THE WOMEN AND [THEIR] CHILDREN´S GULF HEALTH CONSORTIUM
J Edward, Associate Dean For Research
Louisiana State Univ Hsc New Orleanscity: New Orleans country: United States (us)
Grant 1U01ES021497-01 from Office Of The Director, National Institutes Of Health
Keywords: Budgets; Child; cohort; Funding; Grant; Health; Interview; outreach; Sample Size; Woman; Work
Project start date: 2011-06-27
Project end date: 2016-04-30
Budget start date: 27-JUN-2011
Budget end date: 30-APR-2012
PFA/PA: RFA-ES--1-006
1U01ES021497-01 (2011): $340591
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to J Edward
PROTEIN 4.1 GENE EXPRESSION IN DEVELOPING RED CELLS
J Edward, Professor & President
Dana-farber Cancer Institutecity: Boston country: United States (us)
Grant 5R01HL024385-33 from National Heart, Lung, And Blood Institute
Abstract: We have employed the protein 4.1R gene as a model to study the molecular regulation of splicing during erythropoiesis. Protein 4.1R is a key element of the erythrocyte cytoskeleton. Two tightly regulated splicing events alter its expression and function during erythropoiesis 1) An early event dictates exon 2´ exclusion, which omits an upstream translation initiation site and produces only the "small" (80 kD) isoform. 2) A late event induces the inclusion of exon 16, which encodes a peptide critical for spectrin-actin binding. We have made progress toward understanding both events, including 1) Identification of a stage-specific mFox-2A isoform that is up-regulated in late erythroid cells and critical for the exon 16 splicing switch; 2) Demonstration that increased SF2/ASF expression in late erythroid cells stimulates exon 16 splicing by binding an exonic splicing enhancer; 3) Quantification of a panel of splicing factors during erythroid differentiation, showing 4.1R splicing is modulated by varying the amount of selected ubiquitous and cell-type specific activators and inhibitors; two of these, PTB and a 40 kD novel protein are objects of the proposed studies; 4) Identification of a coupled transcription and splicing pathway that regulates exon 2´ splicing; and 5) Detection of a switch in promoter usage that results in the production of the 80 kD isoform in mature red cells. We now propose to define more precisely the key elements and mechanisms governing these two splicing events, and to establish the physiological role of the mFox-2A isoform that we discovered, employing the following specific aims 1) To complete the characterization of the functional roles of PTB and a 40 kD protein in exon 16 splicing regulation and to characterize the mechanisms by which they promote exon 16 splicing activation. This will employ combined genetic and biochemical approaches in both in vitro and in vivo systems. 2) To determine the physiological impact of Fox-2A in directing erythroid specific alternative exon selection. The impact of Fox-2A will be analyzed by comprehensive analysis of erythroid specific splicing changes that occur in late cells, using SpliceArray technologies. 3) To investigate how transcription from the alternative exon 1A and 1C promoters governs exon 2´/2 splice site selection. These studies will provide insights into how particular factors function to program the differentiation of erythroid cells. Many different forms of a protein can be generated from a gene by alternative mRNA splicing; improper production of these protein forms could have a profound impact on human diseases. We study a major example (Protein 4.1R) of how this splicing is regulated
Keywords: 3` Splice Site; acrosome stabilizing factor; Actins; Affect; Alternative Splicing; Amanitins; base; Behavior; Binding (Molecular Function); Binding Sites; Biochemical; Biochemical Genetics; Biogenesis; C-terminal; Cell membrane; cell type; Cells; CFU-E; Complex; Coupled; crosslink; Cytoskeleton; design; Detection; Development; Distal; DNA Polymerase II; Elements; Enhancers; Erythroblasts; Erythrocyte Membrane; Erythrocytes; Erythroid; Erythroid Cells; erythroid differentiation; Erythropoiesis; Event; Exclusion; Exons; Foxes; gain of function; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Glycophorin; Goals; Health; human disease; In Vitro; in vivo; inhibitor/antagonist; insight; Introns; Lead; Limb structure; loss of function mutation; Mediating; membrane biogenesis; Membrane Proteins; Messenger RNA; Methods; Modeling; Molecular; mRNA Precursor; mutant; novel; Pathway interactions; Pattern; Peptides; Physiological; Play; Production; programs; Promotor (Genetics); protein 4.1; Protein Isoforms; Proteins; prototype; Recruitment Activity; Regulation; Resistance; RNA Interference; RNA Splicing; Role; Site; Spectrin; Staging; System; Technology; Testing; Tetanus Helper Peptide; Trans-Activators; Translation Initiation; Translations; vector
Project start date: 1995-01-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-07-070
5R01HL024385-33 (2011): $427500
VIRGINIA TECH POST-BACCALUAREATE RESEARCH AND EDUCATION PROGRAM
J Edward, Dr
Virginia Polytechnic Inst And St Univcity: Blacksburg country: United States (us)
Grant 5R25GM066534-08 from National Institute Of General Medical Sciences
Abstract: Virginia Tech, the largest research university in the Commonwealth of Virginia, proposes to continue its successful Virginia Tech Post-baccalaureate Research and Education program (VT-PREP) for another four years by recruiting and preparing a total of 29-32 scholars for admissions into highly reputable and productive biomedical and behavioral sciences doctoral programs at Research I universities. In the last four years, 75% of the 28 scholars who participated in the VT-PREP were accepted into highly selective doctoral programs including University of Rochester, Virginia Tech, University of Texas, and Purdue University. The current training plan will include a combination of educational, experiential, and social activities in a supportive environment that ensures development of each scholar both academically and socially. Specific objectives for this renewal include 1. Recruit and enroll 29-32 minority students underrepresented in the biomedical and behavioral sciences to participate in a 12-month transition program as research scholars 2. Develop and implement activities and programs that will enrich and motivate scholars as well as facilitate the pursuance of a research career in the biomedical sciences. 3. Increase the number of minority applicants applying for and being accepted into graduate studies in biomedical, agricultural, and life sciences and related disciplines at Virginia Tech and improving the campus climate for diversity. Each VT-PREP scholar will participate in enrichment activities including seminars, gateway courses, and workshops. Additionally, 75 to 80% of each scholar´s time will be devoted to a mentored research activity under the guidance of VT faculty with strong research, teaching, and mentoring accomplishments in the biomedical and behavioral sciences. We anticipate the following quantifiable and qualitative outcomes from implementation of the VT-PREP objectives 1) An increased proportion of applications from minority applicants to pursue graduate education in the biomedical and behavioral sciences at VT 2) An increased awareness of Virginia Tech graduate programs and the positive environment the university has developed for minorities; 3) 75 to 100% of the VT-PREP scholars will pursue graduate studies in biomedical and behavioral sciences at VT and other major research institutions. The impact of PREP on VT has included helping research-intensive labs recruit and train minorities, developing a seminar series that involves reflections by scientists on their journey in science, and exposing students and faculty at HBCUs to research opportunities at VT. VT-PREP will continue to be a valued institutional partnership between the central administration, the Graduate School and several colleges. It will be led by a three-member team of accomplished scientists with experience in training students including under represented minorities at all levels. The PREP will be used to train underrepresented minorities in the biomedical and behavioral sciences to become competitive for admission into highly competitive doctoral research programs
