MAMMALIAN FOREGUT AND LIVER DEVELOPMENT

M James
Children´s Hospital Medical Center Cincicity: Cincinnati    country: United States (us)

Grant 5R01DK080823-04 from National Institute Of Diabetes And Digestive And Kidney Diseases

Keywords: Anterior; Bile fluid; Blood Proteins; Cardiac; Cell Adhesion; Cell Culture Techniques; cell type; Cells; Data; Defect; Development; Disease; Dose; Drug Metabolic Detoxication; Embryo; Embryonic Development; embryonic stem cell; Endocrine; Endoderm; Event; Exhibits; Exocrine Glands; Fibroblast Growth Factor; Gastrula; Gene Expression; Generations; Genetic; Genetic Programming; Glucose; Glycogen; Goals; Hepatic; Hepatocyte; Hindgut; Human; human disease; human embryonic stem cell; In Vitro; insight; Intestines; Knock-out; Knowledge; Lateral; Life; Ligands; Link; Liver; Liver diseases; liver transplantation; Mammals; Mediating; Mesoderm; Methods; Modeling; Molecular; monolayer; Morphogenesis; Mus; mutant; Operating System; Organ; Pathway interactions; Pattern; Phenotype; Primitive foregut structure; progenitor; Publishing; receptor; Reporting; Repression; Research; research study; response; Role; Signal Transduction; Site; Somites; Source; Staging; Testing; Time; Tissue Transplantation; Tissues; Translating; Undifferentiated; United States National Institutes of Health; Xenopus

Relevance: Narrative The liver is a vital organ providing many essential functions and numerous diseases are so life threatening that liver transplantation is the only option. The differentiation of liver cells from embryonic stem cells is a potentially renewable source of tissue for transplantation. The goal of this proposal is to elucidate the genetic programs controlling embryonic liver development in mice. We will then use this information to recapitulating the key embryonic events in human embryonic stem cells to more effectively generate liver tissue in culture

Project start date: 2009-01-01

Project end date: 2012-12-31

Budget start date: 1-JAN-2012

Budget end date: 31-DEC-2012

5R01DK080823-04 (2012): $331268

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MAMMALIAN FOREGUT AND LIVER DEVELOPMENT

M James, Assistant Professor
Children´s Hospital Med Ctr (cincinnati)city: Cincinnati    country: United States (us)

Grant 5R01DK080823-03 from National Institute Of Diabetes And Digestive And Kidney Diseases

Abstract: The long-term goal of this proposal is to elucidate the molecular mechanism controlling mammalian foregut and early liver development. The largest exocrine gland in the body the liver produces bile and is the primary site for detoxification. It also performs important endocrine functions by secreting homeostatic blood proteins and regulating glucose levels through glycogen storage. A recent trans-NIH report "Action Plan for Liver Disease Research (2004)" recognized that a better understanding of embryonic liver development would provide important insights into human liver disease and promote our ability to harness embryonic stem cells as a renewable source of tissue for transplantation. The mouse embryonic liver is induced from the ventral foregut endoderm by FGF signals from the cardiac mesoderm. While we increasingly understand the genetic pathways regulating proliferation and differentiation of hepatoblasts after the liver bud has formed, the earlier events linking endoderm patterning to hepatic specification are less clear. In Xenopus we recently determined that differential Wnt/beta-catenin signaling regulates endoderm fates. Our data supports a model where during gastrula and early somite stages secreted Wnt- antagonists in the anterior endoderm establish foregut identity and initiate a molecular cascade leading to liver development. In contrast, the posterior endoderm has high beta-catenin activity, due to Wnt ligands secreted from the lateral/axial mesoderm, which represses foregut fate and promotes intestinal development. We propose to test this hypothesis using mouse genetics and embryonic explants to characterize the underlying molecular mechanism. Moreover, we will investigate whether analogous pathways are important for liver development in humans using endoderm cultures derived from human embryonic stem cells. The results of this proposal will directly impact efforts to generate therapeutically useful endoderm tissue for the treatment of liver disease in humans. Aim 1. Determine if repression of beta-catenin activity in the anterior endoderm is required for foregut and liver development using mouse genetics. Aim 2. Define when beta-catenin needs to be repressed and examine how the temporally distinct Wnt and FGF pathways interact during hepatic development, using mouse embryonic explant cultures. Aim 3. Investigate the role of Wnt signaling in promoting human foregut and liver lineages from HESCs. PUBLIC HEALTH RELEVANCE. The liver is a vital organ providing many essential functions and numerous diseases are so life threatening that liver transplantation is the only option. The differentiation of liver cells from embryonic stem cells is a potentially renewable source of tissue for transplantation. The goal of this proposal is to elucidate the genetic programs controlling embryonic liver development in mice. We will then use this information to recapitulating the key embryonic events in human embryonic stem cells to more effectively generate liver tissue in culture

Keywords: Anaplastic; Anterior; beta catenin; Bile; Bile fluid; Bile Juice; biological signal transduction; Blood Proteins; body system, hepatic; Body Tissues; bowel; Cadherin-Associated Protein, Beta; Cadherin-Associated Protein, Beta 1 (88kD); Cardiac; Catenin, Beta-1; Cell Adhesion; Cell Communication and Signaling; Cell Culture Techniques; Cell Signaling; cell type; Cells; Cellular Adhesion; CTNNB1; CUL-2; D-Glucose; Data; Defect; detoxification; Development; Dextrose; Disease; disease/disorder; Disorder; DNA Synthesis Factor; Dose; Drug Metabolic Detoxication; ECGF; Embryo; Embryo Development; Embryogenesis; Embryonic; Embryonic Development; embryonic stem cell; Endocrine; Endoderm; Endothelial Cell Growth Factor; ES cell; Event; Exhibits; Exocrine Glands; experiment; experimental research; experimental study; FGF; Fibroblast Growth Factor; Fibroblast Growth Regulatory Factor; Foregut; Gastrula; Gene Expression; Generations; Genetic; Genetic Algorithm; Genetic Programming; Glucose; Glycogen; Goals; Grafting, Liver; HBGF; Health; Hepatic; Hepatic Cells; Hepatic Disorder; Hepatic Parenchymal Cell; Hepatocyte; hepatopathy; hESC; Hindgut; Human; human disease; human embryonic stem cell; human ES cell; human ESC; Human, General; In Vitro; insight; Intestinal; Intestines; Intracellular Communication and Signaling; Knock-out; Knockout; Knowledge; language translation; Lateral; Life; Ligands; Link; Liver; Liver Cells; Liver diseases; liver disorder; Liver Transplant; liver transplantation; Mammalia; Mammals; Mammals, General; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Mesoderm; Metabolic Detoxication, Drug; Metabolic Detoxification, Drug; Metabolic Drug Detoxications; Metabolism of Toxic Agents; Methods; Mice; Modeling; Molecular; monolayer; Morphogenesis; Murine; Mus; mutant; National Institutes of Health; National Institutes of Health (U.S.); NIH; Operating System; Organ; organ system, hepatic; pathway; Pathway interactions; Pattern; Phenotype; Primitive foregut structure; PRO2286; progenitor; Publishing; receptor; Receptor Protein; Reporting; Repression; Research; research study; response; Role; Signal Transduction; Signal Transduction Systems; Signaling; Site; social role; Somites; Source; Staging; stem cell of embryonic origin; Testing; Time; Tissue Transplantation; Tissues; Translating; Translatings; Transplantation of liver; Transplantation, Hepatic; Undifferentiated; United States National Institutes of Health; Xenopus

Relevance: Narrative The liver is a vital organ providing many essential functions and numerous diseases are so life threatening that liver transplantation is the only option. The differentiation of liver cells from embryonic stem cells is a potentially renewable source of tissue for transplantation. The goal of this proposal is to elucidate the genetic programs controlling embryonic liver development in mice. We will then use this information to recapitulating the key embryonic events in human embryonic stem cells to more effectively generate liver tissue in culture

Project start date: 2009-01-01

Project end date: 2012-12-31

Budget start date: 1-JAN-2011

Budget end date: 31-DEC-2011

PFA/PA: PA-07-026

5R01DK080823-03 (2011): $337895

A MOUSE MODEL FOR COMPLEX HUMAN DISEASES

M James, Professor Of Anatomy
Washington Universitycity: Saint Louis    country: United States (us)

Grant 5R24RR015116-07 from National Center For Research Resources

Abstract: We propose to continue the development and maintenance of the LGXSM Recombinant Inbred (Rl) strain set and the associated Advanced Intercross (Al) Line formed by the intercross of inbred mouse strains LG/J and SM/J. The RIs are a tool for preliminary gene mapping, after which the Al Line can be used for finemapping quantitative trait loci to sub-cM regions. Under prior support we have developed a set of 18 Rl lines formed from the intercross of LG/J and SM/J and brought the Al line to the 32nd generation of random mating. The Rl lines have been scored for over 500 polymorphic microsatellite markers and for 4290 polymorphic SNPs, a polymorphic SNPs every 600 kb. Embryos have been cryopreserved for each Rl lines. These lines and other populations formed from the intercross of LG/J and SM/J have allowed the mapping of a large array of phenotypes relevant to disease processes of interest across the NIH Institutes. Here, we propose to enhance the Rl strain set by developing another 50 Rl strains using the present Al line as the source population. The addition of these strains will greatly increase the power of the Rl strain set in gene mapping studies. The new strains will be even more useful than the earlier set in that they will have accumulated 16 times the amount of recombination expected in Rl lines formed from a F2 generation. Furthermore, we will be able to use our prior breeding experience to increase strain survival to 50% in the production of new strains by selecting against specific loci that, singly and in combination, result in strain loss. The new strains will be genetically characterized for 500 microsatellites and 4290 SNPs and archived by embryonic cryopreservation. Strain genotypes and phenotypes will be made available through our web site, the Mouse Phenome Project, and the WebQTL database. In our earlier breeding, we discovered strong selection against the agouti alleles carried by SM/J, especially in the presence of the wild-type tyrosinase allele from that strain. We will examine the genetic basis for strain loss by sequencing the structural genes at these two loci and closely linked loci. We will produce 2ble congenic strains that are SM/J across the genome, except at the agouti and tyrosinase regions. Backcrossing with SM/J will be used to narrow the QTL interval for strain loss and to produce a new strain, carrying the unusual phenotypes of SM/J without requiring enforced heterozygosity at the agouti locus

Keywords: A Mouse; Affect; After Care; After-Treatment; Aftercare; Alleles; allelic variant; Allelomorphs; Anatomic; Anatomical Sciences; anatomy; Anatomy; Animals; Archives; Back; Backcrossings; base; Basic Research; Basic Science; Biological Models; Birth; Breeding; BRO; Brothers; Caring; Characteristics; Chromosome 2; Chromosome 7; Chromosome Mapping; Chromosomes, Human, Pair 2; Chromosomes, Human, Pair 7; clinical data repository; clinical data warehouse; cold preservation; cold storage; Color; Communities; Complement; Complement Proteins; Complex; congenic; Congenic Strain; cost; Cresolase; Cryofixation; Cryopreservation; Data; Data Banks; Data Bases; data repository; Databank, Electronic; Databanks; Database, Electronic; Databases; Decision Making; density; Development; Disease; disease/disorder; Disorder; DNA Recombination; DNA recombination (naturally occurring); Dopa Oxidase; Dorsum; EC 1.14.18.1; Embryo; Embryonic; Epistasis; Epistasis, Genetic; Epistatic Deviation; experience; Fecundability; Fecundity; Fertility; Funding; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; Gene variant; gene x gene interaction; Generations; Genetic; Genetic Diversity; genetic epistases; Genetic Epistasis; genetic mapping; Genetic Models; Genetic Recombination; Genetic Variation; Genetics, Gene Mapping; Genome; Genome Mappings; Genomics; Genotype; human disease; Inbred Mouse; Inbred Strain; Inbred Strains Mice; Inbreeding; Institutes; Interaction Deviation; interest; Investigators; LB24-AB; LG/J Mouse; Life; Link; Linkage Mapping; Location; Maintenance; Maintenances; Mammals, Mice; Maps; mate; Mice; Mice, Mutant Strains; Microsatellite Markers; Microsatellite Repeats; Microsatellites; Model System; Models, Biologic; Monophenol Monooxygenase; monophenol oxidase; Monophenol, L-dopa[{..}]oxygen oxidoreductase; mouse model; mouse mutant; Murine; Mus; Mutant Strains Mice; National Institutes of Health; National Institutes of Health (U.S.); neurobiological; Neurobiology; NIH; Partner in relationship; Parturition; Phase; Phenol Oxidase; Phenoloxidase; phenome; Phenotype; Polymorphic marker; Polymorphic Microsatellite Marker; Population; population based; Position; Positioning Attribute; Preparation; Process; Production; Publications; QTL; Quantitative Trait Loci; R01 Mechanism; R01 Program; Recombinants; Recombination; Recombination, Genetic; Recovery; Reimplantation; relational database; Replantation; repository; Research; Research Grants; Research Personnel; Research Project Grants; Research Projects; Research Projects, R-Series; Research Resources; Researchers; Resources; RPG; Science of Anatomy; Scientific Publication; Series; Shadowing; Shadowing (Histology); SIS; Sister; SK29-AB; Source; Staging; Structural Genes; study characteristics; Surgical Replantation; Testing; Time; tool; trait; Tumor Rejection Antigen AB; TYR; Tyrosinase; United States National Institutes of Health; Universities; Variation (Genetics); Washington; web site

Project start date: 2001-04-01

Project end date: 2011-05-31

Budget start date: 1-JUN-2009

Budget end date: 31-MAY-2011

5R24RR015116-07 (2009): $362790

INVESTIGATION OF DUF1220 DOMAINS IN HUMAN BRAIN FUNCTION AND DISEASE

M James, Professor
University Of Colorado Denvercity: Aurora    country: United States (us)

Grant 5R01MH081203-03 from National Institute Of Mental Health

Abstract: We have established that human genome sequences encoding a novel protein domain, DUF1220, are highly amplified in the human lineage (>200 copies vs. 1 in mouse/rat) and may be important in human-specific cognitive function. The majority of DUF1220 domains are located at 1q21.1, one of the most complex regions of the human genome and filled with gaps and segmental duplications. Copy number variations (CNVs) in the 1q21.1 region have now been implicated in numerous diseases associated with cognitive dysfunction, e.g. autism, mental retardation, schizophrenia, microcephaly and macrocephly. These findings may be indicative of a novel recurrent rearrangement and reflect a new cognition-related syndrome specific for the 1q21.1 region. In order to more precisely identify the CNV boundaries and causal disease genes in these patients, a haploid BAC library will be used to generate a finished sequence map of the region. Sequencing of 1q21.1 BACs from a Hydatiform (haploid) mole library will be carried out in collaboration with Dr. Rick Wilson at the Washington Univ. at St. Louis Genome Center to generate a single haplotype path across the region. The finished 1q21.1 sequence will be used for fine mapping of already identified disease-associated CNVs for autism, mental retardation, microcephaly and macrocephaly, through collaborations with the laboratories of Drs. Evan Eichler and James Lupski. High-density custom tiling arrays will be generated for the finished 1q21.1 region and used for array CGH to fine map CNV breakpoints in these patients and identify candidate genes. In addition the role of DUF1220 domain copy number in autism will be investigated by QPCR analysis of individuals with autism using DUF1220-specific primers. To investigate the function of DUF1220 domains in a living mammal, DUF1220-minus mice we have generated (the first animal model for DUF1220 function) will be subjected to behavioral testing to assess the affect of DUF1220 domain loss on learning and memory. In addition to providing the scientific community with the most complete genome map for this complex genomic region, these studies should lead to a better understanding of which genes in the 1q21.1 region, including those encoding DUF1220 domains, underlie the specific diseases of cognition that have been shown to be associated with CNVs in this region. In addition, the behavioral studies using DUF1220-minus mice should generate the first insights into the possible cognitive function of these domains in a living mammal

Keywords: Affect; Animal Model; Autistic Disorder; base; behavior test; Behavioral; Biological Markers; Brain; Candidate Disease Gene; Cognition; Cognition Disorders; Cognitive; cognitive function; Collaborations; Communities; Complex; Copy Number Polymorphism; Custom; Data; density; design; disability; Disease; DNA; DNA Sequence Rearrangement; Exhibits; Genes; Genome; Genome Mappings; genome sequencing; Genomics; Goals; Haploidy; Haplotypes; Head circumference; Health; Human; Human Genome; Impaired cognition; Individual; insight; Investigation; Knockout Mice; Laboratories; Lead; Learning; Letters; Libraries; Life; Link; Macrocephaly; Mammals; Maps; Memory; Mental Retardation; Microcephaly; Mole the mammal; mouse model; Mus; novel; Oligonucleotides; Paper; Patients; Pilot Projects; Rattus; Recurrence; Reporting; Role; Sampling; Schizophrenia; Syndrome; Tertiary Protein Structure; Testing; Universities; Washington

Project start date: 2009-07-02

Project end date: 2014-02-28

Budget start date: 1-MAY-2011

Budget end date: 29-FEB-2012

PFA/PA: PA-07-070

5R01MH081203-03 (2011): $428197

VITAMIN C TRANSPORTERS IN THE BRAIN

M James, Professor
Vanderbilt Universitycity: Nashville    country: United States (us)

Grant 5R01NS057674-04 from National Institute Of Neurological Disorders And Stroke

Abstract: Vitamin C, or ascorbic acid, has been shown in animal and clinical studies to help delay or prevent complications attributed to increased oxidant stress in stroke and in several neurodegenerative disorders. Normally, ascorbate is maintained at relatively high concentrations in brain and especially in neurons, where it serves as an antioxidant and supports several important neuronal functions. Ascorbate concentrations in the low millimolar range are generated in neurons by a specific ascorbate transporter, termed the SVCT2. Mice engineered to lack this transporter have very low brain ascorbate contents and die at birth with cerebral hemorrhage, indicating that the vitamin is crucial for survival. The overall goal of this proposal is to determine how the SVCT2 transporter is regulated in neurons, and whether changes in transporter number can affect neuronal function and antioxidant defenses, both in vitro and in vivo. There are three specific aims. In the first aim, mechanisms of SVCT2 regulation and structure-function relationships will be evaluated in cultured SH-SY5Y neuroblastoma cells and in Xenopus oocytes. Regulation of both SVCT2 expression and activity in response to oxidant stress will be assessed in SH-SY5Y cells. Further, the functional role of key cysteines on the extracellular surface of the protein will be evaluated by affinity labeling in SH- SY5Y cells and by mutagenesis approaches in Xenopus oocytes. In the second aim, the mechanism by which knock-out mice lacking the SVCT2 die at birth will be determined. Also in Aim 2, a novel model of oxidant stress due to combined deficiencies of vitamins E and C will be used to establish whether transgenic mice expressing increased amounts of SVCT2 are protected from neurologic damage compared to mice expressing normal or decreased (50%) SVCT2. In the third aim, primary culture neurons will be prepared from mice expressing several different levels of SVCT2 (transgenic, wild-type, heterozygous knock-out, and homozygous knock-out). These cells will be used to test whether SVCT2 expression 1) determines the ability of neurons to maintain intracellular ascorbate, 2) increases resistance of neurons to oxidant stress, 3) enhances maturation of cortical neurons in culture, and 3) affects neuronal electrophysiology. Results from these studies are expected to show that the SVCT2 ascorbate transporter, as regulated by oxidant stress, is crucial for neuronal function, development, and survival. This project to study the function, regulation and tissue expression of the transporter for vitamin C in the brain has relevance for neuronal function, maturation and resistance to oxidant stress. Several neurodegenerative diseases including Alzheimer´s and Parkinson´s are associated with increased oxidant stress so that this project will help define the role of vitamin C in oxidant injury to the brain

Keywords: Affect; affinity labeling; Affinity Labels; Alzheimer`s Disease; amidation; Animals; Antioxidants; ascorbate; Ascorbic Acid; Birth; Brain; Brain Death; Cells; Cerebral hemisphere hemorrhage; Clinical Research; Cysteine; Development; Electrophysiology (science); Engineering; extracellular; Glutamates; Goals; Health; Homeostasis; human disease; Huntington Disease; In Vitro; in vivo; Infarction; Injury; Knock-out; Knockout Mice; Membrane Proteins; Modeling; Mus; Mutagenesis; Myelin; Nervous System Physiology; Neuraxis; neuroblastoma cell; Neurodegenerative Disorders; Neurologic; Neurons; novel; oxidant stress; Oxidants; Oxidation-Reduction; Parkinson Disease; Peptides; prevent; Proteins; Regulation; Reperfusion Injury; Resistance; response; Role; sodium-dependent vitamin C transporter 2; stroke; Structure-Activity Relationship; Synapses; Testing; Tissues; Toxic effect; Transgenic Mice; Transgenic Organisms; Vitamin C in Brain Pathway; Vitamin E Deficiency; Vitamins; Xenopus oocyte

Project start date: 2008-07-01

Project end date: 2012-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PA-07-070

5R01NS057674-04 (2011): $296160

LESSONS FROM THE OR: USING CLINICAL BIOLOGIC CORRELATES TO INFORM HSV TRIAL

M James, Professor & Director Of Neurosurgery
University Of Alabama At Birminghamcity: Birmingham    country: United States (us)

Abstract: Using Clinical Biologic Correlates To Inform HSV Trial Design Through this Program Project grant we successfully generated and characterized several new, promising genetically engineered herpes simplex viruses (HSV) that are candidates for evaluation in clinical trials of patients with malignant glioma. These viruses were extensively studied and characterized in both in vitro and in vivo models to demonstrate proof-of-principle of safety and efficacy. Increased efficacy was provided by (i) expression of certain foreign genes and (ii) adjunctive therapy with radiation. This project will exploit our clinical trial experience in Phase I studies of G207, a first generation engineered oncolytic herpes simplex virus (oHSV) in the treatment of gliomas. We have developed extensive experience in the first use of a genetically engineered HSV for the treatment of human glioma in three Phase I clinical trials, with an upcoming study of a human IL-12 expressing engineered HSV-1 (M032) to begin imminently. We have collected a variety of tissue specimens to delineate the biologic response of human glioma patients treated with G207. We propose to use these human specimens for the following AIM 1 will determine if IL-12 expression by oncolytic HSV produces differences in the glioma tumor microenvironment and viral replication that impact tumor response. Evaluation of data from our G207 trials and upcoming M032 trial will be undertaken; response to therapy and corresponding findings will be evaluated for potential correlations. AIM 2 will determine whether HSV-treated tumors have SR, NSR or nu87 gene expression profiles defined by Project 2 and whether any of these GEPs are predictive of tumor response to treatment with ILI2 expressing OHSV, as well as identifying additional GEPs that are associated with poor outcome after treatment with IL- 12-expressing oHSV. Preclinical models emulating these responses will be developed to confirm the predictability of response to different viral constructs and adjunctive therapies. AIM 3 will determine the impact of glioma progenitor cells (GPC) on response to oncolytic HSV therapy by contrasting the fraction of GPC present with the response to virus therapy. Similarly we will assess the effects of IL-12 expression on GPC response. Aim 4 will serve as a planning aim for the next oncolytic HSV (after M032) to enter clinical trials that emerges from this Program Project. The aim includes decision analysis selection of the virus to be used, along with any adjuvant therapies; initiation of pretrial animal studies; and development of a clinical protocol, production of cGMP virus, as well as application for additional funding to actually perform the trial. RELEVANCE (See instructions) We have undertaken 3 Phase I clinical trials with a first generation oncolytic HSV in patients with malignant glioma and our results have confirmed our observations that tumor gene expression profile (GEP) defines susceptibility or resistance. Studies proposed here will analyze tumor tissues from these patients and from those to be treated with our second generation oHSV, expressing ILI 2. From these biologic studies, correlation of tumor GEP with response to oHSV will inform design of new oHSV that can overcome glioma resistance factors and broaden applicability of oHSV for efficacious anti-glioma therapy

