UNIVERSITY OF SOUTHERN CALIFORNIA (USC)
V David, Assistant Professor
University Of Pennsylvaniacity: Philadelphia country: United States (us)
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5U01DA020830-07_8127 (2011): $16592
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to V David
DETECTION AND DIAGNOSIS OF PANCREATIC CARCINOMA
V David, Professor
Ctr For Molecular Medicine/immunologycity: Belleville country: United States (us)
Grant 5R01CA096924-06 from National Cancer Institute
Abstract: The objective of this research proposal is the development and evaluation of immunoassays having clinical utility for the "early" detection and diagnosis of pancreatic carcinoma. In the year 2009, an estimated 42,000 new cases of pancreatic carcinoma will be diagnosed in the United States. Pancreatic carcinoma is the tenth most common form of cancer in men and women today, yet it is the fourth leading cause of cancer deaths. The most common symptoms of the disease, jaundice, abdominal pain, and weight loss, together with other presenting factors are nonspecific in nature. Thus, diagnosing pancreatic carcinoma at an early stage of tumor growth is difficult at best, requiring considerable suspicion and extensive diagnostic work-up, up to and including exploratory surgery. Our laboratory has developed and characterized the PAM4 monoclonal antibody (murine, chimeric and humanized versions), providing evidence as to its potential for clinical detection, imaging and therapy of pancreatic carcinoma. Our recent developments employing an in vitro enzyme immunoassay to quantitate PAM4-reactive antigen in the blood of patients, appears quite promising for detection of pancreatic carcinoma and its discrimination from pancreatitis, as well as other cancers and normal individuals. Early detection and diagnosis of pancreatic carcinoma, as well as appropriate staging of the disease, would almost certainly provide a survival advantage. With this in mind, we intend to further develop and assess the ability of the PAM4-based immunoassays, both immunohistochemical detection in tissue specimens and enzyme immunoassay to detect and quantitate the antigen in both serum and pancreatic fluids, to provide "early" detection and diagnosis at a time-point when therapeutic intervention may have better opportunity for successful outcome. The specific aims of this project are 1. To evaluate the performance profile of the PAM4-Immunoassay for diagnostic accuracy; 2. To evaluate the clinical utility of the PAM4-Immunoassay for a-accurate diagnosis of patients presenting with symptoms suggestive of pancreatic cancer; b- early detection of pancreatic carcinoma in asymptomatic individuals at high risk for pancreatic cancer; c- assessment of tumor response or early detection of relapse; 3. To evaluate the nature of the antigen to which PAM4 is reactive. Pancreatic carcinoma is an insidious disease with a particularly high mortality rate. In large measure, this is due to the location of the tumor where it can grow in a silent fashion. Symptoms that might suggest the patient seek medical assistance are usually not evident until an advanced stage of tumor growth. Compounding this challenge is that currently available treatment procedures have not been able to provide a cure for the overwhelming majority of patients. Our goal is to provide an antibody approach for early detection and diagnosis of the disease at a time when curative procedures have a better opportunity for successful outcome. We have developed monoclonal antibody PAM4 that has a high specificity for pancreatic carcinoma as compared to benign pancreatic disease, other types of malignancies, and normal individuals. Thus, the detection of the PAM4-reactive antigen provides a high likelihood for the diagnosis of pancreatic cancer. The aims of the proposed research are to further define the specificity of the antibody as to its ability to provide accurate diagnoses, and to examine the value of the immunoassay for "early" detection of pancreatic carcinoma, prior to the development of symptoms, in a group of patients considered to be at high risk for development of the disease