Keywords: Admission; Admission activity; agricultural; Agriculture; Authorship; Awareness; Awarenesses; Behavioral Sciences; Biologic Sciences; Biological Sciences; Biomedical Research; career; Climate; climatic; cohort; college; Commit; conference; Development; Discipline; Doctor of Philosophy; Education; Educational aspects; Educational process of instructing; Educational workshop; enroll; Enrollment; Ensure; Environment; experience; Faculty; Goals; Graduate Education; graduate student; HBCUs; Historically Black Colleges; Historically Black Colleges and Universities; Historically Black Institution; Hour; improved; Institution; Intervention; Intervention Strategies; interventional strategy; Interview; Life Sciences; meetings; member; Mentors; Meteorological Climate; Minority; National Human Genome Research Institute; National Institute of General Medical Sciences; NCI Scholars Program; NIGMS; Outcome; outreach; pathway; Pathway interactions; Peer Review; Ph.D.; PhD; Principal Investigator; programs; Programs (PT); Programs [Publication Type]; R01 Mechanism; R01 Program; recruit; Recruitment Activity; Research; Research Activity; Research Grants; Research Project Grants; Research Projects; Research Projects, R-Series; RPG; Scholars Program; Schools; Science; Scientist; Series; social; Social Network; Students; success; symposium; Teaching; Texas; Time; Training; Training Programs; under-represented minority; Underrepresented Minority; underserved minority; Universities; Virginia; Woman; Workshop
Project start date: 2002-09-01
Project end date: 2013-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PAR-07-432
5R25GM066534-08 (2011): $249928
J Edward
University Of North Carolina Chapel Hillcity: Chapel Hill country: United States (us)
Abstract: The Vector Labs are structured and organized under a new paradigm for Vector Core operations that is designed to provide investigators with a single source that can supply their vector needs throughout the development of the research programs. The labs utilize an integrated systems approach to speed the flow of vector technologies and information. These systems are designed to provide comprehensive resources to investigators so they can avoid many of the common pitfalls and delays that occur during the translational research period. These pitfalls and delays are caused when a specific vector-disease approach must transition from a basic research environment into the highly regulated and controlled processes of final preclinical safety testing and human clinical trials. The UNC Vector Labs will support MTCC investigators by providing expertise with vector design, construction, and development as well as by providing investigators with the viral vector reagents themselves. Investigators will be able to select the optimal type of viral platform (adenoviral, AAV, lentiviral, retroviral, PIV, cell-based, or plasmid) for the specific research in which they are involved. Investigators can also select from a range of standard quality grades (research, preclinical, or clinical), or can custom design intermediate grades as necessary. The combined staff at the Vector Labs and the Gene Therapy Center have prior experience in bringing to clinical trials virtually all the major viral platforms. Investigators will be able to utilize this experience to aid in creating drug development templates as their research progresses into the clinical arena
Keywords: adeno-associated viral vector; Adenoviruses; base; Basic Science; Cells; Clinical; Clinical Trials; Communicable Diseases; Custom; cystic fibrosis mouse; design; design and construction; Development; Disease Vectors; drug development; Environment; experience; gene therapy; Gene Transfer; gene transfer vector; General Population; Hereditary Disease; Human; Immunization; Information Technology; intestinal epithelium; Lentivirus Vector; Mission; Molecular; Mutation; Neonatal; operation; Plasmids; pre-clinical research; preclinical safety; Primary Ciliary Dyskinesias; Process; Production; programs; public health medicine (field); Reagent; Research; research and development; Research Personnel; Resources; Retroviridae; safety testing; Services; small hairpin RNA; Small Interfering RNA; Source; Speed (motion); Structure; Subfamily lentivirinae; System; Training; Training Programs; Transgenes; Transgenic Mice; Translational Research; vector; Viral; Viral Vector
Relevance: The Vector Core will provide a broad range of services that will benefit public health. Its foremost mission is to provide gene transfer vectors that will treat people with genetic diseases. An important parallel mission is to develop gene transfer vectors that may aid in immunization of the general population to a variety of communicable diseases
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5P30DK065988-08_7435 (2011): $143418
NHERF-1 AND PTH REGULATION OF THE RENAL TRANSPORT OF PHOSPHATE.
J Edward, Professor
University Of Maryland Baltimorecity: Baltimore country: United States (us)
Grant 5R01DK055881-13 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: The Sodium-Hydrogen Exchanger Regulatory Factor-1 (NHERF-1) is an adaptor protein containing two protein-interactive PDZ domains and a C-terminal ERM binding domain that localizes to the brush border membrane of renal proximal convoluted tubule cells and binds to Npt2a, the major sodium-dependent phosphate transporter. Our recent studies have indicated that sodium-dependent phosphate transport in proximal tubule cells from NHERF-1-/- kidneys are resistant to the inhibitory effect of Parathyroid Hormone (PTH). In this current application, we explore the hypothesis that PTH mediates the phosphorylation of specific residues in PDZ I of NHERF-1 thereby regulating Npt2a/NHERF-1 complexes, the abundance of Npt2a in the apical membrane of renal proximal tubule cells, and as a consequence, the tubular reabsorption of phosphate. Elucidation of the factors that regulate the binding of target proteins to PDZ I of NHERF-1 may also provide broader insights into how regulation of PDZ domains of adaptor proteins impact on biologic responses to hormones and on the pathophysiology of NHERF-1 related diseases. In intact animals and cultured proximal tubule cells, we will use physiologic, biochemical, and cell biologic assays to determine how PTH-mediated phosphorylation of PDZ I of NHERF-1 regulateS the binding affinity of target proteins such as Npt2a and the proximal tubule reabsorption of phosphate. We propose three specific aims. First, we will map the serine and/or threonine residues in PDZ I of NHERF-1 that are phosphorylated in response to PTH and downstream protein kinases. Second, we propose to study the association and dissociation of Npt2a/NHERF-1 complexes in response to PTH-mediated phosphorylation of NHERF-1 using in-vitro and in-vivo assays. Third, we will determine the physiologic role of PTH-mediated NHERF-1 phosphorylation on the regulation of phosphate transport in the proximal tubule of the kidney. PUBLIC HEALTH RELEVANCE NHERF-1 is an adaptor protein that binds multiple transporters in the kidney including Npt2a, the major renal proximal tubule phosphate transporter. We will study the hypothesis that Parathyroid Hormone-mediated regulation of renal phosphate transport involves regulation of the binding of Npt2a to NHERF-1 by site-specific phosphorylation of the PDZ I domain of NHERF-1. These observations may provide mechanistic insights into the processes that regulate the binding of target proteins to adaptors such as NHERF-1 and provide broader insights into the pathophysiology of NHERF-1 related diseases