Keywords: Adjuvant Therapy; Aftercare; Algorithms; Animals; Archives; base; Biology; cGMP production; Clinical; Clinical Protocols; Clinical Trials; Clinical Trials Design; Correlation Studies; Data; Decision Analysis; design; Development; Engineering; Evaluation; Exclusion Criteria; experience; Funding; G207; Gene Expression; Gene Expression Profile; Generations; Genes; Glioma; Herpesvirus 1, Human; Human; Immune response; improved; In Situ; In Vitro; in vivo Model; inclusion criteria; Instruction; Interleukin-12; Malignant Glioma; Molecular Profiling; novel; Oncolytic; Outcome; Patients; phase 1 study; Phase I Clinical Trials; Pre-Clinical Model; Predisposition; Program Research Project Grants; programs; R Factors; Radiation; Recurrence; Resistance; response; Role; Safety; Simplexvirus; Specimen; Stem cells; Tissues; treatment planning; tumor; Tumor Tissue; Validation; Viral; Virus; Virus Replication

Relevance: We have undertaken 3 Phase I clinical trials with a first generation oncolytic HSV in patients with malignant glioma and our results have confirmed our observations that tumor gene expression profile (GEP) defines susceptibility or resistance. Studies proposed here will analyze tumor tissues from these patients and from those to be treated with our second generation oHSV, expressing IL12. From these biologic studies, correlation of tumor GEP with response to oHSV will inform design of new oHSV that can overcome glioma resistance factors and broaden applicability of oHSV for efficacious anti-glioma therapy

Project start date: 2011-07-01

Project end date: 2014-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

5P01CA071933-13_5095 (2011): $175290

A CELL PHONE-BASED STREET INTERSECTION ANALYZER FOR VISUALLY IMPARED PEDESTRIANS

M James, Associate Scientist
Smith-kettlewell Eye Research Institutecity: San Francisco    country: United States (us)

Grant 5R01EY018345-03 from National Eye Institute

Abstract: Urban intersections are the most dangerous parts of a blind person´s travel. They are becoming increasingly complex, making safe crossing using conventional blind orientation and mobility techniques ever more difficult. To alleviate this problem, we propose to develop and evaluate a cell phone-based system to analyze images of street intersections, taken by a blind or visually impaired person using a standard camera cell phone, to provide real-time feedback. Drawing on our recent work on computer vision algorithms that help a blind person find crosswalks and other important features in a street intersection, as well as our ongoing work on cell phone implementations of algorithms for indoor wayfinding and for reading digital appliance displays, we will refine these algorithms and implement them on a cell phone. The information extracted by the algorithms will be communicated to the user with a combination of synthesized speech, audio tones and/or tactile feedback (using the cell phone´s built-in vibrator). Human factors studies will help determine how to configure the system and its user controls for maximum effectiveness and ease of use, and provide an evaluation of the overall system. The street intersection analysis software will be made freely available for download into any camera-equipped cell phone that uses the widespread Symbian operating system (such as the popular Nokia cell phone series). The cell phone will not need any hardware modifications or add-ons to run this software. Ultimately a user will be able to download an entire suite of such algorithms for free onto the cell phone he or she is already likely to be carrying, without having to carry a separate piece of equipment for each function. Relevance The ability to walk safely and confidently along sidewalks and traverse crosswalks is taken for granted every day by the sighted, but approximately 10 million Americans with significant vision impairments and a million who are legally blind face severe difficulties in this task. The proposed research would result in a highly accessible system (with zero or minimal cost to users) to augment existing wayfinding techniques, which could dramatically improve independent travel for blind and visually impaired persons

Keywords: Accidents; Address; Algorithms; American; assistive device; Assistive Technology; base; biological signal transduction; blind; blind individual; blind people; blind person; Cell Communication and Signaling; Cell Phone; Cell Signaling; Cellular Phone; Complex; computer program/software; Computer Programs; Computer software; computer vision; Computer Vision Systems; consumer product; cosmetic product; Cosmetics; cost; Custom; Detection; Devices; digital; Educational Mainstreaming; Effectiveness; Equipment; Evaluation; experience; Face; facial; Feedback; Figs; Figs - dietary; Grant; Human; Human, General; Image; Image Analyses; Image Analysis; image evaluation; image processing; imaging; Impairment; improved; Infrastructure; Intracellular Communication and Signaling; Left; legally blind; Light; Mainstream Education, achievement; Mainstreaming; Mainstreaming (Education); Man (Taxonomy); Man, Modern; Marketing; Methods and Techniques; Methods, Other; Modification; novel; Operating System; Partial Sight; Pattern; Performance; Photoradiation; prevent; preventing; programs; Programs (PT); Programs [Publication Type]; Reading; Research; Research Infrastructure; Resolution; ROC Analysis; Running; Self-Help Devices; Series; Sight; Signal Transduction; Signal Transduction Systems; Signaling; skills; Software; spatial navigation; Speech; System; System, LOINC Axis 4; Tactile; Target Populations; Techniques; Telephone, Cellular; Testing; Time; tool; trafficking; Training; Travel; Vision; Vision Systems, Computer; Vision, Diminished; Vision, Low; Vision, Reduced; Vision, Subnormal; Visual; Visual impairment; visually impaired; visually impaired people; Visually Impaired Persons; Walking; way finding; wayfinding; web site; Work; Zebra

Project start date: 2007-09-01

Project end date: 2011-02-28

Budget start date: 1-SEP-2009

Budget end date: 28-FEB-2011

5R01EY018345-03 (2009): $346662

BIOCHEMICAL ANALYSES OF TYPE II DNA TOPOISOMERASES

M James, Professor
University Of California Berkeleycity: Berkeley    country: United States (us)

Grant 5R01CA077373-13 from National Cancer Institute

Abstract: Type II topoisomerases are a ubiquitous class of proteins that use ATP to actively transport one DNA duplex through another. This reaction is essential for supercoiling homeostasis and resolving cytotoxic chromosome tangles prior to cell division. Type II topoisomerases are also targeted by drugs that serve as frontline clinical therapies for cancer and bacterial infections. The long-term objective of this proposal is to investigate the molecular basis of type II topoisomerase function and drug inhibition. Although rough framework for the catalytic cycle of these enzymes is in place, there remain many critical questions surrounding the mechanisms by which type II topoisomerases discriminate between different topological states of DNA, undergo allosteric transitions to drive DNA transport, and are inhibited by small molecule "poisons" that stimulate DNA cleavage. Using a combination of structural, biochemical, and biophysical methods we aim to fill these gaps by 1) Determining the structure of a "poisoned" type II topoisomerase/DNA complex, 2) Establishing how a novel DNA binding and bending domain in bacterial type II topoisomerases controls substrate selectivity and functional output, and 3) Defining the molecular mechanisms and kinetics of DNA deformations and key structural rearrangements in the topoisomerase catalytic cycle. Our proposed studies will define the physical events by which type II topoisomerases facilitate the passage of one DNA segment through another to globally control DNA topology, and by which anticancer and antibacterial inhibitors subvert enzyme function. Data resulting from such efforts broadly impact a number of important scientific fronts, from understanding the dynamics of ATP-dependent molecular machines and regulation of chromosome superstructure, to defining the physical action of anti-topoisomerase therapies and aiding drug development. Type II topoisomerases are molecular machines that disentangle DNA, as well as validated targets for frontline antibacterial and anticancer therapies. This proposal aims to understand the biochemistry and mechanics of the topoisomerase reaction, and to determine how some of the most widely-used anti-topoisomerase drugs block enzyme function

Keywords: Address; Affinity; Anti-Bacterial Agents; Architecture; Bacteria; Bacterial Infections; Bacterial Typing; base; Binding (Molecular Function); Biochemical; Biochemistry; Biology; C-terminal; Calculi; cancer therapy; Cell Death; Cell division; cell killing; Cell physiology; Cell Survival; Chemicals; Chromosomes; Cleaved cell; Clinical; Communicable Diseases; Complex; Coupled; cytotoxic; Data; dimer; Discrimination (Psychology); DNA; DNA Binding; DNA biosynthesis; DNA Double Strand Break; DNA Sequence Rearrangement; DNA Topoisomerase IV; Drug Delivery Systems; drug development; Enzymes; Event; Exhibits; Genetic Transcription; Homeostasis; Image; improved; in vivo; inhibitor/antagonist; innovation; inorganic phosphate; insight; interest; Kinetics; Malignant Neoplasms; Mechanics; Methods; Microfluidics; Molecular; Molecular Machines; Mutagenesis; Neurofibrillary Tangles; novel; Output; paralogous gene; Pharmaceutical Preparations; Poisoning; Poisons; Problem Solving; Protein Isoforms; Proteins; Reaction; Regulation; Research; segregation; small molecule; Structure; Structure-Activity Relationship; Superhelical DNA; Testing; Time; Topoisomerase; Topoisomerase II; Topoisomerase Inhibitors; Treatment Efficacy; Work

Project start date: 1999-05-01

Project end date: 2014-02-28

Budget start date: 1-MAR-2011

Budget end date: 29-FEB-2012

PFA/PA: PA-07-070

5R01CA077373-13 (2011): $260468

ACTION OF THE SV40 T ANTIGEN CHAPERONE MACHINE ON TUMOR SUPPRESSORS

M James, Professor
University Of Pittsburgh At Pittsburghcity: Pittsburgh    country: United States (us)

Grant 5R01CA120997-06 from National Cancer Institute

Abstract: Tumor suppressors are regulatory proteins that receive and integrate diverse signals and function to exert control over key cellular processes such as cell proliferation, differentiation, and apoptosis. Because loss or perturbation of their activity often results in cancer or other diseases, and because of their central role in governing organismal development and tissue homeostasis, these proteins are of great interest. The retinoblastoma protein (pRb) is a well characterized tumor suppressor that, in concert with two related proteins, p130and p107,control cell cycle entry and exit, in part, by regulating the activity of the E2F family of transcription factors. Many viruses, including Simian virus 40 (SV40) encode oncoproteins that bind Rb-family members and interfere with their ability to regulate E2Fs. The large tumor antigen (T antigen) encoded by SV40 binds to pRb, p107, and p130via an LXCXE motif and blocks the ability of these proteins to inhibit E2F-dependent transcription and to induce growth arrest. The retinoblastoma family has been studied intensively, yet little is known about the molecular basis by which viruses, such as SV40, block their action. In fact, interaction with T antigen has different consequences for each Rb protein. For example, p130 is degraded following SV40 infection or transformation, while the levels of pRb remain unchanged. Thus, T antigen appears to be able to distinguish different Rb-E2F complexes, but the basis for this discrimination is unknown. Like many regulatory proteins pRb and E2F transcription factors do not exist in isolation. Rather they function as part of large multiprotein assemblages that include chromatin modifiers, the basal transcription apparatus, as well as other factors, and the dynamic assembly and disassembly of these complexes is critical to their regulation. T antigen has a J domain and has been shown to function as a DnaJ molecular chaperone. The J domain is required for a vital DNA replication, transcriptional control, and virion assembly. Importantly, the J domain is required for T antigen to block the function of Rb proteins and thus to activate E2F-dependent transcription. This application seeks to understand the mechanistic and structural basis for the action of T antigen´s recognition and disruption of Rb-E2F complexes.´ First, biochemical studies will explore the ability of the T antigen chaperone machine to distinguish and act upon p130-E2F4-DP1, pRb- E2F4-DP1, and pRb-E2F1-DP1 complexes. Second, the role of J domain orientation and flexibility will be examined using a combination of NMR and X-ray crystallography. Finally, a combined genetic and biochemical approach will be used to identify additional protein participants in the chaperone reaction. These studies will enhance our understanding of how these tumor suppressors govern cell proliferation and survival, and how subversion of these mechanisms by viruses or genetic mutation, contribute to cancer

Keywords: Adopted; Amino Acids; antigen binding; Apoptosis; ATP Hydrolysis; ATP phosphohydrolase; base; Binding (Molecular Function); Biochemical; Biological; Cell Cycle Regulation; Cell physiology; Cell Proliferation; Cell Survival; Chemicals; Chromatin; Complex; Crystallization; Crystallography; Data; Development; Discrimination (Psychology); Disease; DNA biosynthesis; E2F transcription factors; E2F1 gene; Family; Family member; flexibility; Gene Mutation; Genes; Genetic; genetic regulatory protein; Genetic Transcription; Goals; Growth; Homeostasis; Homologous Gene; Host Defense; In Vitro; in vivo; Infection; interest; Location; Malignant Neoplasms; Mediating; member; metaplastic cell transformation; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; mutant; Oncogene Proteins; Participant; Phosphorylation; Play; Production; Promotor (Genetics); Protein Binding; Proteins; Reaction; Recruitment Activity; Regulation; Relaxation; research study; Resolution; Rest; Retinoblastoma; Retinoblastoma Protein; Role; sialosyl-T antigen; Signal Transduction; Simian virus 40; Staging; System; Testing; three dimensional structure; Tissues; transcription factor; Transcriptional Activation; Transcriptional Regulation; Tumor Antigens; Tumor Suppressor Proteins; Viral; viral DNA; Viral Tumor Antigens; Virion; Virus; Virus Diseases; Work; X-Ray Crystallography

Project start date: 2006-04-14

Project end date: 2012-02-29

Budget start date: 1-MAR-2011

Budget end date: 29-FEB-2012

5R01CA120997-06 (2011): $322373

RESEARCH TRAINING IN DIABETES AND ENDOCRINOLGY

M James, Professor
Vanderbilt Universitycity: Nashville    country: United States (us)

Grant 2T32DK007061-37 from National Institute Of Diabetes And Digestive And Kidney Diseases

Abstract: This proposal seeks continued support for an Institutional NRSA that provides post- doctoral research training to 4 M.D. and/or Ph.D. trainees in endocrinology, diabetes, and metabolism. Also requested are 6 short-term summer medical student training positions. This multi-disciplinary program involves 27 faculty preceptors in six clinical and basic science departments of Vanderbilt University School of Medicine. It is an integral component of the training activities of the Vanderbilt Diabetes Research and Training Center. Its goal is to provide trainees with the knowledge and skills required for independent clinical or basic science research careers. M.D. trainees are encouraged to acquire a firm foundation in the basic sciences, but also to select projects in translational or clinical research areas. Ph.D. trainees are urged to relate their research to health- oriented problems. Trainees are selected from the nationwide pool of applicants that traditionally seek training at Vanderbilt. M.D. trainees usually have three to four years of residency training before entering the program. Ph.D. trainees have typically just received or are one year out from their doctorate. Trainees are selected on the basis of pre-doctoral performance and especially on their potential as future researchers and educators. Training positions are allocated approximately equally between M.D.´s and Ph.D.´s. The training program utilizes the preceptor approach, in which the trainee develops a research project under the guidance of a faculty preceptor. Several criteria are used to match the preceptor with the trainee, including the expressed interests of the trainee, quality of the project proposed, amount of direct supervision that the preceptor can provide, and adequacy of the research facilities available. Typical research topics include hormone action in humans as it relates to diabetes, intermediary metabolism, micronutrient effects on metabolic disease, metabolic regulation, molecular genetics of metabolic diseases, and translation of diabetes care delivery. Research training is supplemented by conferences, seminars, and required and optional coursework. Trainee progress is evaluated by a mentoring committee twice-yearly and the program is evaluated yearly by an oversight committee. In the 36 years of NIH support, the program has supported 108 trainees. Over 80% of those who have completed training have chosen careers in academia, industry, or government. We are training the next generation of scientists with expertise in diabetes, metabolic, and endocrine diseases. These trainees will make discoveries during and after their training that will improve our understanding of and lead to new and improved treatment for these conditions

Keywords: Diabetes Mellitus; Research Training

Relevance: We are training the next generation of scientists with expertise in diabetes, metabolic, and endocrine diseases. These trainees will make discoveries during and after their training that will improve our understanding of and lead to new and improved treatment for these conditions

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

2T32DK007061-37 (2011): $273500

3T32DK007061-37S1 (2011): $74365

INFECTION BIOLOGY TRAINING GRANT

M James, Associate Professor
University Of Illinois Urbana-champaigncity: Champaign    country: United States (us)

Grant 5T32AI078876-02 from National Institute Of Allergy And Infectious Diseases

Abstract: This proposal is a request for funding to establish an institutional pre-doctoral training program in Infection Biology at the University of Illinois at Urbana-Champaign submitted by an interdisciplinary program faculty from the Departments of Microbiology and Pathobiology. The expertise of the program faculty in microbial physiology, genetics, pathogenesis, immunology, and ecology of infectious diseases will provide broad graduate training in modern molecular pathogenesis that goes beyond what is available in current graduate programs. The 19 productive researchers that make up the training faculty are highly interactive with established collaborations that cross departmental and college boundaries. The broad interdisciplinary training of these students will enhance the research efforts funded by grants from the NIH and other agencies. The training program will also facilitate interactions with scientists at other institutions by sponsoring a seminar series, by participating in a locally sponsored international conference on New and Re- emerging Infectious Diseases, and by providing support for students to attend additional national meetings to present their data to a critical audience. We request funds to support four trainees. These pre-doctoral students will be supported for a two-year period starting at the end of their second year of graduate studies, after they have completed much of their coursework, have established a thesis project, and have generated significant preliminary data. The program will enhance the training and mentorship of these students. RELEVANCE (See instructions) Infectious diseases remain the third leading cause of death in the United States and the second leading cause of death worldwide. Funding of this training program will allow us to continue to build on our tradition of training high-quality scientists and to foster a growing community of interactive biologists working on high-impact timely problems across all areas of infectious disease

Keywords: Biology; Grant; Infection; Training

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-08-226

5T32AI078876-02 (2011): $177164

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IMMUNE POTENTIAL OF ANIMAL LACKING CLASS IA MOLECULES

M James
University Of Texas Sw Med Ctr/dallascity: Dallas    country: United States (us)

Grant 5R01AI045764-10 from National Institute Of Allergy And Infectious Diseases

Abstract: This proposal investigates a unique function for CDS T cells in the immune response against infectious agents. Our major focus is on the role that CDST cells play in the innate immune response during infection with Listeria monocytogenes (LM). We recently demonstrated that antigen non specific CDS T memory cells participate in a cytokine driven innate response against LM by secreting interferon-y. Further ransfer of "innate" CDS T cells into interferon-y deficient mice protects them from infection with LM. We propose that CDST cells play a major role in the INF-y mediated innate response. In the first Aim we investigate the relative potency of CDS vs NK cells in providing this type of innate protection. We postulate that effector CDS T memory cells (TFM). which have been shown to play a minor role in the adaptive immune response play an important role in the innate response and will test thii hypothesis. In the second Aim we will examine the localization of CDS central memory cells (TOM). TEM. anc MK cells at sites of LM infection in spleen and liver. We will use mice deficient in CCR7 binding chemokine! CCL-19 and -21) to assess how this chemokine-receptor interaction affects CDS T cells in the innate response. We will also utilize new BAG transgenic mice that express Thy-1.1 as a reporter for IFN-y secretion to examine IFN-y secreting CDS and NK cells in situ. In the third Aim we will determine the role of CDS T cells in polarizing nai´ve CD4 T cells to the Th1 subset. In the fourth Aim we will examine by microarray analysis the gene display of CDS T cells that are activated by IL-12/18 (innate) vs those that are activated through the TcR (adaptive). In summary, this proposal will add important new information on how CDS T cells function in the innate immune response against intracellular pathogens

Keywords: Affect; Animals; Antigens; Area; ATGN; Binding; Binding (Molecular Function); body system, hepatic; CD4 lymphocyte; CD4 Positive T Lymphocytes; CD4 T cells; CD4+ T cell; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; Cell Function; Cell physiology; Cell Process; Cells; Cells, CD4; Cellular Function; Cellular Physiology; Cellular Process; chemoattractant cytokine; chemokine; chemokine receptor; cytokine; Cytokines, Chemotactic; Cytotoxic cell; cytotoxicity; Data; Genes; helper T cell; Homologous Chemotactic Cytokines; host response; IFN; Immune; Immune response; Immunity; Immunity, Innate; Immunity, Native; Immunity, Natural; Immunity, Non-Specific; immunogen; immunoresponse; In Situ; Infection; Infectious Agent; infectious organism; Intercrines; Interferons; K lymphocyte; L. monocytogenes; Listeria monocytogenes; Liver; Lymphatic Tissue; Lymphoid Tissue; Lytotoxicity; Mammals, Mice; Mediating; Memory; memory T lymphocyte; Mice; Microarray Analysis; microarray technology; Microarray-Based Analysis; Minor; Molecular Interaction; Monitor; Murine; Mus; Natural Immunity; Natural Killer Cells; NK Cells; Organ; organ system, hepatic; pathogen; pathway; Pathway interactions; Phenotype; Play; Population; Red Pulp; Relative; Relative (related person); Reporter; response; Reticuloendothelial System, Spleen; Role; SIS cytokines; Site; social role; Spleen; Splenic Red Pulp; Subcellular Process; T memory cell; T-Cells; T-Lymphocyte; T4 Cells; T4 Lymphocytes; TEM cells; terminally differentiated effector memory (TEM) T cells; terminally differentiated effector memory T cells; Testing; thymus derived lymphocyte; Thymus-Dependent Lymphocytes; trafficking; Transgenic Mice

Project start date: 1999-09-01

Project end date: 2011-12-31

Budget start date: 1-JAN-2010

Budget end date: 31-DEC-2011

PFA/PA: PA-04-119

5R01AI045764-10 (2010): $330250

DEVELOPMENT OF TARGETED ANTICANCER DRUGS

M James, Professor
Mount Sinai School Of Medicinecity: New York    country: United States (us)

