of MNs and oligos; and b) candidate regulatory genes for functional analysis. Ill) We will tost the hypothesis that there is a "temporal code" of transcription factors that controls the MN->oligo switch, by performing gain-of-function (GOF) and loss-of-function (LOF) genetic manipulations of candidate regulatory genes identified in the GEP temporal analysis (Aim II). Electroporation of chick spinal cord will be used as a rapid in vivo assay for such functional manipulations, employing expression of full-length cDNAs, and independently validated shRNAi (small hairpin RNAi) constructs, for GOF and LOF experiments, respectively. The embryos will be analyzed uning an extensive battery of molecular markers for various classes of neurons (including MNs and interneurons), oligodendrocytes, and newly validated markers of astrocytes and their progenitors. Candidates for which strong functional data is obtained from the chick system will be further validated by generating constitutive or conditional knockouts in the mouse. IV) We will test the hypothesis that targets of OLIG2, which functions as a transcriptional repressor, include a) repressers of oligo differentiation; and b) activators of astrocyte differentiation. Candidate targets of OLIG2 will be identified by performing comparative GEP analysis of isolated Olig2-GFP+ cells from Olig1/2+/- and Olig2-/- spinal cord, at several developmental stages. A series of analytic algorithms will be used to filter the data to obtain a list of transcription factors that are de-repressed in the absence of OLIG2 function. These candidates will be further validated and prioritized by real-time RT-PCR and in situ hybridization. Top candidates will then be functionally analyzed by GOF and LOF manipulations in chick embryos
Keywords: Address; Algorithms; Astrocytes; base; Biological Assay; Candidate Disease Gene; Categories; Cell Communication; Cells; Cellular biology; Chick Embryo; Classification; Code; comparative; Competence; Data; Development; Electroporation; Embryo; Event; Exhibits; Fluorescence-Activated Cell Sorting; gain of function; Gene Expression; Gene Expression Profiling; gene interaction; Genes; genetic manipulation; gliogenesis; In Situ Hybridization; In Vitro; in vivo; Interneurons; Kinetics; Knock-out; Length; loss of function; Maps; Methods; Molecular; molecular marker; Motor Neurons; Mus; mutant; nervous system disorder; neurodevelopment; neurogenesis; Neuroglia; Neurons; Oligodendroglia; Oligonucleotide Microarrays; Oligonucleotides; Pattern; Play; Population; Process; progenitor; Property; Regulation; Regulator Genes; relating to nervous system; Reporter; Repression; research study; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Role; self-renewal; Series; Spinal Cord; Staging; stem; Stem cells; System; Techniques; Testing; Time; transcription factor; Transcription Repressor/Corepressor; Transplantation; Ventricular
Project start date: 1986-09-01
Project end date: 2011-04-30
Budget start date: 1-MAY-2009
Budget end date: 30-APR-2011
5R01NS023476-23 (2009): $342108
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