Keywords: 8-Bromo Cyclic Adenosine Monophosphate; Accounting; Adaptor Signaling Protein; Adenoviruses; Affinity; Animals; apical membrane; Binding (Molecular Function); Binding Proteins; Biochemical; Biological Assay; brush border membrane; C-terminal; Cell Line; Cell membrane; Cells; Co-Immunoprecipitations; Complex; Confocal Microscopy; Cyclic AMP-Dependent Protein Kinases; Cytoskeleton; Density Gradient Centrifugation; Diet; Disease; Dissociation; Elements; Estrogen receptor positive; Excretory function; ezrin; Family; Functional disorder; Gel; Genes; Goals; hormone regulation; Hormones; human PTH protein; In Vitro; in vivo; Ingestion; inorganic phosphate; Inorganic Phosphate Transporter; insight; interest; Ion Channel; Kidney; malignant breast neoplasm; Maps; Mediating; member; Membrane; Mitogen-Activated Protein Kinases; Modeling; moesin; Mus; Mutation; Nephrolithiasis; Neurofibromin 2; NHERF-1 protein; Parathyroid Hormone Receptor; Pathway interactions; Phosphoproteins; Phosphorylation; Phosphorylation Site; Physiological; Process; Protein Binding; Protein Dephosphorylation; Protein Family; Protein Hormone Receptor; Protein Kinase; Protein Kinase C; Proteins; Proximal Kidney Tubules; Psoriasis; public health relevance; radixin protein; Regulation; Reporting; Resistance; response; Role; Scaffolding Protein; Schizophrenia; second messenger; Second Messenger Systems; Serine; Signal Transduction; Signaling Protein; Site; Site-Directed Mutagenesis; Slice; Sodium; sodium-hydrogen exchanger regulatory factor; Stimulus; Sucrose; Surface Plasmon Resonance; Threonine; trafficking; Tubular formation; urinary
Project start date: 1999-05-01
Project end date: 2013-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01DK055881-13 (2011): $312407
J Edward
University Of Miami School Of Medicinecity: Miami country: United States (us)
Abstract: The purpose of the behavioral Core is to evaluate the functional status of neural circuitry following traumatic brain injury in rats and mice, by quantitatively assessing sensorimotor and cognitive behavioral endpoints including seizures. Paradigms are designed to assess neurobehavioral deficits associated with fluid percussive TBI in rats, and controlled cortical impact in mice, and to evaluate, when possible, the relation between neurobehavioral endpoints and neuropathological, and neurophysiological markers. The tests of sensorimotor integration and cognition used in the core are have been demonstrated to be sensitive to insults produced by these TBI models, which, at moderate to severe levels of injury severity, produce overt pathology in the parietotemporal cortex, hippocampus, thalamus, and white matter
Keywords: Attention; Attenuated; base; Behavior; Behavior assessment; behavior measurement; behavior test; Behavioral; Cognition; Cognitive; controlled cortical impact; Data; design; Development; environmental enrichment for laboratory animals; Ephrins; Financial compensation; Forelimb; functional outcomes; functional status; Gait; Hippocampus (Brain); improved; injured; Injury; kinematics; Learning; Liquid substance; Maintenance; Modeling; Mus; nerve stem cell; neural circuit; neurobehavioral; neurophysiology; neurotrophic factor; Pathology; Play; Posture; programs; Rattus; receptor; Recovery; Rehabilitation therapy; repaired; research study; Role; Seizures; Severities; Synapses; synaptogenesis; Testing; Thalamic structure; Therapeutic; Therapeutic Intervention; therapy design; Training; Transgenic Mice; Transplantation; Traumatic Brain Injury; treatment strategy; white matter
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
5P50NS030291-18_5440 (2011): $155300
NMR AND NIR DETECTION OF PHOSPHOLIPASE ACTIVITY IN VIVO
J Edward, Research Associate Professor
University Of Pennsylvaniacity: Philadelphia country: United States (us)
Grant 5R01CA129176-03 from National Cancer Institute
Abstract: The overall aim of this application is to develop methods to image the expression and activity of phospholipases in vivo. A multi-modality approach with sequential magnetic resonance imaging/spectroscopy (MRI/MRS) and optical imaging using near infrared (NIR) fluorophores will be employed to obtain information that is not available from either technique separately. The key hypothesis to be addressed is that in vivo activation of phospholipases can be measured by determining the changes in MR-visible lipid metabolite levels together with measuring fluorescence released by enzymatic hydrolysis of a self- quenching NIR phospholipid fluorophore. We will compare and validate the MR spectroscopy with results obtained using NIR optical imaging in tumor models in which changes in choline metabolite levels associated with tumor progression and response to therapy are well known. Since phospholipases have been implicated in a wide range of pathologies including cancer, arthritis and Alzheimer´s disease, this study will supply critical in vivo data on the specific biochemical pathways involved in these diseases. The hypothesis will be tested by the achievement of the following specific aims Aim 1) Design and synthesis of chiral lipid-based self-quenching fluorophores containing a stable pyropheophorbide NIR fluorophore, coupled to a dark quencher. The fluorophore will be attached to different positions of the glycerol backbone to interrogate the activity of different phospholipase isoforms. The quencher linkage will be attached at different points on the glycerol backbone to distinguish between phospholipase types. Structure-activity studies will be performed by synthetically inserting linkers of various length to determine the effects of steric hindrance on probe activity. Aim 2) In vitro characterization of the synthesized probes. This will include test-tube demonstration of the sensitivity and specificity of the probes to various phospholipase types and isoform and detailed measurement of kinetics of catalysis and inhibition. We will further characterize the delivery and activation of these self- quenching fluorophores in cultured cell lines. Aim 3) Demonstration of in vivo delivery, bio-distribution and subsequent activation of phospholipases in tumor xenograft models of prostate cancer and non-Hodgkin´s lymphoma, followed by dual modality molecular imaging using 1H MR spectroscopy and optical imaging. These data will supply critical information on the specific lipid catabolic pathways involved in the onset and subsequent treatment of disease. To develop near-infrared fluorescent beacons for the in vivo detection of phospholipase activity via dual optical imaging and MR spectroscopy