Grant 5R01CA127963-05 from National Cancer Institute

Abstract: This project is concerned with the development of a new drug development paradigm that focuses on the use of preclinical studies in animals to derive predictive pharmacokinetic [PK] - pharmacodynamic [PD] models that can aid in the rational selection of drug candidates, and be used to predict human PK-PD characteristics. A unique feature of the proposal is that the drug´s PK-PD properties in target tissues, in this case brain and brain tumors, are integral to the drug development plan. The proposed drug development strategy will be demonstrated for a series of epidermal growth factor receptor [EGFR] inhibitors, a class of small molecular weight compounds that are represenative of targeted anticancer drugs, an emerging anticancer therapeutic strategy. The motivation for this new drug development approach is based on limitations of current semi-empirical anticancer drug development approaches, which rely on a plethora of drug efficacy studies that lack predictive capability on how drugs should be used in humans. Moreover, EGFR inhibitors are representative of the problems of defining optimal doses for targeted therapies in cancer patients, since these classes of drugs often do not possess readily identified dose-limiting toxicites that typically guide the use of standard cytotoxics. The PK-PD driven approach will rely on the use of hybrid PK-PD models that enables a target tissue [i.e. brain tumors] to be represented in physiological terms, which accomodates a mechanistic depiction of drug disposition, and facilitates model scale-up to humans. The drug development scheme will be implemented in brain tumor models that differ only in EGFR status, one being wild-type and the other the vIII mutant, which is associated with drug sensitivity, which will allow us to test the broader applicability of the approach to other brain tumor models whose molecular characteristics are known. A synopsis of the Specific Aims are 1) screen new EGFR inhibitors for brain accumulation in a cassette dosing format; 2) determine in vitro PD endpoints associated with EGFR signaling pathways; 3) develop hybrid PK-PD models in mice; and 4) evaluate drug efficacy in a range of brain tumor types based on the use of PK-PD equivalent dosing regimens. The proposed drug development package relies on a drug´s in vivo pharmacological behavior to screen, select, and subsequently design therapeutic regimens based on model-predicted PK-PD endpoints. It is believed that this approach offers the best opportunity to develop drugs in a rational and optimal manner. The translation of preclinical pharmacological information of new anticancer drugs to patients does not offer a direct conduit to ensure biologically relevant and optimal drug doses are administered. The current proposal will attempt to rectify these deficiencies through the implementation of a new drug development paradigm based on a drug´s pharmacokinetic and pharmacodynamic characteristics in the target tissue, which for this project is brain tumors

Keywords: Animals; anti-cancer therapeutic; Antineoplastic Agents; Area Under Curve; base; Behavior; Biological Assay; Blood - brain barrier anatomy; Brain; Brain Neoplasms; Bypass; Cancer Patient; Cell Line; Cells; Characteristics; Clinical; Clinical Trials; Coupled; cytotoxic; cytotoxicity; Data Set; design; Development; Development Plans; Dose; Dose-Limiting; Drug Administration Schedule; drug candidate; drug development; drug discovery; drug efficacy; Drug Efflux; Drug Kinetics; Drug or chemical Tissue Distribution; drug sensitivity; efflux pump; Ensure; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; epidermal growth factor receptor VIII; Erlotinib; Failure (biologic function); Gefitinib; Glioma; Goals; Health; Human; Hybrids; In Vitro; in vitro activity; in vivo; Individual; Inhibitory Concentration 50; Investigation; Lead; Letters; Link; Measurement; Measures; Modeling; Molecular; Molecular Weight; Motivation; Mus; mutant; novel; Oral; Pathway interactions; Patients; Penetration; Pharmaceutical Preparations; pharmacodynamic model; Pharmacodynamics; Pharmacogenetics; Phase; Physiological; Plasma; pre-clinical; preclinical study; Procedures; programs; Property; protein expression; Protocols documentation; Receptor Signaling; Regimen; Reliance; Route; scale up; Schedule; Scheme; Screening procedure; Series; Signal Pathway; Site; Testing; Therapeutic; therapeutic target; Time; Tissues; tool; Translations; tumor; Tumor Cell Line; Tumor-Derived; Tyrosine Kinase Inhibitor; uptake

Project start date: 2008-07-01

Project end date: 2013-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-07-070

5R01CA127963-05 (2011): $316542

MOLECULAR THERAPY FOR CF AND GENETIC DISEASES

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Grant 5P30DK047757-18 from National Institute Of Diabetes And Digestive And Kidney Diseases

Abstract: This is a competitive renewal for Penn´s P30 NIDDK Center on Molecular Therapy for Cystic Fibrosis and Genetic Diseases which has been in place since 1993. At the last renewal we shifted our strategy from a focus on clinical trials to vector discovery and distribution with an increased emphasis on immunology and animal models. This has had the desired result of substantially fueling a robust pipeline of promising preclinical programs that are entering clinical trials with better technology and an enhanced understanding of vector biology necessary for clinical success. With encouragement from NIDDK, we expanded access to our Cores to investigators outside of Penn to establish important strategic collaborations. During this cycle of the grant, Vector Core services increased from 576 in year 11 of the grant to 1458 projected for the current year (year 15). The substantial expansion of gene and cell therapy research associated with our Center is due in large part to the availability of new AAV and lentiviral technology discovered at Penn and distributed through our Vector Core. An important aspect of this expansion has been in the area of immunology that has emerged as a major impediment to success. The Center has aggressively recruited the participation of scientists interested in applied immunology and genetic vaccines to focus on issues relevant to gene and cell therapy. The current research base is substantial and strong across all areas relevant to the Center. Current total annual direct costs of grants awarded to Center Participants has been organized into the following three areas Genetic Diseases, Stem Cells and Gene Transfer - $49 mil; Cystic Fibrosis - $8.6 mil; and Immunology and Vaccines -$14.9 mil. The Pjlot Grant program remains strong and has been useful in supporting young investigators and encouraging the participation of new investigators. Important recruitments at Penn have provided the opportunity to enhance leadership of the Center´s Cores. This application requests support for Pilot Grants as well as the following Cores Vector, Immunology, Cell Morphology and Animal Models

Project start date: 1997-09-30

Project end date: 2014-03-31

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

PFA/PA: RFA-DK-07-010

5P30DK047757-18 (2011): $1058170

HUMAN CYTOMEGALOVIRUS NUCLEAR EGRESS: MOLECULAR MECHANISMS AND DRUG TARGETING

M James, Pd/pi
Harvard University (medical School)city: Boston    country: United States (us)

Grant 5R01AI026077-24 from National Institute Of Allergy And Infectious Diseases

Abstract: This application´s long-term objectives are to understand the central steps by which human cytomegalovirus (HCMV) nucleocapsids transit from the nucleus to the cytoplasm (nuclear egress), and to exploit that understanding for drug discovery. This project is especially health-related, as new drugs are needed for treatment of HCMV. In this application, HCMV proteins that are involved in nuclear egress are investigated. One of these proteins, the UL97 protein kinase, is already an established drug target. Two other proteins, UL50 and UL53, interact to form a nuclear egress complex (NEC) that can serve as a new drug target. Specific aim 1 is to investigate the roles of UL97 that are important for production of infectious virus in both non-dividing and dividing cells; in particular, whether the only role of UL97 in nuclear egress is phosphorylation of lamin A/C to disrupt the nuclear lamina. A principal approach will be to analyze a recombinant HCMV expressing a dominant negative mutant of lamin A/C in place of UL97 to determine if that virus can replicate and disrupt nuclear lamina as well as wild type virus in dividing cells or as well as a virus expressing human papillomavirus E7 in non-dividing cells. Specific aim 2 is to investigate the function(s) of the NEC. HCMV mutants that fail to express UL50 or UL53 or that have more specific alterations such as defects in UL50-UL53 interactions will be constructed and their block(s) in nuclear egress will be determined with the aid of techniques including electron microscopy. Why the HCMV NEC is not sufficient to disrupt nuclear lamina in the absence of UL97 will be investigated. Candidate proteins that interact with the NEC in HCMV-infected cells will be investigated to explore the hypothesis that such proteins are recruited to effect budding of nucleocapsids through the inner nuclear membrane. These proteins will be tested for co-localization with the NEC in cells, whether they interact directly with the NEC, and, if so, to map determinants of the interaction. The importance of these proteins for HCMV replication will be investigated using techniques including RNA interference. Should these studies fail to uncover a role for interacting proteins, in vitro studies to test whether the NEC can promote membrane curvature and vesiculation will be undertaken. Specific aim 3 is to determine the structure of the NEC. The structures of truncated versions of NEC subunits that retain sequences that are conserved among herpesviruses are being determined by NMR, as are the locations of the subunit interfaces, which can lead to an NMR structure of the complex. Efforts to improve crystals of the complex will continue, with the goal of obtaining a high resolution crystal structure. Specific aim 4 is to establish a high throughput assay for small molecules that inhibit subunit interactions of the NEC using an amplified luminescent proximity homogeneous assay or other assay. This assay will be used to screen libraries of small molecules and natural products. "Hits" will be then assayed for biochemical activity and specificity, and for anti-HCMV activity and cytotoxicity, with the long range goal of developing them into antiviral drugs. HCMV causes severe disease in people with impaired immunity, and is associated with a number of diseases in the immunocompetent population. There is considerable need for new drugs to combat HCMV, as current drugs have major limitations. The research proposed should not only provide information that could aid in understanding drug targets and mechanisms, but aims directly to discover new anti-HCMV drugs

Keywords: Antiviral Agents; Applications Grants; base; Biochemical; Biochemistry; Biological; Biological Assay; Biological Factors; Cell Nucleus; Cell physiology; Cells; combat; Complex; Conserved Sequence; Cyclin-Dependent Kinases; Cytomegalovirus; Cytoplasm; cytotoxicity; Defect; design; Disease; Dominant-Negative Mutation; Drug Delivery Systems; drug discovery; Drug effect disorder; drug mechanism; Drug resistance; Electron Microscopy; Enzymes; Exhibits; fascinate; Ganciclovir; Goals; Grant; Health; Herpesviridae; high throughput screening; Human; Human Papillomavirus; Immunity; Immunocompetent; improved; In Vitro; Infection; inhibitor/antagonist; insight; interest; Interphase Cell; Lamin Type A; Lead; Location; Maps; maribavir; Membrane; Membrane Proteins; Molecular; Molecular Genetics; Murid herpesvirus 1; mutant; Nuclear; Nuclear Inner Membrane; Nuclear Lamina; Nuclear Protein; Nuclear Structure; Nucleocapsid; pathogen; Pharmaceutical Preparations; Phosphorylation; Population; Process; Production; Protein Kinase; Proteins; public health relevance; Reagent; Recombinants; Recruitment Activity; Research; Resistance; Resolution; Retinoblastoma; Retinoblastoma Protein; RNA Interference; Role; Simplexvirus DNA polymerase; small molecule; small molecule libraries; Specificity; Structure; Techniques; Testing; tumor; Ursidae Family; Viral; Virus; Virus Replication; Work; X-Ray Crystallography

Relevance: Relevance HCMV causes severe disease in people with impaired immunity, and is associated with a number of diseases in the immunocompetent population. There is considerable need for new drugs to combat HCMV, as current drugs have major limitations. The research proposed should not only provide information that could aid in understanding drug targets and mechanisms, but aims directly to discover new anti-HCMV drugs

Project start date: 1988-04-01

Project end date: 2015-08-31

Budget start date: 1-SEP-2011

Budget end date: 31-AUG-2012

PFA/PA: PA-10-067

5R01AI026077-24 (2011): $620503

DNA VIRUS AS VECTORS FOR CARDIOVASCULAR DISEASES

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Grant 5P01HL059407-13 from National Heart, Lung, And Blood Institute

Abstract: This application requests support for the third cycle of this program project that has been dedicated to the development of gene therapy for cardiovascular diseases. We plan in this cycle to focus on the pre-clinical development of gene therapy to liver for the treatment of two severe dyslipidemias familial hypercholesterolemia (FH) and abetalipoproteinemia (ABL). In studies performed during the current cycle of this grant we developed a number of vectors based on novel adeno-associated viruses (AAV). We showed that many of these have substantially improved performance profiles in terms of gene transfer efficiency, safety and immunogenicity. The goal of this application is to identify the optimal AAV vector for liver directed gene therapy and to study areas of vector biology relevant for the potential testing of these vectors in humans. The feasibility of treating FH and ABL will be assessed in relevant animal models. Key milestones are identification of the clinical candidate vector in year one and the development of sufficient pre-clinical data at the end of year 5 to proceed with phase I clinical trials in both FH and/or ABL in a subsequent application. Project I, directed by Dr. J. Wilson, will develop the clinical candidate vector and will evaluate key issues of vector immunology relevant to safety and efficacy. Project II, directed by Dr. D. Rader, will perform all pre-clincal studies of gene therapy in animals models of FH (mice, rabbits and monkeys) and ABL (mice). Project III, directed by Dr. G. Gao, will evaluate the molecular biology of AAV in liver in terms of recombination with endogenous virus, integration and oncogenicity. The Projects will be supported by three scientific cores - Vector (Core A), Cell Morphology (Core B), and Animal Models (Core C) - and the Administrative Core (Core D). Lay . This project will use a novel viral vector system based on adeno-associated viral vectors (AAV) to develop gene therapy for two very severe inherited forms of lipid metabolism called familial hypercholesterolemia (FH) and abetalipoproteinemia (ABL). The grant will do all pre-clinical studies necessary to begin clinical trials

Project start date: 2000-05-15

Project end date: 2014-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

5P01HL059407-13 (2011): $2187963

CLINICAL AND COMPUTATIONAL STUDIES OF DOPAMINE FUNCTION IN SCHIZOPHRENIA

M James, Professor
University Of Maryland Baltimorecity: Baltimore    country: United States (us)

Grant 5R01MH080066-04 from National Institute Of Mental Health

Abstract: This 5 year proposal involving 3 universities explores the role of dopamine (DA) in the reinforcement learning working memory (WM), and decision making deficits of patients with schizophrenia (SZ) through behavioral experiments and computational modeling. While DA dysfunction is thought to be a fundamental aspect of SZ, rapid advances in the basic neuroscience understanding and modeling of the role of DA in reward processing remain to be translated into clinical research. The goal of this proposal is to provide an integrative account of how DA abnormalities may underlie the disabling cognitive/motivational deficits of SZ within a comprehensive computational framework that addresses the role of DA signaling in the basal ganglia and frontal cortex in action selection, learning, and WM. The experiments provide tests of model predictions of the types of impairments that result from the DA abnormalities that are thought to occur in SZ. In Aims 1-3, we test for deficits in processing of positive feedback signals that would be expected to result from abnormal increases of tonic DA across tasks tapping habit learning, decision making, and WM. We also test the novel, model based prediction that DA abnormalities in the orbital frontal cortex will result in impairment in processing the relative magnitudes of rewards and punishments with experiments designed where this will lead patients to perform at superior or inferior levels relative to controls. Aim 4 addresses the impact of antipsychotic treatment through the testing of patients both before and after the initiation of treatment where we will test the hypothesis that treatments facilitate learning from negative reinforcement as predicted by the model. These behavioral results will then be explored using computational modeling to determine the fit of observed behavior to predictions based on models where we have explored the role of increasing and decreasing different aspects of DA signaling to approximate the hypothesized abnormalities in SZ. Relevance Most SZ patients experience deficits in cognitive and motivational processing, resulting in significant disability. It is likely that abnormalities in the DA system may be involved in both types of impairments. Because all known antipsychotic treatments impact the DA system, the proposed work promises to increase understanding of how current treatments may have beneficial as well as possible adverse effects on these crucial areas of impairment

Keywords: 3, 4-Dihydroxyphenethylamine; 4-(2-Aminoethyl)-1, 2-benzenediol; Accounting; Address; Adverse effects; Antipsychotic Agents; Antipsychotic Drugs; Antipsychotics; Area; Basal Ganglia; Basal Nuclei; base; Behavior; Behavior Control; Behavioral; behavioral control; Behavioral Manipulation; Behavioral Paradigm; biological signal transduction; Cell Communication and Signaling; Cell Signaling; Chronic Schizophrenia; Clinical Research; Clinical Study; Cognition; Cognitive; computational framework; computational modeling; computational models; computational simulation; computational studies; computer based models; computer framework; Computer Simulation; computer studies; computerized modeling; Computerized Models; computerized simulation; Data; Decision Making; dementia praecox; design; designing; disability; Dopamine; dopamine system; drug/agent; Drugs; Dysfunction; experience; experiment; experimental research; experimental study; Exposure to; failure; Failure (biologic function); Feedback; FLR; frontal cortex; frontal lobe; Functional disorder; Goals; habit learning; heavy metal lead; heavy metal Pb; Hydroxytyramine; Impairment; in silico; Inferior; Intracellular Communication and Signaling; language translation; Lateral; Lead; Learning; Lesion; Major Tranquilizers; Mathematical Model Simulation; Mathematical Models and Simulations; Medial; Mediating; Medication; Memory Deficit; Memory impairment; Memory, Immediate; Memory, Short-Term; Memory, Shortterm; Modeling; Models, Computer; Motivation; motivational processes; Nature; Negative Reinforcements; neurobiological; Neurobiology; Neuroleptic Agents; Neuroleptic Drugs; Neuroleptics; Neurosciences; novel; operation; Outcome; pathophysiology; Patients; Pb element; Performance; Pharmaceutic Preparations; Pharmaceutical Preparations; Physiopathology; Plant Roots; Positive Reinforcements; Prefrontal Cortex; Process; programs; Programs (PT); Programs [Publication Type]; Psychological reinforcement; Punishment; Reinforcement; Reinforcement (Psychology); Relative; Relative (related person); Research Design; research study; response; reward processing; Rewards; Role; role model; root; Sampling; Schizophrenia; schizophrenic; Schizophrenic Disorders; Series; Short-Term Memory; Side; side effect; Signal Transduction; Signal Transduction Systems; Signaling; Simulate; simulation; Simulation, Computer based; social role; Structure; study design; Study Type; System; System, LOINC Axis 4; Testing; Therapeutic; therapy adverse effect; Tranquilizing Agents, Major; Translating; Translatings; treatment adverse effect; Treatment Side Effects; Universities; Update; virtual simulation; Work; working memory

Project start date: 2008-02-25

Project end date: 2013-01-31

Budget start date: 1-FEB-2011

Budget end date: 31-JAN-2012

PFA/PA: PAR-07-155

5R01MH080066-04 (2011): $602006

QUANTITATIVE BIOMARKER IMAGING FOR EARLY THERAPY RESPONSE ASSESSMENT IN CANCER

M James, Professor
University Of Pittsburgh At Pittsburghcity: Pittsburgh    country: United States (us)

Grant 5U01CA140230-03 from National Cancer Institute

Abstract: There is a vital need for quantitative assessment of cancer therapy response. CT and standard MRI cannot provide information on the molecular, biochemical and physiologic properties of cancer tissues. Therefore, novel quantitative imaging techniques and protocols are needed to reveal biomarkers of molecular events induced by cancer therapy. In particular, early imaging of molecularly targeted pathways predicted to be essential for effective cancer therapy is highly likely to play a key role in patient management in the future. F- 18 FDG-PET can assess the glycolytic response of tumors to chemotherapy and decreases in FDG uptake after the first chemotherapy cycle correlate with better outcome. F-18 FLT PET measures cell proliferation rate, another fundamental process in malignancy. Apoptosis is the primary mechanism of action of most anticancer drugs and can be monitored by the novel PET tracer F-18 ApoSense. In all cancer therapy trials antiangiogenic effect will be measured by DCE MRI in addition to perfusion MRI. Similarly, MRSI can be used to assess choline metabolism, which reflects membrane turnover, through the measurement of total choline and phosphocholine. Finally, triple quantum filtered sodium MRI has been proposed to assess proliferative activity in tumors and changes in cell volume fraction as a result of cell kill. In this project, measures of novel quantitative imaging biomarkers will be combined with patient outcome and tissue biomarkers to develop predictive methodologies for early assessment of response to cancer therapy. The overarching goal of this project will be to standardize PET-CT and MRI protocols, to accurately and reproducibly measure changes in imaging biomarkers during cancer therapy trials, in order to optimize early predictions of subsequent clinical outcomes. Quantitative imaging will be performed in malignant brain tumors with responses to molecularly targeted agents imaged by F-18 ApoSense, F-18 FLT and MRI. Quantitative imaging will also be performed in recurrent or metastatic squamous cell carcinoma of head and neck using novel targeted agents against epidermal growth factor receptor (EGFR) and angiogenesis, including vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR). The type and timing of imaging are aimed at more specific measurements of the effects on the drug target, to provide the earliest indicator of therapeutic response. Development and use of quantitative imaging for early therapy assessment will greatly facilitate patient management, by sparing patients from weeks or months of toxicity and ineffective treatment. RELEVANCE (See instructions) In this project, measures of novel quantitative imaging biomarkers will be combined with cancer patient outcome to develop predictive methodologies for early assessment of response to cancer therapy. Imaging biomarkers of therapy induced early changes in tumor biology will be serially obtained and rigidly quantified. Development and use of quantitative imaging for early therapy assessment will greatly facilitate cancer patient management by sparing patients from weeks or months of toxicity and ineffective treatment

Keywords: angiogenesis; Angiogenesis Inhibitors; Antineoplastic Agents; Apoptosis; base; Biochemical; biomarker; Biometry; Cancer Patient; cancer therapy; cell killing; Cell Proliferation; Cell Volumes; chemotherapy; Choline; Clinical; Clinical Trials Design; Computer software; Data; Data Set; design; Development; Drug Delivery Systems; Early treatment; Epidermal Growth Factor Receptor; Event; Future; Glioblastoma; glucose metabolism; Goals; Head and Neck Cancer; Head and neck structure; Image; Image Analysis; Imaging Techniques; Industry; Instruction; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Malignant neoplasm of brain; Malignant Neoplasms; Measurement; Measures; Membrane; Metabolism; Metastatic Squamous Cell Carcinoma; Methodology; Modality; Modeling; Molecular; Monitor; MRI Scans; novel; Outcome; Pathway interactions; Patients; Perfusion; PET/CT scan; Phosphorylcholine; Physiological; Play; Policies; Positron-Emission Tomography; Principal Investigator; Process; programs; Property; Protocols documentation; quantum; Recurrence; response; Scanning; sharing data; Sodium; Testing; Therapeutic; therapy outcome; Time; Tissues; tool; Toxic effect; Tracer; Treatment Protocols; tumor; Tumor Biology; Universities; uptake; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factors

Project start date: 2009-09-01

Project end date: 2014-07-31

Budget start date: 31-AUG-2011

Budget end date: 31-JUL-2012

PFA/PA: PAR-08-225

5U01CA140230-03 (2011): $491577

BUILDING INTERDISCIPLINARY RESEARCH CAREERS IN WOMEN´S HEALTH

M James, Professor
Magee-women´s Res Inst And Foundationcity: Pittsburgh    country: United States (us)