Keywords: Abdominal Pain; Affinity; Antibodies; Antibody Specificity; Antigens; base; Benign; Biochemical; Biological Assay; biomarker; Blood; Body Weight decreased; cancer diagnosis; Cancer Etiology; Carcinoma; cell type; Cessation of life; Clinical; Clinical Trials; Data; Data Correlations; Detection; Development; Diagnosis; Diagnostic; diagnostic accuracy; Discrimination (Psychology); Disease; disease diagnosis; Disease Progression; Early Diagnosis; Enzyme Immunoassay; Epitopes; Evaluation; follow-up; Funding; Generations; Glycoproteins; Goals; Grant; Hepatobiliary; high risk; Histocompatibility Testing; Histologic; Icterus; Image; Immunoassay; improved; In Vitro; Individual; Investigation; Laboratories; Lesion; Liquid substance; Location; Malignant - descriptor; Malignant neoplasm of pancreas; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Medical Assistance; men; Mind; Modification; Monoclonal Antibodies; Mortality Vital Statistics; Mucins; Mus; Nature; Normal tissue morphology; Observational Study; Operative Surgical Procedures; Outcome; outcome forecast; Pancreas; Pancreatic carcinoma; Pancreatic Diseases; Pancreatic Intraepithelial Neoplasia; pancreatic neoplasm; Pancreatitis; Patients; Peptides; Performance; Phage Display; Population; Predictive Value; Procedures; programs; Property; prospective; public health relevance; Publications; Publishing; Relapse; Reporting; Research; Research Proposals; response; Role; Sampling; Screening procedure; Serum; Specificity; Specimen; Staging; statistics; Study Section; Surveillance Program; Symptoms; Systemic Therapy; Testing; Therapeutic Intervention; Time; Tissues; Translating; tumor; tumor growth; United States; Woman; Work
Project start date: 2002-07-01
Project end date: 2015-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-070
5R01CA096924-06 (2011): $252372
HIGH-THROUGHPUT PLATFORM FOR IDENTIFYING STEM CELL TOXICITY
V David
Rensselaer Polytechnic Institutecity: Troy country: United States (us)
Grant 1R01ES020903-01 from National Institute Of Environmental Health Sciences
Abstract: The characterization and safety assessment of drugs, drug candidates, environmental chemicals, etc. requires extensive resources, particularly when animal studies are employed. Thus, there is a need to develop new in vitro techniques to predict which compounds pose an increased threat to human health and should therefore be prioritized for further screening and evaluation. Because of the critical role self-renewing stem cells play in tissue function and homeostasis, retention of healthy adult stem cells is crucial for human health. Nevertheless, the different responses of human stem cells compared with terminally differentiated cell types (e.g., primary and transformed cell lines) against drugs, drug candidates, and chemicals have been only sparsely studied. Such information would be vital to identify the toxic effects of such chemicals on the human body and in specific organs. We hypothesize that stem cells may behave in a fundamentally different way from the differentiated cells often used for toxicity studies, and we will address this hypothesis through the use of appropriate model systems. An outcome of this study, then, is that toxicity studies must be expanded to include stem cells, or drugs will proceed into trials with missing and even misleading toxicity information. To test this hypothesis, we propose to develop a high-throughput, microscale 3D cell-based screening tool that will enable rapid and highly quantitative information to be obtained on the effects of drugs, drug candidates, and chemical toxicants on human stem cells; specifically neural stem cells (NSCs) in comparison to adult primary and transformed neural cells, and human mesenchymal stem cells (MSCs). The specific aims of the proposed work are to 1. Establish a robust 3D cell culture chip platform to grow and differentiate NSCs and MSCs; 2. Perform combinatorial in vitro extracellular matrix development; 3. Develop high-throughput on-chip assays of key signaling pathways in NSCs and MSCs that are critical in proliferation (self-renewal) and differentiation; 4. Validate the 3D cell culture microarray platform for cytotoxicity of stem cells in comparison to terminally differentiated primary cells and transformed cell lines using a series of reference compounds. This