Keywords: Achievement; Address; Alzheimer`s Disease; Animals; Antineoplastic Agents; Arthritis; Autoimmune Process; bacteriochlorin; base; Biochemical; Biochemical Pathway; biological systems; Biomedical Engineering; Cancer cell line; Catalysis; Cell Line; Cells; Characteristics; Choline; clinical Diagnosis; Coupled; Data; design; Detection; Development; Disease; Early Diagnosis; Enzymes; Equilibrium; Event; Fluorescence; fluorophore; Glycerol; Goals; Health; Human; Hydrolysis; Image; imaging modality; imaging probe; Imaging Techniques; improved; In Situ; In Vitro; in vivo; Inflammatory; Kinetics; Length; lipid metabolism; Lipids; Magnetic Resonance; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Malignant neoplasm of prostate; Malignant Neoplasms; Measurement; Measures; Metabolic; Metabolism; Methods; Modality; Modeling; Molecular; molecular imaging; Monitor; Non-Hodgkin`s Lymphoma; optic imaging; Organ; Pathology; Pathway interactions; Pharmacotherapy; phosphodiester; phosphoethanolamine; Phospholipase; Phospholipase A2; Phospholipase C; Phospholipids; phosphomonoester; Phosphorylcholine; Physiological; Positioning Attribute; Process; Protein Isoforms; prototype; Recurrence; Reporter; Research; response; Sensitivity and Specificity; small molecule; Specificity; Spectrum Analysis; Staging; Structure; Techniques; Testing; Therapeutic; Tissues; Tube; tumor; tumor progression; tumor xenograft; United States National Institutes of Health; Vertebral column; Whole Organism; Xenograft Model
Relevance: Narrative To develop near-infrared fluorescent beacons for the in vivo detection of phospholipase activity via dual optical imaging and MR spectroscopy
Project start date: 2009-05-01
Project end date: 2013-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01CA129176-03 (2011): $261450
MEDGAR EVERS/KINGSBOROUGH BRIDGE PROGRAM
J Edward, Professor
Medgar Evers Collegecity: New York country: United States (us)
Grant 5R25GM062003-10 from National Institute Of General Medical Sciences
Abstract: The Medgar Evers/Kingsborough Bridge Program will continue to upgrade an educational environment and mechanism to increase the number and quality of undergraduate students completing an Associate Degree in Science who continue to the Bachelor´s level with an ultimate aim of entering a career in biomedical research. The transition period between completing the A.S. degree and entering into a B.S. degree program can be a serious stumbling block, impeding the progress of well qualified and potentially successful students who continue to be grossly underrepresented in the fields of biomedical research. Our aim is to actively promote the academic success of students from the targeted areas by providing a five year long intervention and enrichment program using the combined resources of Medgar Evers College and Kingsborough Community College, both of which are located in Brooklyn, NY. The program will promote student interest, excitement and ability to pursue biomedical studies; student awareness of the accessibility of biomedical careers through continuation of their education; improve student verbal, written, mathematical and social skills to provide them with the basics to successfully pursue biomedical careers; promote student realization of the relationships of biomedical studies to all aspects of their lives because the biomedical sciences are a major driving force in society; and provide a smooth transition into a senior college for students graduating from Kingsborough Community College. To achieve this and better prepare students, who will be the next generation of biomedical researchers, Medgar Evers College and Kingsborough Community College will continue their academic partnership to improve the already good relationship, strengthen all meaningful components of our programs and initiate innovative strategies. The Medgar Evers/Kingsborough Bride Program has direct relevance to public health in so far as it will be a prime education conduit for a future generation of biomedical research scientists. The research training and projects that will be undertaken during the time period of the project are biomedically oriented. The students will be grounded in research training and go on the graduate schools and beyond to conduct biomedically relevant research
Keywords: Affect; Area; Awareness; Bachelor`s Degree; Biology; Biomedical Research; career; college; Communities; Development; Discipline; Doctor of Philosophy; driving force; Educational aspects; Educational Curriculum; Environment; Evaluation; experience; Faculty; Foundations; Future Generations; Goals; health disparity; Healthy People 2010; improved; innovation; Institution; Intention; interest; Internships; Intervention; Knowledge; meetings; Mission; next generation; programs; public health medicine (field); public health relevance; Qualifying; Research; Research Personnel; Research Project Grants; Research Training; Resources; Schools; Science; Scientist; social skills; Societies; Students; success; symposium; Time; United States Public Health Service; university student; Writing
Relevance: The Medgar Evers/Kingsborough Bride Program has direct relevance to public health in so far as it will be a prime education conduit for a future generation of biomedical research scientists. The research training and projects that will be undertaken during the time period of the project are biomedically oriented. The students will be grounded in research trainingand go on the graduate schools and beyond to conduct biomedically relevant research
Project start date: 2000-09-01
Project end date: 2014-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PAR-07-411
5R25GM062003-10 (2011): $207122
SCHISTOSOME EGG INDUCED TH2 RESPONSES
J Edward, Member
Trudeau Institute, Inc.city: Saranac Lake country: United States (us)
Grant 5R01AI032573-19 from National Institute Of Allergy And Infectious Diseases
Abstract: The helminth parasite Schistosoma mansoni is long lived, causing chronic infections in its natural human and experimental mouse hosts. It is recognized that during schistosomiasis the parasite-induced Th2 response, as measured by in vitro antigen-stimulated T cell proliferation or cytokine secretion assays, peaks early and then declines despite ongoing infection, a process that is referred to as immunomodulation. Immunomodulation in schistosomiasis appears to play a vital role in minimizing immunopathology in a setting where the immune system is incapable of eliminating the pathogen. Here we propose to explore the underlying basis and functional significance of the diminished Th2 responses that characterize chronic schistosomiasis. We hypothesize that chronic schistosome infection leads to a state of Th2 cell dysfunction akin to adaptive tolerance. We propose to test this hypothesis through three specific aims 1. To use IL-4 reporter mice to characterize Th2 cells throughout infection. 2. To establish whether Th2 cells from chronically infected mice are irreversibly dysfunctional or whether ongoing environmental signals are required to maintain them in this state. 3. To identify the mechanism responsible for immunomodulation during chronic infection. Recently, using 4get and KN2 IL-4 reporter mice, in which IL-4 transcription and protein production are reported by the expression of GFP and surface human CD2 respectively, we have been able to unequivocally identify CD4 T cells that have responded during infection and committed to Th2 differentiation by becoming capable of making IL-4. These cells can be detected immediately ex-vivo, and indeed in situ, and have allowed us to make significant steps forward in understanding what is happening during chronic schistosomiasis. Together, our studies promise to generate new insights into the processes that allow the regulation of Th2 responses during chronic schistosomiasis and are likely to be of relevance to other chronic helminth infections where Th2 cell hyporesponsiveness has been documented. We believe that our work has the potential to identify targets for intervention in important diseases of the developed world, such as asthma, allergy and ulcerative colitis, that are mediated by chronic, poorly controlled Th2 responses