Grant 5K12HD043441-10 from Office Of The Director, National Institutes Of Health

Abstract: The University of Pittsburgh has a commitment to excellent training in women´s health. The Department of Internal Medicine provides a Women´s Health Residency Track and Fellowship. The Epidemiology Department has led research in women´s health for 20 years and provides a program in the Epidemiology of Women´s Health. In 1992 the Magee-Women´s Research Institute was begun to specifically address research questions relevant to women´s health and catalyzed a further increase in women´s health research and interdepartmental collaboration. Four years ago this culminated in the awarding of the BIRCWH award to the University of Pittsburgh. Through the grant period eight beginning investigators have participated and continue to participate as scholars. The program has attempted to improve women´s health research at the University of Pittsburgh with several strategies. The first has been to provide excellent interdisciplinary research training in women´s health to as many beginning investigators as possible. Scholars funded by the program are encouraged by to obtain alternative K funding and when successful to continue to participate in the BIRCWH career development program. The program has publicized the availability of components of the BIRCWH program to other beginning investigators. In addition, we have recruited many of the research leaders of the University of Pittsburgh to become actively involved in the program through membership in the Advisory Committee. The training program emphasizes interdisciplinary research and exposure to this strategy is provided through projects but also by selecting a group of scholars with diverse research interests and approaches and encouraging their interaction. The Advisory Committee is also made up of investigators with very diverse research interests and strategies. The committee meets with and evaluates both scholar and mentor(s) every 6 months and also provides an interdisciplinary experience for the scholar. The program has been successful with a many publications, grant applications and funding for the scholars. theless, in the next submission we plan modifications. We will formalize team mentoring. We will work with the Center for Minority Health and with a new External Advisory Committee that includes representation from traditionally minority schools to increase minority recruitment. We also establish benchmarks towards extramural funding. With these approaches .we propose to make our program even more successful

Keywords: Address; Advisory Committees; Applications Grants; Award; Benchmarking; career; career development; Collaborations; Epidemiology; experience; Exposure to; Extramural Activities; Fellowship; Funding; Grant; improved; Interdisciplinary Study; interest; Internal Medicine; meetings; Mentors; Minority; minority health; Modification; programs; Publications; Recruitment Activity; Research; Research Institute; Research Personnel; Research Training; Residencies; Schools; Training; Training Programs; Universities; Woman; Women`s Health; Work

Project start date: 2002-09-28

Project end date: 2012-07-31

Budget start date: 1-AUG-2011

Budget end date: 31-JUL-2012

PFA/PA: RFA-OD-06-004

5K12HD043441-10 (2011): $432180

MUSCULAR DYSTROPHY CENTER CORE LABORATORIES

M James, Professor
University Of Minnesota Twin Citiescity: Minneapolis    country: United States (us)

Grant 5P30AR057220-03 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases

Abstract: The University of Minnesota Muscular Dystrophy Center (UMN-MDCenter) has grown in scope and breadth since being organized to improve collaboration in basic, clinical and translational investigation of muscle biology and disease. Success of the MDCenter has recently helped recruit several prominent muscle investigators to Minnesota, capitalizing on existing strength to create what is now a unique multifaceted program focused on understanding muscle and treating muscle disease. To maximize interaction, collaboration and productivity of the expanding MDCenter, we are applying for NIAMS Core Center support with the following specific aims 1) Core B To strengthen and increase accessibility of a research core that provides clinical specimens to muscle investigators, and uses the CLIA-certified muscle biopsy laboratory to characterize human and animal muscle with comprehensive histological and immunostaining methods. 2) Core C To strengthen and expand a repository of murine muscular dystrophy models for muscle investigators, which is coupled with the unique ability of our Center to characterize human and animal muscle force generation at molecular, cellular and whole tissue levels. 3) Core A To provide administrative support for the research cores, and to establish a pilot and feasibility program for new independent investigators, which will strengthen and extend current MDCenter programs for undergraduate, graduate and post-doctoral trainees. 4) To increase institutional, regional and national awareness of UMN-MDCenter and CCMBM to enhance provision of institutional resources commensurate with the potential of this program

Relevance: This Core Center will provide research methods, specimens and support to help University of Minnesota investigators study muscle function and disease more thoroughly and efficiently

Project start date: 2009-08-01

Project end date: 2014-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: RFA-AR-08-003

5P30AR057220-03 (2011): $604000

UNIVERSITY OF MARYLAND SUMMER TRAINING AND RESEARCH (STAR) PROGRAM

M James, Professor
University Of Maryland College Pk Campuscity: College Park    country: United States (us)

Grant 5R25HL092604-04 from National Heart, Lung, And Blood Institute

Abstract: The University of Maryland School of Public Health proposes to implement a Summer Training and Research (STAR) program to provide underrepresented undergraduate students with two consecutive 10-wk summer research and career development programs to enhance their potential to apply for and complete graduate degrees in biomedical and behavioral science relevant to preventing and treating cardiovascular (CV) disease. The aim of the STAR program is to increase diversity in the pool of individuals who ultimately choose careers in biomedical and behavioral research to reduce premature morbidity and mortality from CV and related diseases. Specific objectives are to (1) Increase knowledge of landmark research on the causes and prevention of CV and related diseases, (2) Enhance research skills and experience necessary to be competitive graduate school applicants in CV and related disease research areas, (3) Enhance awareness of the process, practice, and ethics underlying scientific research, (4) Prepare trainees for graduate school and a research career, and (5) Provide quality mentoring. The program will span two consecutive summers, with distinct activities for each summer. Applicants will be recruited locally and nationally through mailings, our Internet web site, visits to undergraduate campuses, and faculty networks. An Admissions Committee consisting of the Program Directors and a minority graduate student will select trainees and match them with faculty mentors. Trainees will be provided with on-site housing and remunerated for their laboratory-based work. The primary efforts of the STAR trainees will focus on laboratory research closely integrated with that of their mentor. The research training component will comprise ~85% of the trainees´ time, with the remaining 15% spent in research-related training and career development. Faculty mentors were selected from those in the School of Public Health with research programs in content areas that are highly relevant to NHLBI. The mix of expertise ranges from laboratory-based bench research to applied community-based research. In addition to direct laboratory experience, trainees will meet weekly to discuss research activities and set weekly goals, meet with current graduate students for dynamic exchanges about graduate studies issues, and discuss classic research articles on cardiovascular disease. Weekly sessions on either Research-Related Training (first year trainees) or Career Development (second year trainees) is included. These sessions are highly interactive and provide trainees with unique perspectives to consider and discuss. A second-year mentor/first- year mentee program is also included to enhance trainee retention over two summers. Trainees will present their research in both oral and poster formats at the end of the summer session. Scientific, social, and cultural activities in the Washington, DC area are planned. Extensive program evaluation will assess how well the program achieves pre-determined benchmarks. Internal and External Advisory Committees will meet to review outcome data and make recommendations to improve and maintain program quality. The University of Maryland School of Public Health proposes to implement a Summer Training and Research (STAR) program to provide underrepresented undergraduate students with two consecutive 10-wk summer research and career development programs to enhance their potential to apply for and complete graduate degrees in biomedical and behavioral science relevant to preventing and treating cardiovascular (CV) disease. The aim of the STAR program is to increase diversity in the pool of individuals who ultimately choose careers in biomedical and behavioral research to reduce premature morbidity and mortality from CV and related diseases. The primary efforts of the STAR trainees will focus on laboratory research closely integrated with that of their mentor. In addition, trainees will meet weekly to discuss research activities and set weekly goals, meet with current graduate students for dynamic exchanges about graduate studies issues, discuss classic research articles on cardiovascular disease, and take part in either Research-Related (first year trainees) or Career Development (second year trainees) Training.(End of )

Keywords: ing; Address; Admission activity; Advisory Committees; African American; Age; Area; Awareness; Bachelor`s Degree; base; Behavioral Research; Behavioral Sciences; Benchmarking; Biomedical Research; Blood Pressure; burden of illness; Cardiovascular Diseases; Cardiovascular system; career; career development; Cessation of life; Commit; Communities; Data; design; Disease; District of Columbia; Education Projects; Educational aspects; Ethics; Ethnicity aspects; experience; Faculty; Fostering; Goals; graduate student; Health; health disparity; Healthy People 2010; Hispanics; Housing; Hypertension; improved; Income; Individual; Institution; Internet; Knowledge; Laboratories; Laboratory Research; Latino; Life; Life Experience; low socioeconomic status; Mails; Maryland; Measures; meetings; member; Mentors; Minority; Mission; Morbidity - disease rate; Mortality Vital Statistics; National Heart, Lung, and Blood Institute; Obesity; Oral; Outcome; Overweight; Population; posters; premature; Prevalence; prevent; Prevention; Process; Program Evaluation; programs; public health medicine (field); Public Health Schools; Race; Recommendation; Recruitment Activity; Research; Research Activity; research and development; Research Personnel; Research Training; Schools; Site; Site Visit; skills; social; Strategic Planning; Students; Time; Training; Universities; Ursidae Family; web site; Work

Project start date: 2008-04-15

Project end date: 2013-03-31

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

PFA/PA: RFA-HL-07-013

5R25HL092604-04 (2011): $89856

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PREDOCTORAL TRAINING IN EXERCISE PHYSIOLOGY AND AGING

M James, Professor
University Of Maryland College Pk Campuscity: College Park    country: United States (us)

Grant 5T32AG000268-13 from National Institute On Aging

Abstract: The University of Maryland College Park Department of Kinesiology exercise biology and aging program has a history of producing well-trained PhDs who complete postdoctoral training and become productive faculty members. We have supported 18 trainees over the first 9 yrs of this program with 12 completing their PhD degrees, 5 still in training, and 1 pursuing an MD/PhD degree in a different discipline (neuroscience). Eleven of the 12 who graduated (92%) have gone on to NIH postdoctoral research fellowships. Seven of these 12 graduates are now in tenure-track academic positions, 1 is in industry, 1 is part-time faculty, and 3 are still in postdoctoral positions. Furthermore, we have graduated 3 African-American women who have gone on to NIH postdoctoral research fellowships at UT-Southwestern, Wake Forest, and Temple. This program provides trainees with the skills necessary for them to complete high-quality dissertation research projects addressing 1) age-related changes in body composition, metabolic, CV, and musculoskeletal function, and CV disease risk factors; 2) exercise training-induced changes in these same parameters; 3) the genetic architecture underlying the baseline measures of these parameters, their changes with aging, and their changes with exercise training interventions; and 4) the mechanisms responsible for these responses in animal and cell culture models using cell and molecular biology methodologies. A substantial enhancement of this renewal Training Program is this translational component, whereby trainees can extend their studies from human physiological studies to investigations in animal models and cell culture to delineate the underlying cell and molecular mechanisms. Trainees´ research can be conducted with Primary, Secondary, and Associate Mentors at closely collaborating laboratories at the University of Maryland School of Medicine Division of Gerontology, Division of Endocrinology, Diabetes, and Nutrition, Department of Physiology, and School of Nursing and Childrens National Medical Center Research Center for Genetic Medicine. Trainees complete coursework in the basic biology of aging; specific knowledge in exercise, CV, musculoskeletal, metabolic, cell, and molecular biology and physiology; genetics; and biostatistics and research design. We propose to continue this program to fund each of 6 predoctoral trainees for 4 yrs to complete their PhD degrees with high-quality coursework and a dissertation research project that will prepare them for an NIH postdoctoral research fellowship and an eventual academic faculty position with a research program focusing on exercise biology and aging. RELEVANCE This program will support 6 trainees/yr who will complete their PhD degrees with high quality research projects. The long-term goal is to prepare trainees to pursue high-level NIH postdoctoral research fellowships and, eventually, academic faculty positions that focus on research and teaching in the prevention of such major age-associated issues as sarcopenia, obesity, diabetes, hypertension, and CV disease

Keywords: Aging; Exercise Physiology; pre-doctoral; Training

Project start date: 1999-05-01

Project end date: 2014-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-06-468

5T32AG000268-13 (2011): $210924

DEVELOPMENT OF NOVEL TOPOISOMERASE AND REPLICATION INITIATOR ASSAYS

M James, Professor
University Of California Berkeleycity: Berkeley    country: United States (us)

Grant 5R01AI091412-02 from National Institute Of Allergy And Infectious Diseases

Abstract: The reliable measurement of enzyme activity is a cornerstone of biochemistry. Biochemical assays are used in nearly all studies of proteins to answer fundamental questions about protein/ligand interactions, catalytic mechanism, and the control of biological processes. Assays also comprise a critical component of high-throughput screens for small molecule agents that disrupt or modulate function. Compounds isolated or re-engineered from such efforts allow researchers to analyze enzyme action in vitro and in vivo, and provide essential therapeutics for the treatment of disease. To serve as a reporter for high-throughput screens, an assay must satisfy stringent technical demands. An inability to meet these requirements - e.g., low cost, high sensitivity and reliability, ease of use, and portability to 96-well or higher formats - can significantly impede the identification of small molecule agonists and antagonists. This problem is particularly acute if the type of reaction catalyzed by a target enzyme is difficult to characterize without the use of classic methods such as gel electrophoresis or radiolabeling. Many proteins responsible for copying and packaging genomic information fall into this class of challenging targets. DNA replication is a defining event in cell proliferation that has long attracted the attention of the basic research sector. Replication factors also have served as valuable targets for inhibitors with beneficial antibiotic, antiviral, or anti-tumorigenic properties. Unfortunately, resistance to and/or toxic side effects from these agents require that new drugs be found. There is likewise a pressing need to develop new small molecule effectors of replication enzymes for probing molecular mechanism. The identification of such agents requires screening chemical libraries against appropriate reporter assays via high-throughput approaches. The goal of this application, developed in synergy with program announcement PA-07-320, is to take advantage of recent structural and conceptual breakthroughs from my lab and the field to design and benchmark a suite of new high-throughput assays for two essential bacterial enzymes type II topoisomerases and the replication initiator, DnaA. We expect that this effort will not only provide much-needed new tools for studying these proteins, but also will pave the way for small molecule screens to identify new probes for biochemical function and novel leads for antibiotic-discovery effort. The pathway to identifying drugs for the treatment of disease requires specialized biochemical assays that report on specific enzymatic activities and their inhibition by clinical agents. This effort will develop several new assays that monitor key reactions carried out by two essential bacterial proteins in support of DNA replication and cell growth. These assays will serve as valuable tools for high-throughput screening programs aimed at identifying new antibiotics for the treatment of acute and chronic infections

Keywords: Acute; Adverse effects; Agonist; Antibiotic Therapy; Antibiotics; Antiviral Agents; ATP Hydrolysis; Attention; Bacteria; Bacterial Proteins; bacterial resistance; base; Basic Science; Benchmarking; Biochemical; Biochemical Reaction; Biochemistry; Biological Assay; Biological Process; catalyst; cell growth; Cell Proliferation; Cell Survival; Cells; Chemicals; Chemistry; Chromosomes; Chronic; Clinical; cost; cytotoxic; design; Development; Dimerization; Disease; DNA; DNA biosynthesis; Drug resistance; Engineering; enzyme activity; Enzymes; Event; falls; Fluorescence; Fluoroquinolones; gel electrophoresis; Gene Expression; Genomics; Goals; Health; high throughput screening; Human; In Vitro; in vivo; Infection; inhibitor/antagonist; insight; interest; Lead; Life; Ligands; Measurement; meetings; melting; Methods; Molecular Probes; Monitor; NIH Program Announcements; novel; Pathway interactions; Pharmaceutical Preparations; Play; portability; Process; programs; Property; protein function; Proteins; public health relevance; Radiolabeled; radiotracer; Reaction; Reader; Replication Initiation; Replication Origin; Reporter; Reporting; Research Personnel; Resistance; Role; Route; Screening procedure; self assembly; Series; small molecule; small molecule libraries; Staging; Structure; Superhelical DNA; System; Therapeutic; Time; tool; Topoisomerase; Topoisomerase II; tumorigenic; Work

Relevance: The pathway to identifying drugs for the treatment of disease requires specialized biochemical assays that report on specific enzymatic activities and their inhibition by clinical agents. This effort will develop several new assays that monitor key reactions carried out by two essential bacterial proteins in support of DNA replication and cell growth. These assays will serve as valuable tools for high-throughput screening programs aimed at identifying new antibiotics for the treatment of acute and chronic infections

Project start date: 2010-06-01

Project end date: 2013-05-31

Budget start date: 1-JUN-2011

Budget end date: 31-MAY-2012

PFA/PA: PA-07-320

5R01AI091412-02 (2011): $372354

SHARED RESOURCES-ANIMAL GENOMICS

M James
Yale Universitycity: New Haven    country: United States (us)

Abstract: The purpose of the Animal Genomics Shared Resource (AGS) is to create (and through cryopreservation, maintain) transgenic and knockout models for human diseases including cancer. The specific objectives are 1) Provide consultation on transgene and targeting construct design; 2) Introduce DNA into zygotes or embryonic stem cells; 3) Create founder animals or chimeras that incorporate the desired, DNA addition or change; and 4) Cryopreserve mouse embryos for cost-effective, long-term storage of genetically engineered mice. The Transgenic mouse service was created in 1989. The ability to create knockout mice was added in 1997. Cryopreservation of mouse embryos has been performed since 2000. The creation of novel mouse strains, either by transgenesis or homologous recombination, allows many members of the Yale Cancer Center (YCC), to determine the normal function of genes and how perturbation of that function can result in aberrant development or diseases such as cancer. Investigators can use AGS to create direct analogs of mutations involved in human disease, thus providing models that can be studied in a way that human subjects cannot. The ability to create such induced mutants allows investigators to effectively use mouse genetics to dissect and analyze complicated developmental pathways, biochemical processes, and biomolecular interactions in a mammal to a level previously unattainable before such technology existed. This enables researchers to investigate biological processes at a far deeper level than in vitro methods such as biochemistry or cell culture will allow, and through the appearance of novel phenotypes resulting from induced mutations, discover new relationships between these processes and pathways. Cryopreservation of mouse strains allows investigators to more efficiently utilize their financial resources in research while maintaining valuable strains for future use. The YCC AGS produces 40-60 transgenic mouse strains/year, and creates 15-20 new targeted mutant mouse models/year. Animal Genomics Services has been a shared resource of the YCC since 1989. AGS moved into new space designed specifically for their use with the completion of The Anylan Center building in 2002. Twenty-eight YCC investigators used AGS in 2005 for a total of approx. 62% of all usage of the resource. All YCC usage during 2005 was from peer-reviewed scientists from 7 of 8 YCC Research Programs

Keywords: analog; animal resource; Animals; Appearance; Biochemical Process; Biochemistry; Biological Process; Cell Culture Techniques; Chimera organism; Comprehensive Cancer Center; Consultations; cost effective; Cryopreservation; design; design and construction; Development; Disease; DNA; Embryo; embryonic stem cell; Future; gene function; Gene Transfer Techniques; Genetic; Genetically Engineered Mouse; Genomics; Genomics Shared Resource; homologous recombination; human disease; human subject; In Vitro; Induced Mutation; Knock-out; Knockout Mice; Malignant Neoplasms; Mammals; member; Methods; Modeling; Mouse Strains; Mus; mutant; mutant mouse model; Mutation; novel; Pathway interactions; Peer Review; Phenotype; Process; programs; Research; Research Personnel; Resource Sharing; Resources; Scientist; Services; Technology; Transgenes; Transgenic Mice; Transgenic Organisms; Yale Cancer Center; zygote

Budget start date: 1-AUG-2011

Budget end date: 31-JUL-2012

5P30CA016359-33_0009 (2011): $182742

ESTABLISHING AN EXPLORATORY NCMHD RESEARCH CENTER OF EXCELLENCE IN ARKANSAS

M James
University Of Arkansas Med Scis Ltl Rockcity: Little Rock    country: United States (us)

Grant 5P20MD002329-05 from National Institute On Minority Health And Health Disparities

Abstract: The proposed theme is to develop research to improve access to quality prevention and healthcare programs for racial and ethnic minorities with a goal of eliminating health disparities. The planned focus is on chronic disease disparities, based on the major sources of disparities, with initial emphasis on CVD, cancer, and associated risk factors of obesity, diabetes, tobacco use, physical inactivity, and risky sexual behavior. Initial proposed research projects include 1 Full and 6 Pilot Projects. We propose to align the Center with major sources of identified health disparities through an evidence-based process. Finally, we propose to develop 3 cores to support the Center an Administrative Core to oversee day-to-day activities and establish policy for the proposed Center and interface with National and Local Advisory Committees; a Research Core to develop recommendations for Center directions based on an evidence-based approach to priority and heuristic theoretical model evolution; and an Education/Training Core to promote minority student recruitment into careers in health disparities research through a combined BA/MPH degree program in concert with Arkansas´ three HBCUs. The University of Arkansas for Medical Sciences (UAMS) Fay W. Boozman College of Public Health (COPH) was established in July 2001, after a public referendum led to the creation of a new College based on a carefully, strategically planned Vision, Mission, and Values, emphasizing the health and well-being of Arkansans and eliminating health disparities. The COPH, the lead in this application, is thus based on the very scientific focus of this application - the elimination of health disparities. Our commitment to the elimination of health disparities is evidenced by a number of structures and initiatives as described in our application. Nowhere is this organizational commitment more evident than with the Pi/Director for the proposed Center, Dr. Jim Raczynski, who is Founding Dean and former Founding Director for 9 years of the DAB Center for Health Promotion, one of CDC´s Prevention Research Centers, focusing on primary prevention among African Americans in rural Alabama. As Dean, he acts as the chief executive/academic officer of the College, enabling him to ensure the adequacy of financial, space, and administrative support resources to ensure the success of the proposed Exploratory NCMHD in Arkansas, a commitment that is confirmed by by Chancellor I. Dodd Wilson, the CEO of UAMS

Project start date: 2007-09-30

Project end date: 2012-05-31

Budget start date: 1-JUN-2011

Budget end date: 31-MAY-2012

PFA/PA: RFA-MD-06-003

5P20MD002329-05 (2011): $1176653

5P20MD002329-04 (2010): $1063835

ANALYTIC METHODS FOR HIV TREATMENT AND CO-FACTOR EFFECTS

M James
Harvard University (sch Of Public Hlth)city: Boston    country: United States (us)

Grant 5R37AI032475-20 from National Institute Of Allergy And Infectious Diseases

Abstract: The principal aim of this proposal is to further development of new methods for analyzing observational data bases and randomized trials of HIV-infected persons and the application of these methods to data obtained in randomized and observational studies in an attempt to help answer important open substantive questions concerning the treatment and course of HlV-related disease. The proposed approaches are based either on (i) the estimation of new classes of causal models which include structural nested models, marginal structural models (MSMs), direct effect structural nested models, continuous time structural nested models, and optimail regime structural models (SNMs). Many of the new methods are fundamentally "epidemiologic" in that they require data on time-dependent confounding factors, that is, risk factors for outcomes that also predict subsequent treatment with the drug or cofactor under study. In particular, we plan to further develop optimal regime SNMs and dynamic MSMs to help detemnine the optimal times to start HAART therapy and to change HAART regimens as a function of a subject´s CD4 count, HIV RNA, clinical history, and, where available, results of genot^lc or phenotypic resistance testing. Our methods will be developed with the goal of directing analyzes and reanalyzes, with collaborators, of data from the HIV Causal Colioboration at HSPH . the Multicenler AIDS Cohort Study, The Women´s Interagency HIV Study, The Swiss HIV Cohort Study, The Study of The Consequences of Protease Inhibitor Era (SCOPE), Pediatric Late Outcomes Protocol (PACTG 219) and the ALLRT study. RELEVANCE (See instructions) Observational methods are used to answer pressing causal questions that cannot be or have not yet been studied in randomized trials. In particular we are developing methods that are the best available to determine the optimal CD4 and HIV RNA levels at which to initiate HAAART therapy in HIV infected subjects and the optimal time to change therapy once resistance to a initial HAART regime has developed