information is essential for the development of high throughput predictive toxicology screens for drug discovery and the prioritization of environmental chemicals based on their potential human toxicity. The proposed effort impacts human health by understanding the critical differences that chemical toxicants (e.g., drugs, environmental chemicals, etc.) have on human adult stem cells in relation to non-stem cells. A wealth of information on specific responses of different human adult stem cells will be obtained as a result of exposure to various known chemical toxicants. In addition, a new high-throughput screening tool will be developed for predictive human toxicology. This work will facilitate the use of stem cells in environmental health screening and in early-stage drug toxicity screening because of the critical role self-renewing stem cells play in the health of an individual
Keywords: Address; Adult; adult stem cell; Animals; Astrocytes; Astrocytoma; base; Biochemical; biochip; Biological Assay; Biological Models; Biotechnology; cell behavior; Cell Culture System; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell Survival; cell type; Cells; Chemical Engineering; Chemicals; combinatorial; cytotoxicity; Development; drug candidate; drug discovery; Drug toxicity; Engineering; Environment; environmental chemical; Environmental Health; Evaluation; Exposure to; Extracellular Matrix; Glial Fibrillary Acidic Protein; Goals; Growth; Health; high throughput screening; Homeostasis; Human; human adult stem cell; Human body; human stem cells; Immunofluorescence Immunologic; In Vitro; in vivo; Individual; MAP Kinase Gene; Mesenchymal Stem Cells; Methods; Nature; nerve stem cell; nestin protein; Neurons; Organ; Osteocalcin; Outcome; Outcome Study; Pathway interactions; Performance; Pharmaceutical Preparations; Phase; Play; Poisons; Property; Research; Resources; response; Role; Safety; Screening procedure; self-renewal; Series; Signal Pathway; Staging; stem cell biology; stem cell differentiation; stem cell fate; stem cell niche; Stem cells; Techniques; Technology; Testing; Tissues; tool; Toxic effect; toxicant; Toxicology; Transformed Cell Line; Transport Process; Work
Relevance: The proposed effort impacts human health by understanding the critical differences that chemical toxicants (e.g., drugs, environmental chemicals, etc.) have on human adult stem cells in relation to non-stem cells. A wealth of information on specific responses of different human adult stem cells will be obtained as a result of exposure to various known chemical toxicants. In addition, a new high-throughput screening tool will be developed for predictive human toxicology. This work will facilitate the use of stem cells in environmental health screening and in early-stage drug toxicity screening because of the critical role self-renewing stem cells play in the health of an individual
Project start date: 2011-12-23
Project end date: 2015-10-31
Budget start date: 23-DEC-2011
Budget end date: 31-OCT-2012
1R01ES020903-01 (2012): $532771
DIABETOGENIC ROLE OF MHC CLASS I ALLELES IN NOD MICE
V David, Associate Staff Scientist
Jackson Laboratorycity: Bar Harbor country: United States (us)
Grant 5R01DK046266-17 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Multiple susceptibility (Idd) genes contribute to T cell mediated autoimmune type 1 diabetes (T1D) in both humans and NOD mice, but with particular MHC haplotypes providing the primary risk factor. While also present in many non-autoimmune prone strains, the H2g7 MHC haplotype encoded Kd and/or Db class I molecules play an essential diabetogenic role when expressed in NOD mice. The overall goal of this project continues to be determination of the immunogenetic basis for the aberrant generation of diabetogenic MHC class I dependent CD8 T cells in NOD mice, with the hope such knowledge could ultimately provide translational information for blocking development of similar effectors in humans at high future disease risk. In NOD mice the H2g7 class I gene products aberrantly lose ability to mediate the intrathymic negative selection of diabetogenic CD8 T cells. A currently unknown polymorphic gene(s) within the Idd7 locus plays a major interactive role in