Keywords: Acute; Antigens; Asthma; base; Biological Assay; Bromodeoxyuridine; CD4 Positive T Lymphocytes; Cell Count; Cell physiology; Cell Separation; Cells; Chronic; Commit; Coupled; cytokine; Dendritic Cells; Development; Disease; egg; Environment; falls; Functional disorder; Genetic Transcription; Granuloma; Helminthiasis; Helminths; Human; Hypersensitivity; Immune system; immunopathology; immunoregulation; In Situ; In Vitro; in vivo; Infection; insight; Interleukin-10; Interleukin-13; Interleukin-4; Intervention; Life; macrophage; Measures; Mediating; Molecular Profiling; Mus; Parasites; pathogen; Pathologic; Pharmaceutical Preparations; Phase; Physiologic pulse; Play; Process; Production; Proliferating; Property; Proteins; Regulation; Regulatory T-Lymphocyte; Reporter; Reporting; research study; Resistance; response; Role; Schistosoma; Schistosoma mansoni; Schistosomiasis; Signal Transduction; Staging; Surface; T-Cell Proliferation; Testing; Th2 Cells; Time; Tissues; Ulcerative Colitis; Work
Project start date: 1992-12-01
Project end date: 2013-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-07-070
5R01AI032573-19 (2011): $418770
STRUCTURE-FUNCTION RELATIONSHIPS OF PROTEIN 4.1 IN BLOOD
J Edward, Professor & President
Dana-farber Cancer Institutecity: Boston country: United States (us)
Grant 5R01HL044985-19 from National Heart, Lung, And Blood Institute
Abstract: Protein 4.1 R (4.1 R) was originally identified as an essential component of the red cell membrane cytoskeleton. We have shown that 4.1R plays a role in epithelial morphogenesis and endothelial cell-to-cell junction assemblies. We now propose to characterize the contributions of 4.1 R to tight junction (TJ) biogenesis and membrane polarity in epithelial cells, as well as to adherens junction (AJ) assembly and integrity in endothelial cells. The localization of 4.1 R during epithelial morphogenesis shifts from the nucleus and cytoplasm of non-confluent cells to the lateral membrane of confluent cells, in conjunction with a switch from 4.1R 135 kD, lacking exon 17b, to 4.1 R 160 kD containing exon 17b. In non-confluent nuclei, 4.1R associates with ZO-2. In confluent cells, 4.1R associates with the polarity protein Par3, TJs, and AJs. Dominant negative mutant 4.1R´s disrupt TJ integrity and produce an enlarged and flattened morphology. The behavior of 4.1 R during epithelial morphogenesis will be studied in both a model MDCK line and primary epithelial cultures. We shall dissect the mechanisms by which 4.1R associates with partner proteins to form polarized membranes and TJ´s using in vitro and in vivo biochemical assays, and fluorescence-based imaging methods. Functional perturbations will be achieved by utilizing siRNA, dominant-negative mutants, and a Tet-off expression system. In AJs, 4.1R co-localizes and associates with the AJ proteins E-cadherin and beta-catenin. Overexpression of 4.1 R reduces E-cadherin localization to the AJs. In order to study AJ´s in isolation, we shall use a well-characterized human umbilical vein endothelial cell (HUVEC) primary cell culture. Since HUVEC rarely express TJs, they are more suitable for analyzing the expression and sub-cellular distribution of 4.1 R and proteins with which it associates as they assemble into AJs. We shall delineate the impact of 4.1 R on AJs assembly, using approaches similar to those just described. We shall also test the hypothesis that complementarity may exist between 4.1R and its paralogues (4.1G, N, B), allowing them to compensate for the absence of 4.1 R in knock-out cells. Finally, we shall investigate the relationship between the shift in localization of 4.1 R from nucleus to peripheral membranes and the establishment of contact inhibition in terminally differentiating epithelial cells. These studies should yield new insights about the function of 4.1Rin cellular architecture and achievement of the post-mitotic contact inhibited state
Keywords: Achievement; Actins; Adherens Junction; Alternative Splicing; Antibodies; Architecture; base; Behavior; beta catenin; Biochemical; Biogenesis; Biological Assay; Biological Models; Blood; cadherin 5; Calcium; Cell division; cell growth regulation; Cell Maintenance; Cell membrane; Cell Nucleus; cell type; Cells; Complex; Contact Inhibition; Cytoplasm; Cytoskeleton; Defect; Dominant-Negative Mutation; E-Cadherin; Endothelial Cells; Epithelial; Epithelial Cells; Epithelium; Erythrocytes; Erythroid Cells; Exons; Fluorescence; Funding; gain of function; Glycophorin; Grant; Hemolytic Anemia; Human; imaging modality; In Vitro; in vivo; Inherited; insight; Intercellular Junctions; Knock-out; Knockout Mice; Lateral; Liquid substance; loss of function; MDCK cell; Membrane; membrane polarity; Membrane Proteins; Mitotic; Mitotic spindle; Mitotic Spindle Apparatus; Modeling; Monitor; Morphogenesis; Morphology; Movement; Nuclear; overexpression; Peripheral; Permeability; Phenotype; Play; Post-Translational Protein Processing; Primary Cell Cultures; Principal Investigator; Process; Promotor (Genetics); protein 4.1; Protein Analysis; protein complex; protein E; Protein Isoforms; Proteins; Role; shear stress; Signal Pathway; Site; Small Interfering RNA; Spectrin; Staging; Staining method; Stains; Structure-Activity Relationship; System; Techniques; Testing; Tetanus Helper Peptide; Tight Junctions; Tissues; Translation Initiation; Umbilical vein
Project start date: 1990-07-01
Project end date: 2011-07-31
Budget start date: 1-AUG-2009
Budget end date: 31-JUL-2011
5R01HL044985-19 (2009): $412345
5R01HL044985-18 (2008): $412345
Sponsored Links Excellgen http://Excellgen.com
5R01HL044985-17 (2007): $412345
2R01HL044985-16A1 (2006): $424250
NMR DETECTION OF TUMOR DIFFERENTIATION
J Edward, Research Associate Professor
University Of Pennsylvaniacity: Philadelphia country: United States (us)
Grant 5R01CA114347-05 from National Cancer Institute
Abstract: One of the most important goals that NCI has established in the area of cancer treatment has been for the conversion of cancer from an acute and lethal disease to a chronic condition that can be managed long term. Two requirements are needed to achieve this goal 1) the development of new anticancer drugs and treatment regimens utilizing these drugs, and 2) the ability to accurately and non-invasively assess the effectiveness of these treatment regimens. Differentiating agents are an attractive possibility in this area of cancer treatment, for they are agents that can either cause reversion of the malignant phenotype or the triggering of apoptosis. Our goal in this project is to delineate the mechanisms by which phenylacetate (PA) and phenylbutyrate (PB), two differentiating agents currently under Phase II clinical trials, inhibit the proliferation of cancer cells. We will concentrate on the changes in phosphatidylcholine (PtdCho) metabolites that can be observed using in vivo magnetic resonance spectroscopy (MRS) with the goal of determining the spectroscopic indicators that can reliably be used as an index for the inhibition of proliferation and induction of apoptosis by PA and PB. This could provide a basis for clinical monitoring of response to differentiation therapy by 1H MRS. In vivo animal tumor studies will be combined with mechanistic studies on perfused cells, using MR spectroscopy with fluorescence microscopy, flow cytometry and molecular biology techniques to identify key checkpoints in PtdCho catabolism associated with differentiation therapy. Our preliminary studies indicate that PA and PB induce changes in MR-visible phospholipid metabolite levels that correlate with the induction of apoptosis. Using fluorescent phospholipase-activated phospholipid analogues, we have identified two different phospholipase activities in prostate cancer cells one that is constitutive and primarily nuclear, and one that is cytoplasmic and inducible by PA or PB. Thus, the aims of this application are to i) measure in vitro and in vivo phospholipase activation induced by differentiating agents, ii) identify the phospholipase isoform contributing to spectral changes in PBinduced apoptosis; iii) examine the effects of phospholipase inhibition on the MR-visible metabolites in tumor cells undergoing apoptosis and iv) to investigate the potential of MR-visible resonances as a marker for response to differentiation therapy in murine tumor models. These studies provide a mechanistic underpinning for the changes induced in phospholipid metabolism by differentiation therapy and therefore constitute an important advance in the understanding of choline metabolism for the interpretation of in vivo MR spectra of tumors