Keywords: Acquired Immunodeficiency Syndrome; Algorithms; Back; base; case control; CCR5 gene; CD4 Lymphocyte Count; Childhood; Clinical; cofactor; cohort; Cohort Studies; Computer software; Computing Methodologies; Confidence Intervals; cost; Cox Proportional Hazards Models; Data; data modeling; Databases; Derivation procedure; design; Development; Disease; Disease Progression; dosage; Epidemic; Epidemiology; Equation; Goals; Graph; Highly Active Antiretroviral Therapy; HIV; Hospitals; Infection; Instruction; Intervention; Investigation; Left; Maps; Markov Chains; Methodology; Methods; Microcomputers; Modeling; Needle Sharing; novel; Observational Study; Outcome; Patients; Persons; Pharmaceutical Preparations; Physicians; Prevalence; Protease Inhibitor; Protocols documentation; public health medicine (field); Randomized; Randomized Controlled Trials; randomized trial; Recording of previous events; Regimen; Research Personnel; Resistance; response; Risk Behaviors; Risk Factors; RNA; Sampling; Sex Behavior; Solutions; Staging; Stochastic Processes; Structural Models; Surveys; Testing; theories; Time; user-friendly; Vaccination; vaccine efficacy; Validation; Viral; Woman; Work

Project start date: 1992-08-01

Project end date: 2015-03-31

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5R37AI032475-20 (2011): $641622

GENOME VARIATION UNDERLYING ALCOHOL ACTION

M James, Professor
University Of Colorado Denvercity: Aurora    country: United States (us)

Grant 5R01AA011853-13 from National Institute On Alcohol Abuse And Alcoholism

Abstract: The primary goal of this application is to identify genes and other genomic features that underlie the action of alcohol. While we have made considerable progress toward this objective through the use mouse quantitative trait loci (QTL) mapping, gene expression profiling and comparative DNA sequencing, these studies were limited and complicated by two newly discovered factors related to the analysis of complex traits 1) mammalian genomic variation, including copy number variation (CNV), is much more prevalent than previously expected , and 2) mouse genome sequence contains many gaps and complex, duplication-rich regions that are likely to be misassembled. We plan to apply two recently developed genomic strategies that circumvent many of these limitations. The first strategy will be to carry out very high density array-based comparative genomic hybridization (aCGH) of several mouse strains (ILS, ISS, LXS RIs and HS4) known to differ in alcohol-related phenotypes, and identify CNVs that are contributing to differences in alcohol action between these strains. CNVs will be independently confirmed, CNV boundaries fine-mapped using custom aCGH arrays, and genes within the CNV identified and characterized. To our knowledge this study will be the first genome-wide investigation of the role of CNVs in alcohol action. The second strategy will be to obtain and compare the complete DNA sequences of four QTL regions (Lores1, 2, 4, and 5) between strains that have been selected for differential sensitivity to alcohol (ILS and ISS). This will be carried out by array-capture of QTL- specific DNAs from each strain, followed by ultra-high throughput DNA sequencing of the complete QTL interval using next generation DNA sequencing technology. This study will identify all DNA sequence variations within each Lore QTL and, as such, will represent the most complete assessment of genomic variation with these alcohol-related QTLs. The proposed experiments have the potential to identify important new genes and pathways that underlie the action of alcohol in mammalian systems. Such new insights could lead to improved strategies for the treatment and prevention of alcoholism and alcohol abuse

Keywords: Address; Alcohol abuse; alcohol sensitivity; alcoholism prevention; Alcohols; base; comparative; comparative genomic hybridization; Complex; Computer software; Copy Number Polymorphism; Custom; density; design; Detection; DNA; DNA Microarray Chip; DNA Sequence; Elements; Foundations; Gene Expression Profiling; Genes; Genome; genome sequencing; genome-wide; Genomics; Goals; Health; Human; Hybridization Array; improved; Inbred Strain; Individual; insight; instrument; Investigation; Lead; mammalian genome; Maps; Methods; mouse genome; Mouse Strains; Mus; next generation; Oligonucleotides; Pathway interactions; Phenotype; Quantitative Trait Loci; research study; Resolution; Role; Sequence Analysis; Slide; Solid; structural genomics; Surveys; System; Technology; trait; treatment strategy; Variant; Variation (Genetics); Work

Relevance: The proposed experiments have the potential to identify important new genes and pathways that underlie the action of alcohol in mammalian systems. Such new insights could lead to improved strategies for the treatment and prevention of alcoholism and alcohol abuse

Project start date: 1998-05-01

Project end date: 2014-05-31

Budget start date: 1-JUN-2011

Budget end date: 31-MAY-2012

PFA/PA: PA-07-070

5R01AA011853-13 (2011): $343241

RESEARCH TRAINING PROGRAM IN THE BEHAVIORAL AND BIOMEDICAL SCIENCES

M James, Professor And Vice Chair
West Virginia Universitycity: Morgantown    country: United States (us)

Grant 5T32GM081741-04 from National Institute Of General Medical Sciences

Abstract: The overall objective of this T32 Predoctoral Training Grant is to provide rigorous research training in the behavioral and biomedical sciences to students planning careers as independent investigators. The goal is to develop the next generation of scientists with the background and skills to conduct behavioral research utilizing a wide range of experimental approaches and incorporating biochemical, molecular, and genetic analyses. The research training program in Behavioral and Biomedical Sciences (BBS) engages experienced faculty preceptors and builds upon established Biomedical, Psychology, and Public Health Ph.D. programs at WVU. We have recruited outstanding, NRSA-eligible students, and have been providing them with high quality training in behavioral and biomedical research. T32 support will allow us to formalize this training program and increase the number of students we can accept into this value-added training initiative. The BBS Steering Committee appoints students to this T32 based on merit - considering their academic qualifications, interest in behavioral research, and commitment to interdisciplinary research training. Focused behavioral and biomedical research training is achieved by 1) supervised research under the direction of a BBS faculty preceptor; 2) BBS-specific core courses - Integrated Analysis of Brain Structure and Function, Experimental Analysis of Behavior, Biostatistics and Bioinformatics, and Epidemiology; 3) BBS Journal Club and Seminar programs; and 4) Research Practicum, which provides students with additional, hands-on research experience through rotations with faculty preceptors engaged in behavioral research utilizing approaches and methodologies different from their own. Student progress is evaluated by the Director and Steering Committee, not only while in the program, but on a regular basis following graduation. This permits measurement of trainees´ research productivity, career trajectories, and accomplishments; it also provides guidance for improving the program for current and future trainees. The success of the BBS program in meeting its training goals is evaluated by Internal and External Advisory Committees, which examine student recruitment and retention, quality of training, and success of graduates in developing as independent scientists addressing complex behavioral and biomedical problems of clinical and public health importance

Keywords: Behavioral; Research Training; Science; Training Programs

Project start date: 2008-07-01

Project end date: 2013-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PAR-06-503

5T32GM081741-04 (2011): $157184

OSTEOPOROTIC FRACTURES IN MEN - MROS RENEWAL - BIRMINGHAM

M James, Associate Professor Of Medicine
University Of Alabama At Birminghamcity: Birmingham    country: United States (us)

Grant 5U01AR045632-13 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases

Abstract: The Osteoporotic Fractures in Men (MrOS) study was formed primarily to quantify the determinants of fracture in men. Importantly, the MrOS cohort also provides a seminal opportunity to study men as they progress through a critical period of life in which problems of aging remain poorly understood. In 2000-2002, 5995 community-dwelling men ages 65 years and older (mean age 72 years at baseline) were enrolled from 6 diverse US communities. After 5 years of follow-up, participant retention is excellent (99% of those alive remain active). We propose a new clinic visit and continued follow-up of the cohort to expand our understanding of risk factors for falls, fractures (particularly hip fracture) and other consequences of aging. At baseline, both areal (from DXA) and volumetric (from QCT) skeletal assessments were obtained. We propose to repeat the same assessments in the planned visit to identify the densitometric and biomechanical risk factors for hip fracture, as well as to characterize bone geometry changes that underlie skeletal fragility. Further, we will use the QCT scans to quantify femoral strength by finite element modeling (FEM), and assess the usefulness of FEM for fracture prediction. Additional follow-up of the cohort and repeat measures of muscle strength (grip strength, leg power), physical performance (gait speed, chair stands, and narrow walk), and self-reported physical activity will enable us to establish the extent and nature of change in activity and physical performance, identify biological predictors of these changes, and clarify the possibly joint effects of physical activity and physical performance on fracture risk. To objectively quantify physical activity we propose to obtain accelerometry measures at the new clinic visit. Using serum archived at baseline, we will test the hypothesis that renal function, vitamin D, parathyroid hormone have important effects on skeletal health, physical function and fracture risk, and will determine if lower sex steroid levels increase men´s risk of skeletal deterioration and fracture, decline in physical function, and deterioration in quality of life. Combined with the considerable data and biologic specimens already collected, additional measures and extended follow-up in the MrOS cohort allows us to expand the understanding of hip fractures, the most devastating type of fracture in men, as well as address other health issues of compelling public health and clinical importance to older US men

Keywords: Address; Age; Aging; Archives; Biological; Biomechanics; bone geometry; Clinic Visits; Clinical; cohort; Communities; critical period; Data; Deterioration; Dual-Energy X-Ray Absorptiometry; Elements; Enrollment; falls; follow-up; Fracture; Gait; Gonadal Steroid Hormones; grasp; Health; Hip Fractures; human old age (65+); human PTH protein; Joints; Leg; Life; Measures; men; Modeling; muscle strength; Nature; osteoporosis with pathological fracture; Participant; Patient Self-Report; Performance; Physical activity; Physical Function; public health medicine (field); Quality of life; Renal function; Risk; Risk Factors; Scanning; Seminal; Serum; skeletal; Specimen; Speed (motion); Testing; Visit; Vitamin D; Walking

Project start date: 1999-09-30

Project end date: 2012-07-31

Budget start date: 1-AUG-2011

Budget end date: 31-JUL-2012

5U01AR045632-13 (2011): $214335

OFFICE OF CLINICAL RESEARCH

M James, Professor
Dartmouth Collegecity: Hanover    country: United States (us)

Abstract: The Office of Clinical Research (OCR) coordinates the review, initiation, conduct, and safety monitoring of clinical trials at the Morris Cotton Cancer Center. The OCR now functions as a branch of the Clinical Trials Office (CTO) of Dartmouth Medical School. The OCR has recently undergone a dramatic expansion in order to better accommodate clinical research needs within the Cancer Center. Components of the OCR are (1) Administrative staff, including the Medical Director, the Administrative Director, the Regulatory Compliance Officer, and the Clinical Operations Coordinator; (2) Research Support staff¿a research pharmacist, a pharmacy technician, six research RNs, and 24 clinical research assistants/coordinators; (3) interface with the Protocol Review and Monitoring System, consisting of the Clinical Cancer Research Committee (CCRC) and the Safety and Data Monitoring Committee (SDMC); (4) the institutionally funded Dartmouth-Hitchcock Tumor Registry, staffed by three registrars; and (5) a Clinical Trials Management System (Velos eResearch). The OCR maintains local Dartmouth Medical School investigator-initiated studies, Dartmouth investigatorinitiated studies relying on a consortia of working group enrolling sites, cooperative group protocols, and industrial trials. It is responsible for assisting in formatting protocols, writing consent forms, maintaining Institutional Review Board (CPHS) and SDMC records, developing budgets, assessing impact on institutional resources, activating protocols, assessing eligibility, developing data collection forms and clinical trials databases, and collecting and managing data. A commitment to research and clinical trial enrollment support has been integrated into the current expansion of the Dartmouth-Hitchcock Medical Center into a regional network of clinical facilities. Prioritization of clinical trials is determined within the 11 NCCC Clinical Oncology Groups, with approval by the CCRC. The OCR acts as a checkpoint prior to activation of protocols to ensure that all appropriate administrative aspects of trials are complete. Chargeback policies of the OCR accurately reflect usage and are based on patient accrual to reviewed and approved protocols. For the CCSG fiscal year ending November 30, 2007, the OCR supported 215 active trials. These trials represented service to 32 different Pis. Support of these studies during this period included the new registration of 484 patients and follow-up for 1,876 patients previously enrolled. For the most recent reporting period for our tumor registry in 2006, we entered 487 of 2,582 (1990) patients on therapeutic trials. Throughout this award period, approximately 40% of patients entered onto intervention trials were accrued to institutional trials

Keywords: Address; Adherence (attribute); Adverse event; Agreement; anticancer research; Award; base; Breast; Budgets; Calendar; Cancer Center; Cancer Center Support Grant; Case Report Form; Clinical; Clinical Nursing Research; Clinical Oncology; Clinical Protocols; Clinical Research; Clinical Trials; Clinical Trials Data Monitoring Committees; Clinical Trials Database; Collaborations; computerized; Conduct Clinical Trials; Consent; Consent Forms; Contracts; Data; Data Collection; data management; Data Reporting; Databases; Development; Disease; Disorder by Site; Documentation; Educational aspects; Eligibility Determination; Enrollment; Ensure; Evaluation; follow-up; Funding; Gossypium; Grant; Gynecologic Oncology; Head and neck structure; Health system; Housing; human subject; Informed Consent; Intervention Trial; Investigational Drugs; leukemia/lymphoma; Lung; Maintenance; Medical center; medical schools; meetings; melanoma; member; Monitor; Monitoring Clinical Trials; New England; Nursing Research; Oncology Group; operation; Patients; Peer Review; Pharmaceutical Preparations; Pharmacists; Pharmacy facility; Physician Executives; Policies; Positioning Attribute; Principal Investigator; programs; Prostate; Protocol Compliance; protocol development; Protocols documentation; quality assurance; Records; Recruitment Activity; Regulation; Reporting; Research; Research Contracts; Research Ethics Committees; Research Personnel; Research Support; Resources; Rural Hospitals; Safety; safety study; Screening procedure; Services; Site; Specialist; Standardization; Supervision; System; Technology; Therapy Clinical Trials; Training; Training Support; tumor registry; Update; Work; working group; Writing

Budget start date: 1-DEC-2010

Budget end date: 30-NOV-2011

5P30CA023108-33_5691 (2011): $202036

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MOLECULAR BIOPHYSICS TRAINING GRANT

M James
Harvard University (medical School)city: Boston    country: United States (us)

Grant 5T32GM008313-23 from National Institute Of General Medical Sciences

Abstract: The Molecular Biophysics Training Grant at Harvard supports a predoctoral training program focused at the interface of the physical and biological sciences. The goal of the program is to provide students with strong undergraduate backgrounds in quantitative sciences (especially physics and mathematics) with broad training in the biophysical, chemical and molecular concepts and techniques that are required to address outstanding problems in biology and biomedical sciences. The training program links a highly interactive group of 44 faculty members from three departments in Harvard´s Faculty of Arts and Sciences, six departments at Harvard Medical School, and four affiliated hospitals. The training program offers a flexible curriculum drawn from courses offered at Harvard, Harvard Medical School, and MIT, and offers research opportunities in a variety of disciplines relevant to molecular biophysics with particular strengths in the areas of structural biology, computational biology, neuroscience, and imaging. In addition to coursework and research activities, the training program sponsors seminars and guest lectures; a student run, student research seminar series; a yearly retreat featuring a poster session and student and faculty talks in the fall semester; a poster session featuring research of program students; a mini-symposium featuring talks by program faculty during the Biophysics Program recruiting weekend in the spring semester; and social events for all trainees. Over the past 19 years, this training program has helped foster a number of new initiatives in graduate training, and has been remarkably successful in promoting collaborative research among its faculty and interdisciplinary training for its students. In this competitive renewal we request support for 16 training slots for students who are affiliated with Harvard´s Biophysics Program or who are jointly affiliated with the Harvard Biophysics Program and Medical Engineering and Medical Physics (MEMP) Ph.D. program in the joint Harvard/MIT Health Sciences and Technology initiative (HST). Students will be preferentially funded in their first and second year of graduate studies. This training program offers interdisciplinary training in approaches to explore the structure, function and interactions of key macromolecules of the cell, the control of cellular processes at the molecular and genome-wide levels, the development of approaches to imaging and modeling these processes, and the use of this information to diagnose, understand, prevent or cure human diseases

Keywords: Biophysics; Grant; Molecular; Training

Project start date: 1989-07-01

Project end date: 2014-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

5T32GM008313-23 (2011): $618632

CORRELATIVE CRYO-MICROSCOPY: A NEW APPROACH FOR CHARACTERIZING THE STRUCTURE AND

M James
Harvard University (medical School)city: Boston    country: United States (us)

Grant 5R01GM085578-04 from National Institute Of General Medical Sciences

Abstract: The macromolecular machines of cells pose structural questions that span over six orders of magnitude in scale from the tenths of nanometers (atoms) to tens of microns (cells). This large range of scale exceeds the capabilities of any one technique. As a result, structural biologists have increasingly turned to the use of hybrid approaches. For example, the combination of x-ray crystallography and cryoelectron microscopy has been widely used to probe the structure and mechanics of large complexes (e.g. viruses, ribosomes, proteosomes, chaperones, actin filaments and microtubules) in vitro at the near atomic level. In this project we will explore technologies to link these first generation hybrid techniques, single molecule optical microscopy, and cryoelectron tomography to characterize the structure and function of macromolecular complexes in the context of the whole cell. The technologies that will be developed include the use of optical imaging methods to characterize the functional state of macromolecular complexes in cells imbedded in vitreous ice and determine their location with very high precision (tens of nm), development of a cold stage that will permit the optical positional mapping to be transferred to the electron microscope, development of computational tools to locate, identify, and determine the structures of macromolecular complexes in cryoelectron tomographic reconstructions, and refinement of computational tools to match these structures with higher resolution structures derived from conventional in vitro methods. In our initial studies we will use the new technologies to characterize the cell entry pathway of poliovirus. This model was chosen because of its biological relevance (the cell entry mechanisms of nveloped viruses are poorly understood) and because structures of several relevant cell entry intermediates in vitro are already known. The techniques developed will be generally applicable to a wide range of macromolecular complexes. In this project will develop technologies that will combine fluorescence optical microscopy and cryoelectron tomography to characterize the structure and function of intracellular macromolecular assemblies at the molecular level in situ. The technology will be relevant to a wide range of intracellular structures, and will add significantly to our basic understanding of the structure and function of "cellular machines."

Keywords: Biological; Cell physiology; Cells; Complex; computerized tools; Cryoelectron Microscopy; Crystallography; Development; Electron Microscope; Fluorescence; Generations; Human poliovirus; Hybrids; Ice; imaging modality; In Situ; In Vitro; Link; Location; macromolecular assembly; Macromolecular Complexes; Maps; Mechanics; Methods; Microfilaments; Microscopy; Microtubules; Modeling; Molecular; Molecular Chaperones; multicatalytic endopeptidase complex; nanometer; new technology; novel strategies; optic imaging; Optics; Pathway interactions; Polioviruses; reconstruction; Resolution; Ribosomes; single molecule; Staging; Structural Biologist; Structure; Subcellular structure; Techniques; Technology; tomography; Virus

Project start date: 2008-08-01

Project end date: 2012-07-31

Budget start date: 1-AUG-2011

Budget end date: 31-JUL-2012

PFA/PA: RFA-GM-08-002

5R01GM085578-04 (2011): $332254

MOLECULAR GENETICS OF HSV DNA POLYMERASE GENE

M James, Pd/pi
Harvard University (medical School)city: Boston    country: United States (us)

Grant 2R01AI019838-26 from National Institute Of Allergy And Infectious Diseases

Abstract: The long-term objective of this research is a detailed understanding of herpesvirus DNA polymerases and drugs that target them. These enzymes, which include a catalytic subunit (Pol) and an accessory subunit that stimulates long-chain DNA synthesis, are both prototype ?-like DNA polymerases and excellent targets for antiviral drugs. This latter property is especially health-related, as new drugs are needed for treatment of herpesvirus infections. In this application, unanswered questions regarding accessory subunits, catalytic subunits, and drugs that target these proteins and their interaction are addressed. Specific aim 1 is to investigate the unusual and different manners by which the accessory subunits, such as herpes simplex virus (HSV) UL42 and human cytomegalovirus (HCMV) UL44, interact with DNA so that they bind tightly, yet diffuse linearly along the DNA to permit processive DNA synthesis. Single-molecule approaches will be used to analyze how these proteins move on DNA, particularly whether wild type UL42 or a tight-binding mutant necessarily moves helically or can, for example, move along one side of the helix. The force required to move these subunits will be compared with the force required to stop or slow the catalytic subunits. X-ray crystallography will be used to understand the molecular details of the protein-DNA interaction. Specific aim 2 is to investigate the roles of structural domains of the catalytic subunits that are N-terminal to the thumb, palm, and fingers domains in terms of enzymatic functions, binding to the base excision repair (BER) enzyme uracil DNA glycosylase (UNG), viral replication, and mechanisms of antiviral drug resistance. Two structural domains, pre-NH2 and NH2, have been observed in the crystal structure of HSV Pol, but their roles in enzyme function and viral replication are unknown. To address these questions, mutant enzymes will be engineered and assayed for relevant biochemical activities. Mutant viruses will be engineered and assayed for viral replication both in cells and in a mouse model. The mechanisms by which mutant HCMV Pols with substitutions in their 3´-5´ exonuclease domain resist ganciclovir (GCV) action will be investigated using enzymological analyses. Specific aim 3 is to discover new compounds that inhibit the interaction of HCMV Pol and UL44, and HCMV replication, using a structure-based approach. The importance of the interaction between UL44 and another replication protein, UL84, for viral replication, and whether it can be exploited as a drug target, will be investigated using a combination of biochemical, molecular genetic, and cell biological approaches, including efforts to develop a new technique to map protein-protein interactions. Should the UL44-UL84 interaction look promising as a drug target, random screening for compounds that inhibit this interaction will be undertaken. Herpesviruses cause widespread disease in the population at large and severe disease in people with impaired immunity. There is considerable need for new drugs to combat these viruses. The research proposed should not only provide information that could aid in drug discovery and understanding how viruses become resistant to current drugs, but aims directly to discover new anti-herpesvirus drugs

Keywords: Acyclovir; Address; Antiviral Agents; base; Base Excision Repairs; Binding (Molecular Function); Biochemical; Biological; Biological Assay; Biology; Catalytic Domain; Cells; combat; Cytomegalovirus; Development; Diffuse; Diffusion; Disease; DNA; DNA Binding; DNA biosynthesis; DNA-Directed DNA Polymerase; DNA-Protein Interaction; Drug Delivery Systems; drug development; drug discovery; Drug resistance; Engineering; Ensure; Enzymes; Exonuclease; Fingers; Ganciclovir; Genes; Grant; Health; Herpesviridae; Herpesviridae Infections; high throughput screening; Homologous Gene; Human; human DNA; Humulus; Immunity; improved; in vivo; inhibitor/antagonist; interdisciplinary approach; interest; Lead; Learning; Lyase; Maps; Methods; Molecular; Molecular Genetics; mouse model; mutant; Mutation; N-terminal; Pharmaceutical Preparations; Phosphodiesterase I; Polymerase; Polymerase Gene; Population; Property; Protein Binding; Protein-Protein Interaction Map; Proteins; prototype; public health relevance; repair enzyme; Research; Resistance; resistance mutation; Role; Screening procedure; Side; Simplexvirus; Simplexvirus DNA polymerase; single molecule; Slide; Structure; Techniques; Thumb structure; uracil-DNA glycosylase; Viral; Viral Drug Resistance; Viral Physiology; Virus; Virus Replication; Widespread Disease; X-Ray Crystallography

Project start date: 1983-04-01

Project end date: 2016-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-10-067

2R01AI019838-26 (2011): $507375

CYTOSKELETAL INTERACTIONS OF DYSTROPHIN

M James, Professor
University Of Minnesota Twin Citiescity: Minneapolis    country: United States (us)

Grant 5R01AR042423-17 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases

Abstract: The long-term objective of this project is to determine the function(s) of dystrophin in striated muscle in order to understand how its absence or abnormality leads to the pathologies observed in Duchenne (DMD) and Becker (BMD) muscular dystrophies. During the most recent project period, we uniquely employed the baculovirus expression system and an array of biochemical/biophysical assays to define the actin binding properties of dystrophin and its homologue utrophin. We also demonstrated that dystrophin functions to organize the microtubule lattice of skeletal muscle through a direct binding interaction. Finally, we demonstrated that a baculovirus-expressed TAT-micro-utrophin construct shows promise as a novel protein replacement therapy for DMD. Based on these published results and exciting new preliminary data, we propose three new specific aims to 1) test the hypothesis that disease-causing missense mutations in dystrophin cause a loss of function through protein aggregation, 2) determine the functional importance of microtubule lattice organization in skeletal muscle, and 3) reinvestigate functional necessity for the dystrophin carboxyl-terminal domain in skeletal muscle. The project will employ biochemical/biophysical methods to characterize a variety of dystrophin and utrophin protein constructs in combination with complementary experiments in isolated cells and transgenic mice. The proposed studies will address several fundamental questions about the function of dystrophin in normal skeletal muscle and how its absence or abnormality causes human diseases of skeletal muscle. Duchenne muscular dystrophy (DMD) afflicts 1 in 3,500 live-born males and is typically lethal by the third decade. No effective therapies are currently available for DMD. Dystrophin is the protein missing in patients with DMD, but its function in muscle is not fully understood. Through rigorous biochemical/biophysical characterization of full-length dystrophin and utrophin in combination with complementary experiments in isolated cells and transgenic mice, the proposed studies will elucidate the functions of dystrophin in normal muscle and how its absence causes muscular dystrophy. Thus, this project is highly relevant to understanding the pathological mechanism of Duchenne muscular dystrophy and for the development of effective therapies

Keywords: Actins; Address; Affinity; Baculovirus Expression System; Baculoviruses; base; Binding (Molecular Function); Biochemical; Biological Assay; Buffers; calponin; Cells; Data; Dependence; Development; Dilated Cardiomyopathy; Disease; Dystrophin; effective therapy; enzyme replacement therapy; Exons; Fibroblasts; flexibility; Homologous Gene; human disease; In Vitro; in vivo; Inbred mdx Mice; Induced Mutation; knockout animal; Label; Length; Life; loss of function; male; Masks; Measures; Methods; Microfilaments; Microtubule-Associated Proteins; Microtubules; Missense Mutation; Molecular; Morphology; Mus; Muscle; Muscle Fibers; Muscle function; Muscular Dystrophies; Muscular Dystrophy, Becker; Muscular Dystrophy, Duchenne; mutant; Mutation; novel; overexpression; Pathology; Patients; Phenotype; Point Mutation; Property; protein aggregation; Protein Isoforms; Proteins; Proteomics; public health relevance; Publishing; Recombinant Proteins; research study; Rods (Retina); Role; Skeletal muscle structure; Striated Muscles; Structure; System; Temperature; Testing; Time; Transgenes; Transgenic Mice; Transgenic Organisms; Utrophin

Relevance: 7. Duchenne muscular dystrophy (DMD) afflicts 1 in 3,500 live-born males and is typically lethal by the third decade. No effective therapies are currently available for DMD. Dystrophin is the protein missing in patients with DMD, but its function in muscle is not fully understood. Through rigorous biochemical/biophysical characterization of full-length dystrophin and utrophin in combination with complementary experiments in isolated cells and transgenic mice, the proposed studies will elucidate the functions of dystrophin in normal muscle and how its absence causes muscular dystrophy. Thus, this project is highly relevant to understanding the pathological mechanism of Duchenne muscular dystrophy and for the development of effective therapies

Project start date: 1994-07-15

Project end date: 2015-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PA-07-070

5R01AR042423-17 (2011): $352864

INTERACTIONS BETWEEN CYTOTOXIC AND ANTIANGIOGENIC DRUGS

M James, Professor
Mount Sinai School Of Medicinecity: New York    country: United States (us)

Grant 2R01CA072937-09 from National Cancer Institute

Abstract: The growth and invasiveness of solid tumors is highly dependent on angiogenesis or the processes forming new tumor blood vessels. Accordingly the development of antiangiogenic drugs is of considerable importance, yet recent clinical trials have demonstrated that early favorable patient responses are not durable, which has been attributed to drug resistance. The complex and diverse nature of the drug resistance mechanisms calls for a systematic approach to define new treatment paradigms to alleviate resistance to angiogenesis inhibitors. The overall objective of the project is to provide a preclinical foundation to design multidrug combination regimens that will overcome resistance to angiogenesis inhibitors and further ensure that coadministered cytotoxic drugs will reach tumors in sufficient amounts. To accomplish this objective, three Aims are proposed that describe a series of pharmacokinetic (PK) and pharmacodynamic (PD) investigations based on the properties of the drugs in tumors. Aim 1 studies will derive antiangiogenic drug resistant brain tumors in vivo and compare the tumor accumulation of the cytotoxic drug, temozolomide (TMZ) in sensitive and resistant tumors. The expression of genes and proteins relevant to angiogenesis will be monitored to create a resistance profile. Initial Aim 2 studies, again utilizing drug resistant tumors, will evaluate multitargeted drug combinations that interfere with angiogenesis by inhibiting targets on the cell surface and intracellularly. The drug combinations selected will be, in part, based on the resistance profiles determined in Aim 1. Upon identifying multitargeted drug combinations that suppress resistance, a final set of studies will be undertaken to analyze TMZ tumoral delivery and the process referred to as vascular normalization, a hallmark of effective drug delivery. As in Aim 1, PK (i.e. drug concentrations) and PD (gene and protein expression) measurements will be obtained in Aim 2 to provide a robust database to formulate PK/PD models for the effective combinations, which is the goal of Aim 3. Specifically, we will build physiologically-based PK/PD models that offer a means to be extrapolated to patients so that PK and PD endpoints can be predicted in brain tumors. The quantitative pharmacological approach underlying the project enables a more seamless pipeline of information to flow into the clinic that hopefully will provide a rational paradigm to design complex multidrug regimens. Recent clinical studies indicate that the effectiveness of antiangiogenic drugs against solid tumors is temporary due to the development of drug resistance. By using preclinical brain tumor models resistant to angiogenesis inhibitors we will develop targeted drug combinations that overcome resistance, and further, enable coadministered cytotoxic drugs to be successfully delivered to the tumor. Quantitative pharmacological models will be derived and extrapolated to patients so that rational and effective multidrug therapy can be implemented in patients

Keywords: 3, 4-dihydro-3-methyl-4-oxoimidazo[5, 1-d]-1, 2, 3, 5-tetrazine-8-carboxamide; 8-carbamoyl-3-methylimidazo[5, 1-d]-1, 2, 3, 5-tetrazin-4(3H)-one; Address; Affect; Alkylating Agents; Alkylators; angiogenesis; Angiogenesis Antagonists; Angiogenesis Blockers; Angiogenesis Inhibitors; Angiogenetic Antagonists; Angiogenic Antagonists; Angiogenic Factor; Angiostatic Agents; Anti-Angiogenetic Agents; Anti-Angiogenic Agents; Anti-Angiogenic Drugs; Antiangiogenesis Agents; antiangiogenic; Antiangiogenic Agents; base; Behavior; blood vessel neoplasm; Blood Vessel Tumor; Blood Vessels; Brain Neoplasia; Brain Neoplasms; Brain Tumors; BZS; c-erbB-1; c-erbB-1 Protein; CD140B; Cell surface; Characteristics; Clinic; clinical data repository; clinical data warehouse; clinical investigation; Clinical Research; Clinical Study; Clinical Trials; Clinical Trials, Unspecified; Complex; cytotoxic; Cytotoxic agent; Cytotoxic drug; Data; Data Banks; Data Bases; data repository; Databank, Electronic; Databanks; Database, Electronic; Databases; design; designing; Development; Drug Combinations; Drug Delivery; Drug Delivery Systems; Drug Interactions; Drug Kinetics; Drug resistance; drug resistant; Drug Targeting; Drug Targetings; drug/agent; Drugs; Effectiveness; EGFR; Ensure; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Kinase; Epidermal Growth Factor Receptor Protein-Tyrosine Kinase; ERBB Protein; erbB-1; erbB-1 Proto-Oncogene Protein; ERBB1; erbBl; Essex brand of temozolomide; Factor, Angiogenesis; Foundations; Gene Expression; Gene Proteins; Generalized Growth; Glial Cell Tumors; Glial Neoplasm; Glial Tumor; Glioma; Goals; Growth; heavy metal lead; heavy metal Pb; HER1; Hybrids; imidazo[5, 1-d]-1, 2, 3, 5-tetrazine-8-carboxamide, 3, 4-dihydro-3-methyl-4-oxo-; improved; in vivo; inhibitor; inhibitor/antagonist; Inhibitors, Angiogenetic; Inhibitors, Angiogenic; Investigation; JTK12; language translation; Lead; Mammals, Mice; Measurement; Measures; Mediating; Medication; methazolastone; MHAM; Mice; MMAC1; Modeling; Molecular; Monitor; Murine; Mus; Nature; Neoplasms in Vascular Tissue; Neoplasms of Neuroglia; Neovascularization Inhibitors; Neuroglial Neoplasm; Neuroglial Tumor; novel; ontogeny; Outcome; Patients; Pb element; PDGF-R-Beta; PDGFR; PDGFR1; PDGFRB; PDGFRB gene; Pharmaceutic Preparations; Pharmaceutical Preparations; pharmacodynamic model; Pharmacodynamics; Pharmacokinetics; Phosphatase and Tensin Homolog; pre-clinical; preclinical; Process; Property; Property, LOINC Axis 2; Prospectuses; prospectuses; Prospectuses [Publication Type]; protein expression; Protein Gene Products; proto-oncogene protein c-erbB-1; PTEN; PTEN gene; PTEN1; public health relevance; Receptor, EGF; Receptor, TGF-alpha; Receptor, Urogastrone; Receptors, Epidermal Growth Factor-Urogastrone; Regimen; relational database; Research; Resistance; resistance mechanism; Resistance profile; resistance to Drug; resistant; resistant mechanism; Resistant profile; resistant to Drug; response; Schering brand of temozolomide; Schering-Plough brand of temozolomide; Series; Solid Neoplasm; Solid Tumor; subcutaneous; Temodal; Temodar; temozolomide; Tissue Growth; Transforming Growth Factor alpha Receptor; Translating; Translatings; Treatment outcome; tumor; Tumor Angiogenesis; tumors in the brain; Tumors of Neuroglia; vascular; Vascular Endothelial Cell Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor; Vascular Neoplasms; Vascular Permeability Factor Receptor; Vascular Tissue Tumor; Vascular Tumor; VEGF Receptors; VEGFR; VPF Receptor

Relevance: Interactions Between Cytotoxic and Antiangiogenic Drugs Recent clinical studies indicate that the effectiveness of antiangiogenic drugs against solid tumors is temporary due to the development of drug resistance. By using preclinical brain tumor models resistant to angiogenesis inhibitors we will develop targeted drug combinations that overcome resistance, and further, enable coadministered cytotoxic drugs to be successfully delivered to the tumor. Quantitative pharmacological models will be derived and extrapolated to patients so that rational and effective multidrug therapy can be implemented in patients

Project start date: 1998-02-01

Project end date: 2015-11-30

Budget start date: 10-DEC-2010

Budget end date: 30-NOV-2011

PFA/PA: PA-10-067

2R01CA072937-09 (2011): $217570

COOKSTOVE REPLACEMENT FOR PREVENTION OF ARI AND LOW BIRTHWEIGHT IN NEPAL

M James, Professor
Johns Hopkins Universitycity: Baltimore    country: United States (us)

Grant 5R01ES015558-04 from National Institute Of Environmental Health Sciences

Abstract: The goals of our research program are to determine which interventions are most effective at reducing the burden of mortality and morbidity among women and children in high-risk populations in developing countries. At our field site in southern Nepal, acute respiratory illness (ARI) has been a leading cause of mortality among young children. Besides immunization there is little evidence for effective primary preventive approaches for ARI on a population basis. Low birth weight is highly prevalent in this population as well affecting approximately 30% of live born infants. Low birth weight is a key determinant of neonatal mortality and has also been resistant to cost-effective interventions in resource poor settings. Given the lack of appropriate interventions for poor, rural areas in developing countries and the strong observational association between open burning of biomass fuel sources and ARI in young children and low birth weight, we have designed a community-based randomized trial to determine if reductions in household indoor smoke exposure can reduce the incidence and duration of acute lower respiratory infections in children <36 months of age and low birthweight among newborn infants. Household indoor smoke reduction will be accomplished by replacing the current cook stove in the household with a locally appropriate, inexpensive model that is more efficient and vented to the exterior. In addition, we will assess the impact on respiratory function and symptoms among adults in the household, including women of reproductive age, and compare total fuel consumption and time spent collecting fuel for the household. The project is a cluster-randomized, community-based trial of cookstove replacement in a rural population of southern Nepal. Households will be randomized to receive replacement of their cook stove with an appropriately designed, efficient stove that is vented to the exterior at different time periods during the course of the study. An initial period of surveillance for ARI and low birth weight will establish a baseline rate for all clusters. This will be followed by the randomized, serial replacements of cook stoves over a 12 month period. Surveillance will continue throughout this period and for an additional 6 -18 months depending on when the stove was replaced. Two cohorts of sectors will enter the trial sequentially. Measurement of indoor air particulate concentration will be conducted in a sample of households before and after stove replacement. The analysis will focus on estimating the impact on incidence of ARI in children and low birth weight among live births as a result of stove replacement. We will also assess the relative efficiency of the new stoves by identifying fuel consumption in the household and time spent collecting fuel. Approximately 4200 children 1-35 months of age will be required to observe a minimum 10% reduction in risk of ARI with 90% power. Given the expected number of live births to occur in these clusters, we can detect a 50 gram difference in birthweight with over 90% power and a type I error of 5%. Lower respiratory infections and low birthweight are leading causes of death among young children in developing countries. There are few preventative measures for these problems that are cost-effective in the context of severe resource constraints. We propose to test an inexpensive approach to reducing the risk of these infections and low birthweight by reducing the exposure to airborne fine particles and carbon monoxide caused by the open burning of biomass fuel sources in the home by replacing the cookstoves with an improved, more efficient, and vented stove. If the replacement of these stoves works to reduce respiratory infections and low birthweight, significant number of children´s lives can be saved at very low cost

Keywords: Acute; Address; Adult; Affect; Age; Age-Months; Air; Antibiotic Therapy; Area; Asthma; base; Biomass; Blood Pressure; Burn injury; Carbon Monoxide; Cause of Death; Child; Child Mortality; Chlorhexidine; Chronic Disease; cohort; Communicable Diseases; Communities; Consumption; cooking; cost; cost effective; cost effectiveness; design; Developing Countries; effective intervention; effective therapy; Effectiveness of Interventions; Environmental Exposure; Evaluation; Exposure to; Folate; Funding; Goals; Growth; Health; high risk; Home environment; Household; Immunization; improved; Incidence; Infant; Infection; interest; Intervention; Iron; Life; Live Birth; Low Birth Weight Infant; maternal cigarette smoking; Measurement; Measures; member; micronutrient deficiency; Modeling; Morbidity - disease rate; Mortality Vital Statistics; National Institute of Environmental Health Sciences; neonatal morbidity; Neonatal Mortality; Nepal; neurodevelopment; Newborn Infant; Outcome; particle; Particulate; Personal Satisfaction; Policy Maker; Population; population based; Pregnancy; Pregnant Women; Prevention; Preventive; programs; pulmonary function; Randomized; randomized trial; Relative (related person); reproductive; Research; Research Personnel; Resistance; Resources; respiratory; Respiratory physiology; Respiratory Tract Infections; Risk; rural area; Rural Population; Sampling; Severities; Site; Skin; Smoke; Source; Spirometry; Supplementation; Symptoms; Testing; therapy design; Time; Umbilical cord structure; Vent; Woman; Work; Zinc

Project start date: 2008-09-17

Project end date: 2013-05-31

Budget start date: 1-JUN-2011

Budget end date: 31-MAY-2012

PFA/PA: PA-07-070

5R01ES015558-04 (2011): $861658

GENE THERAPY FOR UREA CYCLE DISORDERS

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Grant 5P01HD057247-04 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development

Abstract: The goal of this application is to develop an effective gene therapy for ornithine transcarbamylase (OTC) deficiency. Studies begin with the development of an optimal AAV vector called the "clinical candidate" in Project I. Additional studies will be performed in this project to evaluate potential immunologic barriers to successful liver directed gene therapy. The clinical candidate will be evaluated for safety and efficacy in OTC deficient mice and nonhuman primates in Project II. The molecular state of the vector genome and its potential oncogenicity will be studied in Project III. Cores for vector production, cell morphology, animal models, and administration will provide support to the projects. An External Scientific Advisory Committee and an Ethics Advisory Board will be convened to provide advice to the program leadership. At the completion of this four year program, the investigators hope to provide an AAV vector with preclinical data to support its translation into humans

Project start date: 2008-04-01

Project end date: 2012-03-31

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5P01HD057247-04 (2011): $1122053

UNDERSTANDING COUPLED TRANSFERS OF ELECTRONS AND PROTONS RELEVANT TO BIOLOGICAL C

M James, Professor
University Of Washingtoncity: Seattle    country: United States (us)

Grant 5R01GM050422-16 from National Institute Of General Medical Sciences

Abstract: The goal of the proposed research is to develop a fundamental and predictive understanding of processes in which electron transfer and proton transfer are coupled. Such proton-coupled electron transfer (PCET) processes are critical in many areas of biology, from bioenergetics to the catalytic cycles of numerous metalloenzymes, to the trapping of reactive oxygen species. The studies proposed here focus on reactions in which one proton and one electron transfer in a single kinetic step, termed concerted proton and electron transfer (CPET). One set of projects are designed to build a unified picture of hydrogen atom transfer (HAT) reactions. These are processes which involve the concerted transfer of an electron and a proton from a single donor to a single acceptor. Model systems are being developed to develop the general principles of HAT and to understand specifically the reactivity of iron-histidine cofactors, including qui oxidation by the Rieske iron-sulfur cluster in the mitochondrial bc1 complex and the chemistry of bis(histidine)-ligated hemes. A second set of projects examine

Relevance: separated CPET´ processes, in which the electron and proton transfer to different acceptors. Such reactions have received little attention but are likely to be involved in a wide range of biological redox reactions. The best-studied example is the formation of the tyrosyl-Z radical in photosystem II; models for this process are proposed. In each of these areas, some systems model specific biochemical reactions while others are designed to probe general principles. For example, the proposed studies of oxidation of phenol-base compounds will probe how framework motions affect hydrogen transfers, an issue of much current debate regarding the origins of enzymatic catalysis. These studies will provide connections to current theoretical treatments of CPET, building on our demonstration that a range of hydrogen atom transfer reactions follow Marcus theory. Brought together, these studies will develop the principles of coupled proton-electron transfers and produce new insights into a wide range of biological processes. Such fundamental principles of chemical processes that occur in biology are part of the foundation for biomedical advances. For instance, the detailed knowledge available for electron transfer reactions has proven to be of great importance in biology. The work proposed aims to build a similarly valuable understanding for reactions that involve coupled transfers of electrons and protons. This project is developing a predictive understanding of a class of chemical processes that occur widely in biology but have received little study: coupled transfers of electrons and protons. These chemical reactions are central to fields as diverse as bioenergetics and the action of antioxidants. A fundamental understanding of these chemical processes is a part of the foundation on which biomedical advances will be built

Project start date: 1995-02-01

Project end date: 2012-08-31

Budget start date: 1-SEP-2011

Budget end date: 31-AUG-2012

PFA/PA: SUPPORT FOR GENESIS

M James
University Of Texas Hlth Sci Ctr San Antcity: San Antonio    country: United States (us)

Grant 5R01NS049288-08 from National Institute Of Neurological Disorders And Stroke

Abstract: The General Neural Simulation System (GENESIS) was first released for general use in 1988 as part of the first Methods in Computational Neuroscience Meeting at the Marine Biological Laboratory in Woods Hole, MA. Since its original release 19 years ago, GENESIS has provided one of the foundations for the growth of computational neuroscience both continuing to support education as well as research. With respect to summer courses, GENESIS continues to be part of the course in Woods Hole, as well as courses offered in the European Union, Mexico, India, Japan, and recently a new course in Brazil for Latin America. At last count GENESIS has also provided support for courses in at least 61 colleges and universities around the world where it has been used both as an instruction tool in realistic modeling of the nervous system, and as a simulation based tool for neurobiological education in general. The Book of GENESIS (Bower and Beeman, 1994, 1998), which was designed to support both computational and neurobiological instruction has sold more than 6,000 copies worldwide with more than double that number of copies having been distributed with the text now freely available on the Internet (Bower and Beeman, 2003). This substantial support for the use of GENESIS in instruction has also provided the base for extensive and growing use of this software system in biological research. Although certainly an underestimate, we are aware of 297 published GENESIS-dependent peer reviewed journal articles and book chapters not directly related to research in the P.I.´s laboratory. Further, more than a third of those reports (103) have been published during the period January 2004 to June 2007 (approximately the duration of the current NIH grant). As measured by the subjects of these publications, use of GENESIS has a growing emphasis on two types of simulations, those of large scale networks (using the parallel version of the platform), and those involving molecular biological modeling. GENESIS remains the only simulation system specifically designed from the outset to link multi-scale modeling efforts. With this grant, we seek continuing funding to support the expanding use of GENESIS, but also and importantly, to complete the implementation of a new version of the GENESIS platform. While maintaining backwards compatibility, GENESIS 3.0 is a complete rewrite and redesign of the system, replacing a 19-year-old structure. The rewrite will update and upgrade core GENESIS functionality while adding important new features, including model history control and tracking, a new web browser interface, and new features to support education as well as scholarly publication of models. Biological simulation technology is becoming an increasingly important part of both basic and translational research in medicine. The GENESIS system supported by this grant is one of the most used software systems supporting this type of research in the US, and will provide continuing support for the use and further development of this modeling platform. Newly proposed extensions include applications to medical education and scientific publishing as well as the continuing efforts to understand brain function and dysfunction