Keywords: 2-Hydroxy-N, N, N-trimethylethanaminium; 3-hydroxy-3-methylglutaryl coenzyme A Inhibitors; 6-Methylcompactin; 85kDa Calcium-Independent Phospholipase A2; Acute; Acute Disease; acute disease/disorder; acute disorder; Agents, Cytostatic; analog; Animals; Anti-Cancer Agents; Anti-Tumor Agents; Anti-Tumor Drugs; anticancer agent; anticancer drug; anticancer therapy; Antineoplastic Agents; Antineoplastic Drugs; Antineoplastics; Antiproliferative Agents; Antiproliferative Drugs; Apopain; Apoptosis; Apoptosis Pathway; Apoptotic; Area; Assay; Attenuated; base; Bioassay; Biologic Assays; Biological; Biological Assay; Blood Coagulation Factor IV; Butanoic acid, 2-methyl-, 1, 2, 3, 7, 8, 8a-hexahydro-3, 7-dimethyl-8-(2-(tetrahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl)-1-naphthalenyl ester, (1S-(1alpha(R*), 3alpha, 7beta, 8beta(2S*, 4S*), 8abeta))-; Ca++ element; CaI-PLA(2); CaI-PLA2; Calcium; Calcium-Independent Cytosolic Phospholipase A2; Calcium-Independent Cytosolic Phospholipase A2 Group VI; Calcium-Independent Cytosolic Phospholipase A2VI; Calcium-Independent Phospholipase A2; cancer cell; Cancer Drug; Cancer of Prostate; cancer therapy; Cancer Treatment; Cancers; CASP-3; CASP3; Caspase 3, Apoptosis-Related Cysteine Protease; caspase-3; Catabolism; Cell Cycle; Cell Death; Cell Death, Programmed; Cell Division Cycle; Cell Growth in Number; Cell Line; Cell Lines, Strains; Cell Multiplication; Cell Proliferation; CellLine; Cells; Cellular Proliferation; Chemotherapeutic Agents, Neoplastic Disease; chemotherapy; Choline; Choline Glycerophospholipids; Choline Phosphoglycerides; Chronic; Chronic Disease; chronic disease/disorder; chronic disorder; Chronic Illness; Clinical; Clinical Trials, Phase II; Coagulation Factor IV; CPP-32; CPP32; CPP32 protein; CPP32B; CPP32beta; cultured cell line; Cysteine Protease CPP32; cysteine protease P32; Cytofluorometry, Flow; Cytosolic Phospholipase A2G6; Cytostatic Drugs; Cytostatics; cytotoxic; Death Rate; Detection; Development; Differentiating Agents; Differentiation Agents; Differentiation Inducer; Differentiation Therapy; Disease; disease/disorder; Disorder; Doctor of Philosophy; drug/agent; Drugs; DU145; EC 3.1.1.4; effective therapy; Effectiveness; Enzymes; Ethanaminium, 2-hydroxy-N, N, N-trimethyl-; Factor IV; fat metabolism; Flow Cytofluorometries; Flow Cytometry; flow cytophotometry; Flow Microfluorimetry; Fluorescence; Fluorescence Microscopy; fluorophore; G0 Phase; G0 state; Generations; Genital System, Male, Prostate; Goals; group VI phospholipase A(2); GVI PLA2; HMG-CoA Reductase Inhibitors; Human; human PLA2G6 protein; Human Prostate; Human Prostate Gland; Human, General; Hydroxymethylglutaryl CoenzymeA Reductase Inhibitors; Hydroxymethylglutaryl-CoA Reductase Inhibitors; In Vitro; in vivo; indexing; Induction of Apoptosis; Inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase; Inhibitors, Hydroxymethylglutaryl-CoA; Inhibitors, Hydroxymethylglutaryl-Coenzyme A; interest; Intermediary Metabolism; Investigators; iPLA(2); iPLA(2)-1; IPLA2; iPLA2 enzyme; Isoforms; Lecithin; lecithinase A; Lecithinase A2; Lecithinases; lipid metabolism; Lipids; Lovastatin; Magnetic Resonance Spectroscopy; malignancy; Malignant Cell; Malignant neoplasm of prostate; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; malignant phenotype; Malignant prostatic tumor; Malignant Tumor; Malignant Tumor of the Prostate; Mammals, Mice; Man (Taxonomy); Man, Modern; Measures; Medication; Metabolic; Metabolic Processes; Metabolism; Metabolism, Lipids/Lipoproteins/Membrane Constituents; METBL; Methods; Methods and Techniques; Methods, Other; mevacor; Mevinolin; Mice; Microfluorometry, Flow; Microscopy, Fluorescence; Microscopy, Light, Fluorescence; Modeling; Molecular; Molecular Biology Techniques; Monacolin K; Monitor; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; MR Spectroscopy; MRS; MRSI; Murine; Mus; National Cancer Institute; NCI; NCI Organization; necrocytosis; neoplasm/cancer; neoplastic cell; novel; Nuclear; One Step; One-Step dentin bonding system; PARP Cleavage Protease; Patients; Ph.D.; Pharmaceutic Preparations; Pharmaceutical Preparations; Phase 2 Clinical Trials; phase 2 study; phase 2 trial; Phase II Clinical Trials; phase II trial; PhD; Phenylacetates; Phenylbutyrates; phosphatidase; Phosphatides; phosphatidolipase; phosphatidylcholine 2 acylhydrolase; Phosphatidylcholines; Phospholipase; Phospholipase A2; Phospholipase A2 Group VI; Phospholipase A2VI; Phospholipid Metabolism; Phospholipids; Physiology; PLA2; PLA2G6; PLA2G6 protein, human; Production; programs; Programs (PT); Programs [Publication Type]; Proliferating; Prostate; Prostate CA; Prostate Cancer; Prostate Gland; Prostatic Cancer; Prostatic Gland; Protein Isoforms; protocol, phase II; Protocols, Treatment; QOL; Quality of life; Regimen; Research Personnel; Researchers; response; response marker; RGM; Role; SCA-1; social role; Solid Neoplasm; Solid Tumor; SREBP Cleavage Activity 1; Staging; Statins, HMG-CoA; study, phase II; System; System, LOINC Axis 4; Techniques; Therapeutic; Treatment Protocols; Treatment Regimen; Treatment Schedule; tumor; Tumor Cell; tumor xenograft; Tumor-Specific Treatment Agents; Xenograft Model; Yama; Yama protein
Project start date: 2005-05-13
Project end date: 2011-03-31
Budget start date: 1-APR-2009
Budget end date: 31-MAR-2011
5R01CA114347-05 (2009): $237454
MODELING BREAST CANCER RELAPSE PREVENTION IN MICE
J Edward, Assistant Professor
Pennsylvania State Univ Hershey Med Ctrcity: Hershey country: United States (us)
Grant 5R01CA152222-02 from National Cancer Institute
Abstract: Most breast cancer deaths result from relapse wherein sites of latent disease escape from a dormant state. Remarkably, the latent foci of residual disease that beget breast cancer relapse remain uncharacterized despite their enormous clinical importance. Therefore, very little is known about the biology of breast cancer dormancy and escape. Since sites of latent disease in breast cancer patients are likely to remain inaccessible to researchers for the foreseeable future, modeling dormant breast cancer in genetically-modified mice offers an alternative means to 1) uncover the biological basis for tumor dormancy, and 2) design and test rational treatment strategies aimed at eradicating dormant disease. With these goals in mind, a mouse mammary tumorigenesis model was developed in which a genetic event responsible for initiating breast cancer (activation of the Wnt signaling pathway) can be reversed. Shutting off Wnt signaling triggers regression of these cancers, but long-lived disease lesions persist and typically remain subclinical for extended periods before they "escape" from dormancy and beget tumor relapse. By recapitulating key clinical features of dormant breast cancer, this model provides novel experimental access to sites of latent disease, enabling elucidation of the cellular and molecular mechanisms that maintain dormancy. Moreover, the disease-free interval in this model provides a window for pre-clinical testing of treatment strategies directed against dormant cancer. In Aim1 of the proposal, conventional cytotoxic agents will be employed in pre-clinical modeling to determine how to optimally time treatment to prevent tumor escape. Here, outcomes will be compared when cytotoxic agents are administered before vs. during vs. after the initiation of dormancy. In Aim 2, genetically encoded reporters will be used to track tumor cell divisions within latent tumors to determine whether dormant mammary cancers are maintained by a subpopulation of quiescent, treatment-resistant tumor cells. These studies will test the concept that eradicating dormant cancer critically depends on targeting a quiescent subset of tumor cells that are crucial for relapse. In Aim 3, genetic pathways that drive escape from tumor dormancy will be discovered using a genetic screen that relies on the mobilization of a transposon ("jumping gene") in mammary cancers. This screen is designed to identify novel therapeutic targets for relapse prevention. Ultimately, our studies will aid in developing more rational approaches for preventing breast cancer relapse by eradicating sites of dormant disease. Most breast cancer deaths result from disease relapse in which sites of latent disease escape from a dormant state. Remarkably, almost nothing is known about the biology of tumor dormancy and escape. By modeling dormant breast cancer in genetically-modified mice, we propose to 1) uncover the biological basis for tumor dormancy, and 2) design and test rational treatment strategies aimed at eradicating dormant disease
Keywords: Address; After Care; After-Treatment; Aftercare; Antioncogene Protein p53; ARF tumor suppressor, mouse; base; Biological; biological signal transduction; Biology; Breast; Breast Adenocarcinoma; Breast Cancer Model; Breast Neoplasms; Breast Tumors; Cancer of Breast; Cancer Patient; cancer regression; Cancer Relapse; Cancers; Cdkn2a(p19ARF); Cell Communication and Signaling; Cell division; Cell Signaling; Cellular Tumor Antigen P53; Cessation of life; Clinical; Clinical Evaluation; clinical test; Clinical Testing; Cytotoxic agent; Cytotoxic drug; Cytotoxin; Death; design; designing; Detectable Residual Disease; Disease; Disease remission; Disease-Free Survival; disease/disorder; Disorder; disorder later incidence prevention; Event; Event-Free Survival; Future; Generalized Growth; Genetic; Genetic Alteration; Genetic Change; Genetic defect; Genetic Screening; Genetics-Mutagenesis; genome mutation; genome-wide; Goals; Growth; Human; Human, General; Immune Escape, Tumor; intervention therapy; Intracellular Communication and Signaling; Investigators; Jumping Genes; Knowledge; Label; Lesion; Life; malignancy; Malignant; Malignant - descriptor; malignant breast neoplasm; Malignant neoplasm of breast; Malignant Neoplasms; Malignant Tumor; Malignant Tumor of the Breast; Mammals, Mice; mammary; Mammary adenocarcinoma; Mammary Cancer; mammary cancer model; Mammary gland; Mammary Glands, Human; Mammary Neoplasms; mammary tumor; mammary tumor model; Mammary Tumorigenesis; Man (Taxonomy); Man, Modern; Mediating; Mice; Mind; Minimal Residual Disease; Modeling; Molecular; Molecular Biology, Mutagenesis; Monitor; mouse model; Murine; Mus; Mutagenesis; mutant; Mutation; neoplasm recurrence; Neoplasm, Residual; neoplasm/cancer; neoplastic cell; new therapeutic target; novel; Oncogenic; Oncoprotein p53; ontogeny; Outcome; p19(ARF); p19(ARF) protein; p19(ARF) protein, mouse; p19ARF; p53 Antigen; p53 Tumor Suppressor; pathway; Pathway interactions; Patients; Phosphoprotein P53; Phosphoprotein pp53; Physiologic pulse; Population; Pre-Clinical Model; Preclinical Models; prevent; preventing; prevention of disease recurrence; prevention of disorder recurrence; prevention of later incidences of a disorder; prevention of recurrence; Prevention of relapse; Property; Property, LOINC Axis 2; Protein p53; Protein TP53; Protocols, Treatment; public health relevance; Publishing; Pulse; reconstitute; reconstitution; Recurrent disease; Recurrent Neoplasm; Recurrent tumor; Regimen; Relapse; Relapsed Disease; Remission; Reporter; Reporting; research clinical testing; Research Personnel; Researchers; residual disease; Residual Neoplasm; Residual Tumors; Resistance; resistant; response; RGM; RMSN; Role; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Simulate; Site; Sleeping Beauty; social role; Testing; Therapeutic Intervention; Time; Tissue Growth; TP53; Treatment Protocols; Treatment Regimen; Treatment Schedule; treatment strategy; tumor; Tumor Biology; Tumor Cell; Tumor Escape; Tumor Protein p53; Tumors, Residual; Work
Relevance: Most breast cancer deaths result from disease relapse in which sites of latent disease escape from a dormant state. Remarkably, almost nothing is known about the biology of tumor dormancy and escape. By modeling dormant breast cancer in genetically-modified mice, we propose to: 1) uncover the biological basis for tumor dormancy, and 2) design and test rational treatment strategies aimed at eradicating dormant disease
Project start date: 2010-08-18
Project end date: 2015-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01CA152222-02 (2011): $322272
MOLECULAR GENETIC TOOLS FOR PARASITIC HELMINTHS
J Edward, Professor
University Of Pennsylvaniacity: Philadelphia country: United States (us)
Grant 5R01AI082548-03 from National Institute Of Allergy And Infectious Diseases
Abstract: Helminth (worm) parasites take an enormous toll on human health, especially in developing countries. Half a billion people suffer debilitating, sometimes fatal illness as a result of these infections, and subclinical effects of helminth parasitism include retarded physical and cognitive development. Despite their global health impact, and the pressing need for new treatments and preventative measures against them, there has been a decline in the numbers of investigators funded to work on these pathogens, and a consequent reduction in the number of young scientists choosing to enter this field. The paucity of modern molecular tools for interrogating gene function in parasitic helminths and the resulting degree to which helminth biology remains descriptive as opposed to mechanistic in nature are undoubtedly factors in this decline. Nevertheless, the free-living worm Caenorhabditis elegans and planaria can be studied using sophisticated molecular tools, suggesting that development of similar tools for parasitic worms is feasible. In this vein, we and others have had recent encouraging success in the experimental manipulation of gene expression in parasitic worms. Despite this, significant technical hurdles remain if molecular genetics is to become routine in helminth parasitology. Therefore, we propose to further develop and refine tools for the manipulation of gene expression in helminth parasites. We will focus on two important human pathogens, Strongyloides stercoralis and Schistosoma mansoni. These represent, respectively, the Nematoda and Platyhelminthes, two phyla of medically important helminths. The first of our two specific aims is to develop transgenic parasitic helminths that are amenable to experimentation. Work towards this aim will involve developing DNA constructs that allow regulated, tissue-specific transgene expression in transiently transformed S. mansoni and St. stercoralis. Also, we will seek to identify regulatory sequences that allow conditional transgene expression and