Keywords: 19 year old; Access to Information; base; Basic Science; Biological; Biological Models; biological research; Book Chapters; Books; Brain; Brazil; clinical Diagnosis; Code; Collaborations; college; Communities; computational neuroscience; Computer software; Continuing Education; Data; Databases; design; Development; Educational aspects; Educational Curriculum; Educational process of instructing; European Union; Extensible Markup Language; flexibility; Foundations; Functional disorder; Funding; Grant; Growth; India; Instruction; Internet; Japan; journal article; Laboratories; Latin America; Link; Maintenance; Marines; Measures; Medical; Medical Education; medical schools; Medical Students; Medicine; meetings; Methods; Mexico; model development; Modeling; Molecular; Morphology; multi-scale modeling; Nature; Nervous system structure; Neurobiology; Neurons; Online Systems; open source; Pathway interactions; Peer Review; Physicians; Play; prototype; Publications; Publishing; Recording of previous events; relating to nervous system; Reporting; Research; Role; Series; simulation; software development; software systems; Stream; Structure; Students; Support System; System; Technology; Text; Time; tool; Translational Research; United States National Institutes of Health; Universities; Update; Wood material

Project start date: 2004-09-01

Project end date: 2013-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PAR-07-235

5R01NS049288-08 (2011): $286513

STRUCTURE/FUNCTION STUDIES OF DNA REPLICATION INITIATION

M James, Professor
University Of California Berkeleycity: Berkeley    country: United States (us)

Grant 5R01GM071747-07 from National Institute Of General Medical Sciences

Abstract: The faithful propagation of genetic material is essential to life. A central prerequisite to the copying of DNA is the dedicated assembly of replicative machineries at proper sites on the chromosome. This replication "initiation" event depends on multiple components, including 1) initiators, ATPases that bind origins and recruit other proteins to the replication start site, 2) helicase-loaders, which deposit replicative helicases onto DNA, and 3) primases, enzymes that create short oligonucleotides for extension by DNA polymerases. A long-term objective of our research has been to understand the molecular structure/function relationships that govern the initiation of DNA replication. Many of the proteins responsible for initiation have been identified, and a preliminary framework for their action is in place. However, there remains a host of outstanding questions regarding how these factors act individually and cooperatively to construct a competent replication fork. Our prior efforts in this area have generated new mechanistic models for initiator action, helicase loading, and priming that we are now in an ideal position to test. Using a combination of structural, biochemical, and biophysical methods we aim to 1) Define how ATP controls DNA binding and remodeling by prokaryotic initiators, 2) Determine how ATP regulates bacterial helicase-loader assembly and function, and 3) Establish how the bacterial primase synthesizes primers and is inhibited by disparate types of small-molecule agents, including the stringent response regulator, (p)ppGpp. Together, these efforts will impact a number of scientific fronts, from understanding how origins are engaged and restructured to promote replisome construction, to defining how chemical energy and small-molecule inhibitors control the function of dynamic macromolecular machines. Knowledge of these processes is necessary for understanding how cells ultimately avoid initiation errors linked to genomic instabilities, transformation, and neoplasia. The faithful propagation of genetic material is essential to life. The goal of our research is understand the structure/function relationships that govern the initiation of DNA replication at a molecular level. Knowledge of these events is necessary for understanding how cells ultimately avoid errors linked to genomic instabilities and cancer onset

Keywords: Area; ATP phosphohydrolase; ATPase Domain; Award; base; Binding (Molecular Function); Biochemical; Biological Assay; Biological Factors; cell transformation; Cells; Chemicals; Chromosomes; Complex; crosslink; Deposition; DNA; DNA Binding; DNA Primase; DNA replication origin; DNA-Directed DNA Polymerase; DnaB helicase; DnaG Protein; drug development; ds-DNA; Enzymes; Escherichia coli; Event; Family; Foundations; Genetic Materials; Genomic Instability; Goals; Hand; Health; helicase; Helix-Turn-Helix Motifs; Homo; Image; inhibitor/antagonist; Investigation; Knowledge; Lead; Life; Link; Malignant Neoplasms; melting; Methods; Modeling; Molecular; Molecular Structure; Mutagenesis; Neoplasms; novel; Nucleic Acids; Nucleotides; Oligonucleotides; paralogous gene; Peptide Initiation Factors; Polymerase; Positioning Attribute; Process; Prokaryotic Cells; Proteins; Recruitment Activity; Replication Initiation; Replication Origin; Research; response; scaffold; Single-Stranded DNA; Site; small molecule; Structure; Structure-Activity Relationship; Testing; Work

Relevance: The faithful propagation of genetic material is essential to life. The goal of our research is understand the structure/function relationships that govern the initiation of DNA replication at a molecular level. Knowledge of these events is necessary for understanding how cells ultimately avoid errors linked to genomic instabilities and cancer onset

Project start date: 2005-05-01

Project end date: 2013-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-07-070

5R01GM071747-07 (2011): $293836

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NIGROSTRIATAL DOPAMINE FUNCTION

M James, Professor
Rutgers The State Univ Of Nj Newarkcity: Newark    country: United States (us)

Grant 5R01NS034865-13 from National Institute Of Neurological Disorders And Stroke

Abstract: The basal ganglia is an essential component of the central circuitry controlling voluntary movement as well as sensorimotor integration, motor and non-motor learning, and a number of higher cognitive functions. The major input structure of the basal ganglia is the striatum, comprised mostly of medium sized GABAergic spiny projection neurons (MSNs) that make up about 95% of striatal neurons in the rodent. The remaining neurons consist of cholinergic interneurons and 3 types of GABAergic interneurons. The GABAergic interneurons play a crucial role in striatal function by participating in a powerful feedforward inhibitory circuit that affects spike timing in the spiny neurons. Dopamine (DA), originating in the substantia nigra, has long been recognized to play an essential role in striatal function, and it is the degeneration of the nigrostriatal DAergic pathway that is the cause of Parkinson´s disease, a progressive and incurable disorder that affects between 1 and 1.5 million Americans. In addition to the cell types listed above, a population of striatal neurons has been recognized that expresses tyrosine hydroxylase (TH), the rate-limiting enzyme in the synthesis of DA. In primates essentially all of these neurons also express the DA transporter (DAT) suggesting strongly that they are DAergic. These neurons also express glutamate decarboxylase, the enzyme responsible for the synthesis of GABA and a common marker for GABAergic neurons. The numbers of these neurons increases several-fold following DA denervation, and some of these neurons have been shown to express L-amino acid decarboxylase (AADC) and the vesicular monoamine transporter (VMAT). These neurons could represent a heretofore-unappreciated source of striatal DA and a potentially useful source of compensation for DA loss in idiopathic Parkinson´s disease as well as a potential target for novel therapeutic approaches to the treatment of the disease. These striatal TH+ neurons represent novel components of the intrastriatal circuitry about which little or nothing is known, since they have never been recorded from, but only studied with immunocytochemistry. We used striatal slices from mice genetically engineered to express enhanced green fluorescent protein (EGFP) under the control of the TH promoter to obtain visually guided recording from these neurons in brain slices. We have identified 4 electrophysiologically distinct subtypes and provide preliminary data on their efferent and afferent synaptic connectivity. Using these mice, both untreated and after unilateral dopaminergic denervation and/or L-DOPA replacement therapy, we will describe the basic electrophysiological properties of striatal DA neurons, their afferent and efferent connectivity, compensatory changes in DA depletion animal models of PD, and their role in striatal DA and GABAergic neurotransmission. In addition, these mice afford a novel way to study the electrophysiological and anatomical properties of uncharacterized populations of striatal interneurons that have been difficult or impossible to study previously in any systematic way. PUBLIC HEALTH RELEVANCE Parkinson´s disease (PD) is the most common neurological disorder, affecting nearly 15% of people over the age of 65 and over 50% of people over the age of 85. This translates to approximately 1.5 million Americans. The disease is progressive and incurable. PD is caused by a loss of dopamine input to the neostriatum, a brain structure that controls voluntary movement. In this project we will characterize electrophysiologically, neurochemically and neuroanatomically novel dopamine-like neurons in the striatum that have the potential to serve as the focal point for novel therapeutic approaches to ameliorating the symptoms of the PD

Keywords: Action Potentials; Adult; Affect; Age; Agonist; American; Amino Acids; Anatomy; Animal Model; Animals; Basal Ganglia; Bathing; biocytin; Brain; Brightfield Microscopy; Calcium-Binding Proteins; Carboxy-Lyases; Cell Count; cell type; Cells; cholinergic; cognitive function; Contralateral; Corpus striatum structure; Coupled; Coupling; Data; Denervation; Development; Disease; Dopamine; Dopamine Receptor; dopamine transporter; dopaminergic neuron; Elderly; Engineering; enhanced green fluorescent protein; Enzymes; extracellular; Financial compensation; Fluorescence; gamma-Aminobutyric Acid; Genetically Engineered Mouse; Globus Pallidus; Glutamate Decarboxylase; Health; immunocytochemistry; In Vitro; Incidence; Injection of therapeutic agent; Interneurons; Ipsilateral; Label; Learning; Lesion; Lighting; Measures; median forebrain bundle; Mediating; Membrane Potentials; Minor; Molecular Profiling; Morphology; motor learning; Movement; Mus; Neostriatum; nervous system disorder; neurochemistry; Neurons; neurotransmission; nigrostriatal pathway; Nomarski Interference Contrast Microscopy; novel; novel therapeutic intervention; Oxidopamine; Parkinson Disease; Pathway interactions; Pharmacology; Phenotype; Physiologic pulse; Physiology; Play; Population; postsynaptic; presynaptic; Primates; Probability; Promotor (Genetics); Property; Replacement Therapy; research study; Resistance; response; retrograde transport; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Role; Slice; Source; Structure; Substantia nigra structure; Symptoms; Synapses; synthetic enzyme; Testing; Time; Training; Transgenic Mice; Translating; Tyrosine 3-Monooxygenase; vesicular monoamine transporter; voltage; voltage clamp; Voltage-Clamp Technics; Whole-Cell Recordings

Relevance: Parkinson´s disease (PD) is the most common neurological disorder, affecting nearly 15% of people over the age of 65 and over 50% of people over the age of 85. This translates to approximately 1.5 million Americans. The disease is progressive and incurable. PD is caused by a loss of dopamine input to the neostriatum, a brain structure that controls voluntary movement. In this project we will characterize electrophysiologically, neurochemically and neuroanatomically novel dopamine-like neurons in the striatum that have the potential to serve as the focal point for novel therapeutic approaches to ameliorating the symptoms of the PD

Project start date: 1997-02-01

Project end date: 2013-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PA-07-070

5R01NS034865-13 (2011): $331209

PATHOGENESIS OF HIV-RELATED EMPHYSEMA: T CELLS, SMOKING*

M James
University Of Michigan At Ann Arborcity: Ann Arbor    country: United States (us)

Grant 5R01HL083482-05 from National Heart, Lung, And Blood Institute

Abstract: The advent of highly active anti-retroviral therapy (HAART) has permitted significant increases in survival and in quality of life in those HIV-infected individuals with access to advanced medical care and pharmaceuticals. With increased survival, and with effective prophylaxis and treatment for many previously lethal opportunistic infections, the manifestations of chronic diseases have become more prevalent in HIV-infected individuals. One such manifestation of chronic HIV infection is emphysema. Recent literature clearly documents the existence of accelerated emphysema, particularly in HIV-infected individuals who smoke cigarettes. The clinical literature has suggested that the CD8+ T cell may be important in the in the pathogenesis of HIV-related emphysema, but this relationship remains associational and speculative. Pneumocystis pneumonia remains an important cause of morbidity and mortality in HIV-infected individuals. Recently, the existence of carrier states for this infection have been identified, both in HIV-infected individuals and in those individuals who provide their care. Furthermore, Pneumocystis carriage is common in smokers with chronic obstructive pulmonary disease. Whether repeated bouts of Pneumocystis colonization or carriage contribute to lung damage, particularly in interaction with cigarette smoke, has not been investigated experimentally. Although HIV infection produces wide-ranging immunologic effects, depletion of CD4+ T cells remains the immunologic hallmark of HIV infection. Our laboratories have many years of experience studying the lungs of chronically CD4-depleted mice, as an immunologically valid animal model of HIV-induced immunosuppression. Furthermore, we have characterized the immunologic responses to chronic Pneumocystis infection in these CD4-depleted mice, a well-accepted animal model of this important opportunistic infection. In these investigations, the CD8+ T cell has emerged as an important mediator of lung inflammation and lung damage. Most recently, we have acquired expertise in the development and analysis of cigarette smoke-induced emphysema in mice. We hypothesize that, during persistent depletion of CD4+ T cells, chronic Pneumocystis colonization and cigarette smoke interact to cause emphysema. The development of emphysema is mediated by CD8+ T cells in the lung, and results from cytokine and perforin release. Furthermore, we hypothesize that CD8+ T cells drive the development of emphysema by damage to alveolar epithelial cells. RELEVANCE Chronic manifestations of HIV infection, including emphysema, are likely to present increasing clinical challenges in the era of HAART, but the pathogenesis of HIV-related emphysema is not understood. Using immunologically relevant animal models, this project will investigate the pathogenesis of smoking-related emphysema during chronic immunosuppression, and will examine Pneumocystis as a relevant cofactor in the development of emphysema

Keywords: Acquired Immune Deficiency Syndrome Virus; Acquired Immunodeficiency Syndrome Virus; AIDS Virus; Alveolar; Alveolar Macrophages; Alveolitis; Alveolus; Animal Model; Animal Models and Related Studies; anti-retroviral therapy, highly active; Antiretroviral Therapy, Highly Active; Bronchial Alveolus; Caring; Carrier State; CD4 lymphocyte; CD4 Positive T Lymphocytes; CD4 T cells; CD4+ T cell; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; CD8; CD8B; CD8B1; CD8B1 gene; cell damage; cell injury; Cells; Cells, CD4; Cellular injury; Chronic; Chronic Disease; chronic disease/disorder; chronic disorder; Chronic Illness; Chronic Obstructive Airway Disease; Chronic Obstructive Lung Disease; cigarette smoke; Cigarette smoke-induced emphysema; cigarette smoking; Clinical; COAD; cofactor; COPD; cytokine; cytolysin; Dendritic Cells; Development; disease carrier state; Emphysema; Epithelial Cells; experience; HAART; helper T cell; Highly Active Antiretroviral Therapy; HIV; HIV Infections; HTLV-III; HTLV-III Infections; HTLV-III-LAV Infections; Human; Human Immunodeficiency Viruses; Human T-Cell Leukemia Virus Type III; Human T-Cell Lymphotropic Virus Type III; Human T-Lymphotropic Virus Type III; Human, General; Immunocompromised; Immunocompromised Host; Immunocompromised Patient; Immunologic, Immunochemical; Immunologics; Immunosuppressed Host; immunosuppressed patient; immunosuppression; Immunosuppression Effect; Immunosuppressions (Physiology); Immunosuppressive Effect; In Vitro; Individual; Infection; Investigation; Investigators; Laboratories; latent infection; LAV-HTLV-III; Literature; living system; longitudinal human study; Lung; Lung diseases; lung disorder; Lung Inflammation; Lymphadenopathy-Associated Virus; lymphocyte pore-forming protein; LYT3; Macrophages, Alveolar; Mammals, Mice; Man (Taxonomy); Man, Modern; Mediating; Mediator; Mediator of Activation; Mediator of activation protein; Medical; Methods and Techniques; Methods, Other; Mice; model organism; Modeling; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Murine; Mus; Natural immunosuppression; Opportunistic Infections; Organism; Pathogenesis; perforin; Pharmaceutical Cares; Pneumocystis; Pneumocystis carinii Infections; Pneumocystis carinii Pneumonia; Pneumocystis Infections; Pneumocystosis; Pneumonia; Pneumonia, Interstitial Plasma Cell; Pneumonia, Pneumocystis; Pneumonia, Pneumocystis carinii; Pneumonitis; Position; Positioning Attribute; Prevention Measures; programs; Programs (PT); Programs [Publication Type]; Prophylactic treatment; Prophylaxis; pulmonary; Pulmonary Disease, Chronic Obstructive; Pulmonary Diseases; Pulmonary Disorder; Pulmonary Emphysema; Pulmonary Inflammation; Pulmonary Macrophages; QOL; Quality of life; Research Personnel; Researchers; respiratory; Respiratory Disease; Respiratory Disorder; Respiratory System Disease; Respiratory System Disorder; Respiratory System, Lung; response; Role; Smoke; smoke cigarette; smoke of cigarettes; Smoker; Smoking; smoking-induced emphysema; social role; T-Cells; T-Lymphocyte; T-Lymphotropic Virus Type III Infections, Human; T4 Cells; T4 Lymphocytes; Techniques; Testing; thymus derived lymphocyte; Thymus-Dependent Lymphocytes; Veiled Cells; Virus-HIV

Project start date: 2005-09-29

Project end date: 2011-06-30

Budget start date: 1-JUL-2009

Budget end date: 30-JUN-2011

PFA/PA: RFA-HL-04-031

5R01HL083482-05 (2009): $325585

ADMINISTRATIVE CORE

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Abstract: The administrative home for the management of this Center and the various participating cores is the Gene Therapy Program of the Division of Transfusion Medicine of the Department of Pathology and Lab Animal Medicine which is headed by the PI of this grant, Dr. James M. Wilson. The Dean of the Medical School provides financial support to this program. Dr. Maria Limberis the Gene Therapy Department is the Co-Pi of the grant and Ms. Kelly D. Reynolds, who is the Director of Finance and Operations for the Gene Therapy Program, provides all administrative support for the activities of the Center. A number of administrative vehicles are in place to assure the Center and its participating cores meet the objectives of the program. Independent weekly core management meetings are held to review activities of the cores including the following Vector and Immunology Core Management meeting, Cell Morphology Management meeting, and Animal Models Core Management meeting. Internal and External Oversight Committees are in place to review the activities of the Center. The Gene Therapy Program coordinates a Gene Therapy Seminar Series which is partially sponsored by this Center. Finally, the Administrative Core oversees all aspects of the Pilot and Feasibility Program

Keywords: Advisory Committees; Animal Model; Animals; Cellular Morphology; Ethics; Financial Support; gene therapy; Grant; Head; Hereditary Disease; Home environment; Immunology; medical schools; Medicine; meetings; Molecular; operation; Pathology; programs; Series; Transfusion; vector

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5P30DK047757-18_7579 (2011): $270119

SUBPOPULATION DIFFERENCES IN INTERVENTION EFFICACY FOR COLLEGE DRINKERS

M James, Assistant Professor
Old Dominion Universitycity: Norfolk    country: United States (us)

Grant 5R03AA018771-02 from National Institute On Alcohol Abuse And Alcoholism

Abstract: Subpopulation Differences in Intervention Efficacy for College Drinkers. Despite prevention efforts, college binge drinking continues to account for a myriad of negative personal and social alcohol-related consequences. Brief motivational interventions (BMIs) have proved efficacious in reducing college alcohol consumption; however, averaging across people, reported intervention effects tend to be small and short-lived. It is often of more interest to understand who responds to an intervention rather than determine the average effect across different subpopulations. Instead of characterizing intervention efficacy for a "typical" college drinker, the purpose of this research is to characterize intervention effects across different subpopulations, or types, of drinkers. Moderation analyses test hypothesized factors that may influence intervention efficacy. In contrast to traditional moderation analyses, growth mixture modeling (GMM) empirically explores the data to identify homogenous longitudinal patterns in alcohol consumption without need of pre-identified predictors. Using GMMs, past research has identified up to five types of drinkers regarding naturally-occurring trajectories of alcohol consumption over time; however, the proposed project takes a novel approach by exploring drinking patterns following intervention. The proposed project includes secondary data analysis of 3 randomized clinical trials with a combined total of 1,384 college-student drinkers (509 volunteers, 875 mandated as a result of alcohol violations), whose drinking behaviors were assessed at baseline, and at 1, 6, and 12 months post-intervention. Participants were randomized into a BMI (n = 602), 1 of 2 computerized interventions (Alcohol 101, n = 271; Alcohol EDU, n = 167), or a no-intervention control condition (n = 344). The four specific aims are a) to identify how many different subpopulations of college drinkers exist regarding short- and long-term response to an alcohol BMI; b) to characterize BMI efficacy relative to computerized intervention and a control for each type of drinker; c) to use demographic and psychological explanatory variables to characterize each drinker subpopulation; and d) ascertain how drinker subpopulations relate to high-risk alcohol consequences. Discontinuous GMMs will be used to ascertain how different types of drinkers uniquely respond following intervention from baseline to 1-month (short-term effect) and from 1-month to 12- months (long-term effect), as well as to identify relevant psychological predictors of drinker subpopulations among both sanctioned individuals and volunteers. Results of this research can be used to understand who is most receptive to BMI intervention, to guide screening efforts to reliably detect high-risk individuals, and to use the information about the most resistant subpopulation(s) to inform future intervention development. Subpopulation Differences in Intervention Efficacy for College Drinkers The proposed research will use innovative and advanced statistical modeling to identify subpopulation characteristics of individuals who respond and do not respond to interventions designed to reduce alcohol consumption among college drinkers. The results of this research can be used to understand who is most receptive to brief motivational intervention, to guide screening efforts to reliably detect high-risk individuals, and to use the information about the most resistant subpopulation(s) to inform future intervention development. Improved screening and intervention methods are critical to reducing personal and social alcohol-related consequences among college populations

Keywords: Accounting; Affect; alcohol consequences; Alcohol consumption; alcohol intervention; Alcohol or Other Drugs use; alcohol related consequences; alcohol response; alcohol violation; Alcoholic beverage heavy drinker; Alcohols; assault; binge drinking; brief motivational intervention; Cessation of life; Characteristics; college; college drinker; computerized; Data; Data Analyses; Data Collection; demographics; drinking; drinking behavior; Evaluation; Future; Gender; Growth; Heterogeneity; high risk; improved; Individual; Informal Social Control; Injury; innovation; interest; Intervention; intervention effect; Intervention Studies; Life; Light; Long-Term Effects; Methods; Modeling; Nature; neglect; novel strategies; Participant; Pattern; Population; post intervention; Prevention; psychologic; public health relevance; Randomized; Randomized Clinical Trials; Readiness; recreational drug use; Relative (related person); Reporting; Research; Research Personnel; Resistance; Sampling; Schools; Screening procedure; sexual assault; social; Statistical Models; statistics; Students; Testing; therapy design; therapy development; Time; Treatment Efficacy; Typology; university student; volunteer

Relevance: Subpopulation Differences in Intervention Efficacy for College Drinkers The proposed research will use innovative and advanced statistical modeling to identify subpopulation characteristics of individuals who respond and do not respond to interventions designed to reduce alcohol consumption among college drinkers. The results of this research can be used to understand who is most receptive to brief motivational intervention, to guide screening efforts to reliably detect high-risk individuals, and to use the information about the most resistant subpopulation(s) to inform future intervention development. Improved screening and intervention methods are critical to reducing personal and social alcohol-related consequences among college populations