both intra- and extracellular transport of recombinant proteins. We will also develop methods for establishing stably expressing transgenic lines through serial host passage. New methods for chromosomal integration of transgenes will be developed and insulator sequences will be identified as measures against epigenetic transgene silencing in both transiently and stably transformed worms. Under our second specific aim we will target expressed genes for silencing in S. mansoni and St. stercoralis. Hypotheses that the activities of RNAi processing enzymes homologous to C. elegans RDE-1, SAGO-1, SID-1 and SID-2 limit the silencing efficiency of exogenously applied dsRNA in St. stercoralis and that siRNAs administered to the host can silence genes of S. mansoni and St. stercoralis in vivo are to be tested under this aim. The proposed research has the potential to create new tools that will invigorate the molecular and cellular biological study of a neglected and important group of human pathogens. This application proposes to develop new methods to study gene function at the molecular level in Schistosoma mansoni and Strongyloides stercoralis, representatives of two major groups of parasitic helminths (worms). Its two specific aims propose to develop methods for DNA transformation of S. mansoni and St. stercoralis with transgenes capable of regulated tissue specific expression and of stable expression through serial host passage, and for targeted silencing or disruption of gene expression in the two parasites. Because reliable techniques of this nature are either underdeveloped or altogether unavailable, the proposed project has the potential to invigorate the field of parasitic helminthology, which is currently perceived as largely descriptive, as opposed to mechanistic, in nature
Keywords: Biological; Biology; Caenorhabditis elegans; Childhood; Cognitive; Communities; Country; Development; DNA; Double-Stranded RNA; Enzymes; Epigenetic Process; extracellular; Funding; Gene Expression; gene function; Gene Silencing; Gene Transfer Techniques; Genes; global health; Goals; Health; Helminths; Human; in vivo; Infection; Intervention; Knowledge; Life; Measures; Medical; Methodology; Methods; Molecular; Molecular Genetics; Nature; neglect; Nematoda; new technology; Parasites; Parasitic nematode; parasitism; Parasitology; pathogen; Platyhelminths; Process; public health medicine (field); Recombinant Proteins; Research; Research Personnel; Resistance; RNA Interference; Schistosoma mansoni; Science; Scientist; Strongyloides stercoralis; success; Techniques; Testing; Tissues; tool; transgene expression; Transgenes; Transgenic Organisms; Vaccines; Veins; Work
Project start date: 2009-05-01
Project end date: 2014-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: RFA-AI-08-005
5R01AI082548-03 (2011): $350831
J Edward, Professor & President
Dana-farber Cancer Institutecity: Boston country: United States (us)
Grant 3P30CA006516-46S6 from National Cancer Institute
Abstract: Dana Farber/Harvard Cancer Center (DF/HCC) is an inter-institutional research enterprise that unites the major clinical, population, and basic cancer research efforts of the Harvard medical and public health community. It was founded in 1998 through a formal agreement of the Dana-Farber Cancer Institute, the Harvard Medical School and Harvard School of Medicine and Public Health, and the Beth Israel Deaconess, Brigham and Women´s, Children´s, and Massachusetts General Hospitals. By agreement, this will be the only NCI-designated Cancer Center application from these institutions. The DF/HCC links the efforts of a large cadre of cancer scientists - currently more than 800 members with NCI grants amounting to more than $200 million in total costs-in an interlocking organizational structure designed to promote research advances that are aimed at lowering the burden of cancer. In the aggregate, Center members have extensive experience and well-recognized expertise in the three major cancer research disciplines basic, clinical, and population science. The research of the Center is carried out in 17 disease site- and discipline-based Research Programs that cross both institutional and scientific boundaries. In addition, the Center supports 19 Core Facilities that make shared resources, dedicated to enhancing scientific efficiency, available to all Center members. The primary goal of the Center is to promote collaborative interactions that will lead to new approaches to cancer prevention, diagnosis, and treatment
Keywords: Agreement; anticancer research; base; Basic Cancer Research; Basic Science; Cancer Burden; Cancer Center; cancer prevention; Child; Clinical; Clinical Sciences; Communities; Core Facility; cost; Dana-Farber Cancer Institute; design; Diagnosis; Discipline; Disorder by Site; experience; General Hospitals; Goals; Grant; Institution; Israel; Lead; Link; Malignant Neoplasms; Massachusetts; Medical; medical schools; member; NCI-Designated Cancer Center; novel strategies; organizational structure; Population; Population Sciences; programs; public health medicine (field); Research; Resource Sharing; Scientist; Woman
Project start date: 1977-12-01
Project end date: 2011-11-30
Budget start date: 1-DEC-2009
Budget end date: 30-NOV-2011
3P30CA006516-46S6 (2011): $75000
5P30CA006516-46 (2010): $11641774
3P30CA006516-46S1 (2010): $124531
3P30CA006516-46S2 (2010): $100000
3P30CA006516-46S3 (2010): $100000
Sponsored Links Excellgen http://Excellgen.com
3P30CA006516-46S4 (2010): $46458
3P30CA006516-46S5 (2011): $5876096
BIOSTATISTICS AND BIOINFORMATICS SHARED RESOURCE
J Edward, Professor
University Of New Mexicocity: Albuquerque country: United States (us)
Keywords: Archives; base; Bioinformatics; Bioinformatics Shared Resource; Biomedical Research; Biometry; Biostatistical Methods; Cancer Center; Clinical; Clinical Research; Clinical Trials; cohort; Collaborations; Communities; Computational Biology; computer science; Computer software; Data; Data Element; data exchange; data portal; Data Set; Databases; design; Development; Disease; Engineering; Epidemiologic Studies; Faculty; Fostering; Funding; Generations; Genomics; Human Resources; Instruction; Internet; Knowledge; Laboratory Study; Lead; Malignant Neoplasms; Mathematics; meetings; member; Methods; Modeling; New Mexico; novel; Patients; Pattern; programs; repository; Research; Research Design; Research Personnel; Research Project Grants; Resources; Schedule; Science; Scientist; Services; statistics; Technology; Training; Universities; web site
Relevance: RELEVANCE (See instructions): The Resource enhances the clinical trials and Research Programs of the Cancer Center by providing critical expertise in the design and analysis of cancer related studies. These research efforts would be not Just hampered, but often rendered meaningless, without appropriate biostatistical and bioinformatics support. The Resource also provides statistical support during the development of projects that will ultimately lead to both increased research funding to the Cancer Center and support for members of the Resource. Furthermore, the Bioinformatics technical staff members provide the mechanisms by which large-scale datasets and associated clinical covariate data generated by Cancer Center researchers are reliably managed, stored, annotated, archived, and disseminated. These activifies are critical to meeting the Specific Aims of Cancer Center research grants that are based on the generation and analysis of large-scale genomic datasets for large patient cohorts and emerging large-scale deep sequencing data
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
5P30CA118100-07_6325 (2011): $88880