Project start date: 2010-07-01

Project end date: 2012-03-31

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

PFA/PA: PA-07-408

5R03AA018771-02 (2011): $70121

EXPRESSION OF SIMIAM VIRUS 40 T ANTIGENS IN INTESTINE

M James, Professor
University Of Pittsburgh At Pittsburghcity: Pittsburgh    country: United States (us)

Grant 5R01CA098956-08 from National Cancer Institute

Abstract: Simian virus 40 (SV40) encodes a powerful oncoprotein, large T antigen, that is capable of inducing tumors in animals and transforming cells in culture. T antigen acts in part by inhibiting the action of two tumor suppressor proteins, members of the retinoblastoma (Rb) family, and p53. Several lines of evidence indicate that T antigen has additional functions that contribute to transformation as well. Most studies of SV40 transformation have made use of cultured cells. In this application we propose to study the effects of T antigen expression on the growth-arrested enterocytes and continuously cycling progenitor cells of the mouse small intestine. Because the cycling cells residing in the crypts and the terminally differentiated cells occupying the villi can be isolated, this system allows a combined genetic and biochemical approach to dissecting T antigen´s transforming functions. We have found that the transformation of terminally- differentiated enterocytes is dependent on the transcription factor E2F2. In contrast normal intestinal crypt cell proliferation does not require any of the activating E2Fs. The goals of this application are to (1) determine if T antigen action on the Rb and p53 pathways is sufficient for transformation of crypt epithelial cells; (2) assess the mechanisms allowing cell proliferation to proceed in normal crypts in the absence of E2F1-2-3 and, (3) determine whether the activating E2Fs (E2F1, E2F2, and E2F3a) are required to establish repression of E2F-target genes? This project uses the DNA tumor virus SV40 to probe mechanisms of cellular growth control and tissue homeostasis. Understanding these mechanisms should lead to better therapies for cancer and may suggest approaches for the treatment of certain degenerative diseases

Keywords: Ablation; Address; Amino Acids; Animals; Back; base; Binding (Molecular Function); Biochemical; cancer therapy; Cell Culture Techniques; Cell Cycle; cell growth; Cell Proliferation; cell transformation; cell type; Cells; chromatin modification; Complex; crypt cell; Cultured Cells; Data; Defect; Degenerative Disorder; DNA Damage; DNA Tumor Viruses; Dysplasia; E2F1 gene; E2F2 transcription factor; Enterocytes; Epithelial Cells; Epithelium; Exhibits; Family; Fractionation; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Goals; Growth; Health; Homeostasis; Hyperplasia; intestinal crypt; Intestines; Knock-out; Knockout Mice; Lead; Length; Mediating; member; Messenger RNA; Modeling; Molecular; Mus; mutant; Neoplasms; Normal Cell; Null Lymphocytes; Oncogene Proteins; Pathway interactions; Play; Process; Proliferating; Proteins; Repression; research study; response; Retinoblastoma; Retinoblastoma Protein; retinoblastoma tumor suppressor; Role; sialosyl-T antigen; Simian virus 40; Small Intestines; stem; Stem cells; System; Testing; theories; Tissues; TP53 gene; Transcriptional Activation; Transgenic Mice; tumor; Tumor Antigens; Tumor Suppressor Proteins; tumorigenesis; Variant; Villus; Viral Tumor Antigens; Virus; Work

Project start date: 2002-12-01

Project end date: 2014-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-07-070

5R01CA098956-08 (2011): $290409

REGULATION OF THE SALMONELLA PATHOGENICITY ISLAND 1 TYPE III SECRETION SYSTEM

M James, Associate Professor
University Of Illinois Urbana-champaigncity: Champaign    country: United States (us)

Grant 5R01AI080705-02 from National Institute Of Allergy And Infectious Diseases

Abstract: Salmonella cause 1.4 million cases of gastroenteritis and enteric fever per year in the US and lead all other foodborne bacterial pathogens as a cause of death. A prerequisite for Salmonella to cause both intestinal and systemic disease is the direct injection of effector proteins into host intestinal epithelial cells via a Type Three Secretion System (T3SS) encoded on Salmonella Pathogenicity Island 1 (SPI1). These effector proteins induce inflammatory diarrhea and bacterial invasion. Expression of the SPI1 T3SS is tightly regulated in response to environmental signals from a variety of global regulatory systems. Our long term goal is to obtain a comprehensive understanding of how the signal transduction pathways that control the SPI1 T3SS are integrated during the infection process. Extensive genetic analysis has allowed us to formulate a new model for the SPI1 regulatory circuit in which the three AraC-like regulators HilD, HilC, and RtsA act in a complex feed-forward regulatory loop to control expression of hilA, encoding the direct regulator of the SPI1 structural genes. We hypothesize that regulatory signals feed into the system primarily via post-translational control of HilD, which in turn activates hilC, rtsA, and hilA. But how these regulatory systems control HilD is unknown. The flagellar protein FliZ and the protein HilE independently control HilD at the protein level, most likely via protein-protein interaction with the N-terminal domain of HilD. As these represent proximal regulatory inputs, we focus on understanding how FliZ and HilE control HilD function. The specific aims of this proposal are to 1. Determine how FliZ and HilE act to control HilD activity. Biochemical and genetic experiments will dissect each step in HilD activation of hilA to determine how FliZ and HilE act to control HilD function or stability. 2. Characterize the nature of the interaction between HilD and HilE or FliZ. Co-immunoprecipation and two-hybrid analysis will be used to characterize HilE-HilD and FliZ-HilD interactions. HilD point mutations that negate regulation will be used to identify regions of HilD that are specifically required for HilE- or FliZ-dependent regulation. These mutants will also allow us to test the role of FliZ- and HilE-dependent regulation of SPI1 during intestinal invasion. 3. Determine the signal transduction pathways that feed into HilD to control the SPI1 T3SS. Known regulatory systems that control SPI1 T3SS expression will be screened for those that function through HilD. Mass spectrometry and two-hybrid analysis will be used to identify additional factors that interact with HilD protein. Characterization of these factors will lead to our overall understanding of global signal transduction. The regulation of the SP1 T3SS serves as a paradigm for the integration of host environmental signals to control virulence gene expression and analysis of this system is critical to our understanding of this Class B priority pathogen. Salmonella are major food-borne pathogens in the US. The bacteria invade the human intestinal cells to cause disease. Our goal is to understand the regulation of the bacterial invasion system to improve prevention and/or treatment

Keywords: Affect; Amplifiers; Bacteria; base; Biochemical Genetics; Biosensor; Cause of Death; Cells; Co-Immunoprecipitations; Complex; Diarrhea; Disease; Epithelial Cells; feeding; foodborne; foodborne pathogen; Gastroenteritis; Gene Expression; Genetic; genetic analysis; Genetic Transcription; Goals; Half-Life; Human; improved; Individual; Infection; Inflammatory; Injection of therapeutic agent; Intestinal Diseases; Intestines; Invaded; Knowledge; Lead; Mass Spectrum Analysis; Measures; Mediating; Modeling; mutant; Mutation; N-terminal; Nature; novel; pathogen; Pathogenesis; Pathogenicity Island; Point Mutation; Prevention; Process; protein protein interaction; Proteins; public health relevance; Regulation; Research; research study; response; Role; Salmonella; Salmonella enterica; Signal Transduction; Signal Transduction Pathway; Site; SP1 gene; Structural Genes; System; Systemic disease; Systemic infection; Systems Analysis; Testing; Type III Secretion System Pathway; Typhoid Fever; Virulence; Work; yeast two hybrid system

Relevance: Salmonella are major food-borne pathogens in the US. The bacteria invade the human intestinal cells to cause disease. Our goal is to understand the regulation of the bacterial invasion system to improve prevention and/or treatment

Project start date: 2010-07-15

Project end date: 2014-06-30

Budget start date: 1-JUL-2011

Budget end date: 30-JUN-2012

PFA/PA: PA-07-070

5R01AI080705-02 (2011): $362891

PRECLINICAL STUDIES OF AAV GENE THERAPY IN MOUSE MODELS OF UREA CYCLE DISORDERS

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Abstract: PROJECT II - PRECLINICAL STUDIES OF AAV GENE THERAPY IN MOUSE MODELS OF UREA CYCLE DISORDERS AND IN NONHUMAN PRIMATES The goal of this project is to evaluate the potential of an optimized clinical candidate AAV vector (AAVcc) developed in Project I, for efficacy, duration and safety as a potential therapeutic vector in murine models of urea cycle disorders and in neonatal nonhuman primates (NHP). This project builds upon our recent success with the use of a novel AAV serotype, AAV8, in protecting ornithine transcarbamylase (OTC) deficient adult spf and spfash mice from hyperammonemia. Specific Aim 1 will evaluate the AAVcc in the treatment of adult spfash mice. We will determine how rapidly protection from an ammonia challenge is conferred after gene transfer of OTC and the duration of metabolic stability. Specific Aim 2 will evaluate the potential of gene therapy in younger recipients. Initial studies will be performed with AAVcc expressing the reporter gene GFP, administered to wild-type mice at various stages following birth, from 1 day to 4 weeks of age. Animals will be harvested at various times to measure the rate of onset of transgene expression. Additional cohorts will be followed over time to assess stability of AAV-mediated gene transfer when administered into young animals. These experiments will define important parameters to further explore the potential of the clinical candidate in treating young spfsh animals. The most stringent test for the clinical candidate will be performed in animals completely deficient in OTC through targeted gene disruption (OtcKO). This OtcKO line will be generated during the early phase of the project. If there is a delay in generating this KO model, we will use as a surrogate, the existing argininosuccinate synthase (AS) KO. All studies performed in the spfsh and OTC/AS KO animals will also involve measures of safety, including a series of clinical chemistry and hematology measurements as well as histopathology of tissues harvested and necropsied. The parameters of safety and efficacy defined in Specific Aim 2 will be further evaluated in neonatal NHP studies in Specific Aim 3. Newborn cynomolgus macaques will be injected with AAVcc expressing cynomolgus-derived OTC cDNA. Animals will be necropsied subsequent to gene transfer and evaluated for 1) gene transfer by fluorescent in situ hybridization; 2) safety; 3) histopathology and clinical chemistry; 4) and T cells directed against the vector capsid. The final specific aim will evaluate the potential role of specific human OTC mutations that could interfere with the success of gene therapy. Existing mutations may interfere with the activity of the product of the normal transgene through a dominant negative mechanism. We will coexpress mutant and wild-type OTC in CHO cells to evaluate mutations that have these effects. Mutants that appear to show dominant negative effects in vitro will be selected for further examination in vivo using AAV gene delivery of the mutant OTC into wild-type mice. Together, these studies will allow us to determine the effectiveness of the clinical candidate vector for use in gene therapy. Lay description. In Project II, the clinical candidate vector, developed in project I, will be assessed for efficacy and safety in animal models

Keywords: Acute; adeno-associated viral vector; Adult; Age; Ammonia; Animal Model; Animals; Arginine; argininosuccinate synthase; Autopsy; Biochemical; Biology; Birth; Blood specimen; Capsid; Cell membrane; Cells; Chinese Hamster Ovary Cell; Clinical; Clinical Chemistry; Clinical effectiveness; Clinical Immunology; Clinical Pathology; clinically significant; cohort; Complementary DNA; Disease; Dominant-Negative Mutation; Engineering; enzyme activity; Enzymes; Fluorescent in Situ Hybridization; Follow-Up Studies; Gene Delivery; Gene Expression; Gene Targeting; gene therapy; Gene Transfer; Genome; Glutamine; Goals; Growth; Harvest; Hematology; Histocytochemistry; Histopathology; Human; Hyperammonemia; In Situ; In Vitro; in vivo; Infusion procedures; juvenile animal; Life; Liver; Macaca; Measurement; Measures; Mediating; Metabolic; Modeling; Molecular; mouse model; Mus; mutant; Mutation; Neonatal; Newborn Infant; nonhuman primate; novel; Ornithine Carbamoyltransferase; Ornithine carbamoyltransferase deficiency; orotate; Patients; peripheral blood; Phase; Phenotype; Plasma; preclinical study; pregnant; prevent; Principal Investigator; programs; reconstitution; Reporter Genes; research study; response; Role; Safety; Sampling; Series; Serotyping; Speed (motion); Staging; success; T-Lymphocyte; Testing; Therapeutic; Time; Tissue Harvesting; transgene expression; Transgenes; urea cycle; urinary; vector; Viral Genome; Wild Type Mouse

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5P01HD057247-04_0002 (2011): $146894

IMMUNE BARRIERS TO AAV GENE THERAPY

M James, Professor A
University Of Pennsylvaniacity: Philadelphia    country: United States (us)

Abstract: PROJECT I - IMMUNE BARRIERS TO AAV GENE THERAPY The performance of novel AAV serotypes, with the use of the self complementing genome, has vastly improved the prospects of successful liver-directed in vivo gene therapy. Despite these encouraging data, a number of potential barriers remain, primarily focused on the immunologic biology of in vivo gene therapy. This project will systemically address the immunologic response to in vivo gene therapy of novel AAVs as a prerequisite to their considerations in clinical applications. The first specific aim will focus on the identification of a clinical candidate which is defined as the actual vector to be considered in the Phase 1 clinical trial. The two components of the vector that will be extensively studied and optimized are 1) the capsid, evaluated for efficiency and stability of gene transfer, toxicity, transgene and capsid T cells, pre-existing immunity and biodistribution; and 2) the genome, evaluated for peak and onset of expression. The second specific aim will analyze the role of pre-existing T cells to AAV capsids in terms of the safety and efficacy of liver-directed gene transfer. The third specific aim will evaluate the role of the target organ in eliciting problematic immunologic responses, specifically focusing on activation of innate immunity or inflammation. These studies will focus on murine systems in establishing basic principles which are followed up selectively in nonhuman primates. The project will extensively use the Vector and Morphology Cores and will collaborate directly with Project II on the evaluation of vector efficacy in the OTC-deficient mouse model and with Project III by providing NHP tissue for molecular characterization. Lay description. A vector of use for the treatment of OTC deficiency called the clinical candidate will be created. Potential immunologic responses of the recipient to the vector will be studied

Keywords: Address; adeno-associated viral vector; Adenoviruses; Animals; Antigen Targeting; base; Biodistribution; Biology; Capsid; Cells; Clinical; clinical application; Clinical Research; Clinical Trials; Complement; Complementary DNA; Data; Dependovirus; Disease; Enrollment; Evaluation; Family; follow-up; gene therapy; Gene Transfer; Generations; Genome; Hepatocyte; Human; Immune; immune activation; Immunity; Immunization; Immunologics; improved; in vivo; Infection; Inflammation; Interferons; Liver; Macaca; mature animal; Memory; Methods; Molecular; Morphology; mouse model; Mus; Mutation; Natural Immunity; nonhuman primate; novel; Open Reading Frames; Organ; Ornithine Carbamoyltransferase; Ornithine carbamoyltransferase deficiency; Pathway interactions; Performance; Phase I Clinical Trials; Population; pre-clinical; preclinical study; Predisposition; Primates; Process; Promotor (Genetics); Research; research study; response; Risk; Role; Safety; Series; Serotyping; System; T memory cell; T-Cell Activation; T-Lymphocyte; Tissues; TLR3 gene; Toxic effect; transgene expression; Transgenes; urea cycle; vector; vector genome

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5P01HD057247-04_0001 (2011): $146894

OBESITY: FACTORS INVOLVED IN GENESIS OF PREECLAMPSIA

M James, Professor
Magee-women´s Res Inst And Foundationcity: Pittsburgh    country: United States (us)

Abstract: Preeclampsia is more common in obese women. In our population the risk is 3 times greater than that of lean women. The frequency of obesity (25-30%) results in an attributable risk for preeclampsia of 35 to 50%. This increased risk is not only for mild but also for severe and early onset preeclampsia. This frequency of obesity is similar in developed countries and obesity is becoming more common in developing counties. It is important to understand the mechanism(s) by which obesity increases the risk of preeclampsia. A key question what factor(s) contribute or cause a particular obese woman to develop preeclampsia? We will ask whether percent body fat, or distribution or accrual of fat, are related to preeclampsia. We will also study metabolic and inflammatory changes, known to be present in both preeclampsia and obesity, testing the hypothesis that obese women who develop preeclampsia will manifest more abnormalities in these factors in early pregnancy. We will also test whether periconceptional diet, sleep disorders or smoking are different in obese women who develop preeclampsia. These questions will be tested primarily in a longitudinal study of 600 obese nulliparous women sampled at 8-10, 18-20, 34-36 weeks gestation and at delivery. Because severe preeclampsia is rare, we will examine the metabolic, inflammatory and angiogenic changes in a cross sectional study of 25 obese and 25 lean women, with or without severe preeclampsia. Our preliminary data suggest that asymmetric dimethylarginine (ADMA), the endogenous inhibitor of the interaction of L-arginine with nitric oxide synthase (NOS), is increased in early pregnancy in obese women who subsequently develop preeclampsia. All of the factors we are studying are known to increase ADMA and we will test whether these factors converge to increase ADMA with subsequent vascular dysfunction. We will test this by measuring and correlating ADMA with the other factors but also by administering L-arginine to 30 women between 16 and 19 weeks gestation, testing vascular dysfunction, metabolic and inflammatory markers before and after treatment in a doubly masked randomized controlled trial. The goal of Project II is to identify modifiable factors in obese women to prevent preeclampsia and to determine if there is a common target, overcoming the effect of ADMA to inhibit NOS that would be useful to prevent preeclampsia

Keywords: Address; Aftercare; Arginine; Biological; Blood Vessels; Body fat; Cardiovascular Diseases; cohort; Country; County; Cross-Sectional Studies; Data; Diabetes Mellitus; Diet; Disease; Dyslipidemias; early onset; Fatty acid glycerol esters; Frequencies (time pattern); Functional disorder; Goals; Growth Factor; Hypertension; improved; in vivo; Individual; Inflammation; Inflammatory; inflammatory marker; inhibitor/antagonist; Insulin Resistance; Lead; lifestyle factors; Lipids; Longitudinal Studies; Masks; Measures; Metabolic; metabolic abnormality assessment; Metabolic Marker; Metabolic syndrome; N, N-dimethylarginine; Nitric Oxide Synthase; Obesity; obesity risk; Oxidative Stress; Population; Pre-Eclampsia; Pregnancy; Pregnant Women; prevent; Randomized Controlled Trials; receptor; Recruitment Activity; Risk; Sampling; Sleep Disorders; Smoking; Testing; Time; Woman

Budget start date: 1-APR-2011

Budget end date: 31-MAR-2012

5P01HD030367-17_0015 (2011): $222415

SIDEPORT NEEDLE ARRAY TECHNOLOGIES FOR PRIORITIZING DRUGS FOR CANCER PATIENTS

M James, Associate Member
Presage Biosciences, Inc.city: Seattle    country: United States (us)

Grant 4R42CA144104-02 from National Cancer Institute

Abstract: Over 90% of cancer patients that enroll in Phase I or II clinical trials experience no benefit from the experimental therapies, yet are exposed to drug toxicity and other challenges related to treatment. For over 50 years, physicians have used patient-specific information about drug resistance and sensitivity to select antibiotics for patients with infections, but this personalized approach has evaded the oncology community because cancer cell behavior in vitro drug sensitivity assays does not correlate with in vivo response to therapy in most cases. We have developed technologies that enable oncology drug sensitivity/resistance testing of multiple drugs or drug combinations in vivo during the days prior to surgical resection of a tumor. This approach allows drugs to interact with cancer cells while the latter are in their native tumor microenvironment. Our broad long-term goal is to develop reliable in vivo-based oncology drug sensitivity/resistance assays for patients with many types of solid tumors. Our overall goal of the STTR Phase I and II projects are to develop and test devices that are suitable for human lymphoma patients and to initiate human clinical trials. Our Specific Aim for the Phase I portion is to develop a single use (disposable) porous needle array and demonstrate that it meets drug delivery precision specifications. Provided that quantitative milestones are met in Phase I, Phase II will proceed with the following Aims Aim 1) To develop a prototype suitable for use in human lymphoma patients; and Aim 2) to conduct a pilot "first in humans" clinical trial. The significance of the proposed work is that it will reduce the frequency of cancer patients being exposed to drugs that cause toxicity but offer no clinical benefit. The commercialization potential is described in a comprehensive business plan. We provide letters from highly respected individuals in the biotechnology, life sciences and personalized medicine fields to attest to the commercial potential of this technology. It is estimated that approximately 1.4 million new cases of cancer will be diagnosed in the United States in 2008. Improved methods for prioritizing cancer therapeutics based on patient-based indicators of efficacy are needed. We are proposing to develop a device which enables comparison of multiple drugs or combinations in vivo, with the tumor micro-environment intact. The long-term goal of this research is to develop reliable in vivo- based oncology drug sensitivity/resistance assays for patients with many types of solid tumors. This technology will reduce the frequency of cancer patients being exposed to drugs that cause toxicity but offer no clinical benefit. This personalized treatment approach will improve patient outcome and enhance the quality of care for millions of individuals that suffer from cancer

Keywords: Address; Antibiotics; base; Biological Assay; Biological Sciences; Biotechnology; Businesses; cancer cell; Cancer Model; Cancer Patient; Caring; cell behavior; chemotherapy; Clinical; Clinical Data; clinical practice; Clinical Trials; cohort; commercialization; Community Clinical Oncology Program; Coupled; Data; Devices; Diagnosis; drug candidate; Drug Combinations; Drug Delivery Systems; Drug resistance; drug sensitivity; Drug toxicity; Engineering; Enrollment; Environment; Excision; experience; Foundations; Frequencies (time pattern); Funding; Goals; Grant; Heterogeneity; Hour; Human; human data; improved; In Vitro; in vivo; Individual; Infection; Investigational Therapies; Journals; Letters; Lymphoma; Malignant Neoplasms; Manuscripts; Marketing; Medical Device; Medical Oncologist; Medicine; meetings; Methods; Mus; Nature; Necrosis; Needles; oncology; Operative Surgical Procedures; Outcome; Pathology; Patients; Pattern; Peer Review; Pharmaceutical Preparations; Phase; Phase I/II Trial; Physicians; Play; pre-clinical; prototype; public health relevance; Publishing; Quality of Care; Research; Research Personnel; Resistance; response; Role; Running; Science; Scientist; Small Business Technology Transfer Research; Solid Neoplasm; Technology; Testing; Therapeutic; Toxic effect; tumor; United States; Work

Relevance: It is estimated that approximately 1.4 million new cases of cancer will be diagnosed in the United States in 2008. Improved methods for prioritizing cancer therapeutics based on patient-based indicators of efficacy are needed. We are proposing to develop a device which enables comparison of multiple drugs or combinations in vivo, with the tumor micro-environment intact. The long-term goal of this research is to develop reliable in vivo- based oncology drug sensitivity/resistance assays for patients with many types of solid tumors. This technology will reduce the frequency of cancer patients being exposed to drugs that cause toxicity but offer no clinical benefit. This personalized treatment approach will improve patient outcome and enhance the quality of care for millions of individuals that suffer from cancer

Project start date: 2011-05-01

Project end date: 2013-04-30

Budget start date: 1-MAY-2011

Budget end date: 30-APR-2012

PFA/PA: PA-09-081

4R42CA144104-02 (2011): $544375

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