determining whether H2g7 class I restricted diabetogenic CD8 T cells undergo negative selection. Hence, specific aim 1 is to more finely map, functionally characterize, and potentially identify the Idd7 region gene(s) regulating the negative selection efficiency of diabetogenic CD8 T cells. There is now also compelling evidence in humans that when expressed in the right genetic context, certain quite common MHC class I variants, such as HLA-A2.1, aberrantly contribute to T1D development. Supporting this situation is our finding that NOD. ¿2mnull.HHD mice expressing human HLA-A2.1, but no murine MHC class I molecules, continue to develop T1D. This was not a generic function of any human class I variant transgenically expressed in NOD mice. Previous work identified a series of pancreatic ¿ cell derived peptides that are presented by HLA-A2.1 class I molecules to diabetogenic CD8 T cells in NOD. ¿2mnull.HHD mice. T1D protective tolerance can be induced to pancreatic ¿ cell peptides that are presented by H2g7 class I molecules in standard NOD mice through appropriated dosed injections into young pre-disease state recipients. Diabetogenic CD8 T cells in standard NOD mice also undergo negative selection if forced to mature in the presence of engrafted antigen presenting cells (APC) expressing some non-H2g7 MHC haplotypes. Specific aim 2 will determine if diabetogenic HLA-A2.1 restricted T cell responses in NOD. ¿2mnull.HHD mice can be attenuated by a potentially tolerogenic peptide administration or APC engraftment protocol. In the currently available NOD. ¿2mnull.HHD stock only murine T cell receptor (TCR) molecules can engage antigens presented by the human HLA-A2.1 class I variant. Because of their respective human or murine origin, the TCR molecules expressed by diabetogenic HLA-A2.1 restricted T cells in T1D patients and NOD.¿2mnull.HHD mice might vary in their antigen binding affinities, which in turn could result in differential selection and/or activation effects. Thus, specific aim 3 is to functionally characterize NOD.22mnull.HHD mice transgenically expressing a human T1D patient derived TCR that recognizes a known HLA-A2.1 restricted pancreatic ss cell autoantigen. Type 1 diabetes (T1D) is a life threatening disease that results from an aberrant T lymphocyte mediated autoimmune response resulting in destruction of insulin producing cells in the pancreas. The goal of this current renewal application is to determine, through a series of mouse models, how a particular population of autoreactive T lymphocytes contributing to T1D aberrantly develop, and identify means that can stop this process. Such information may ultimately identify means that can attenuate the development of similar autoreactive T lymphocytes contributing to T1D in human patients
Keywords: accessory cell; Address; Affinity; Alleles; Allelomorphs; antigen binding; Antigen-Presenting Cells; Antigens; APC; ATGN; Attenuated; Autoantigens; Autoimmune; Autoimmune Process; Autoimmune Responses; Autologous Antigens; base; Caucasian; Caucasian Race; Caucasians; Caucasoid; Caucasoid Race; CD4 lymphocyte; CD4 Positive T Lymphocytes; CD4 T cells; CD4+ T cell; CD4+ T-Lymphocyte; CD4-Positive Lymphocytes; CD8; CD8B; CD8B1; CD8B1 gene; Cells; Cells, CD4; design; designing; Development; Diabetes Mellitus, Brittle; Diabetes Mellitus, Insulin-Dependent; Diabetes Mellitus, Juvenile-Onset; Diabetes Mellitus, Ketosis-Prone; Diabetes Mellitus, Sudden-Onset; Diabetes Mellitus, Type 1; Diabetes Mellitus, Type I; Disease; disease risk; disease/disorder; Disorder; disorder risk; Dose; Drugs, Nonproprietary; Engraftment; Future; Gastrointestinal Tract, Pancreas; gene product; Generations; generic; Generic Drugs; Genes; Genes, Class I; Genes, MHC Class I; Genetic; Goals; Haplotypes; Health; helper T cell; Histocompatibility Complex; Histocompatibility Complices; HLA-A2.1; Human; Human, General; Humulin R; IDD; IDDM; immunogen; Immunogenetic; Immunogenetics; Immunologic Accessory Cells; Inbred NOD Mice; Injection of therapeutic agent; Injections; Insulin; Insulin (ox), 8A-L-threonine-10A-L-isoleucine-30B-L-threonine-; insulin dependent diabetes; Insulin, Regular; Insulin-Dependent Diabetes Mellitus; Investigators; juvenile diabetes; juvenile diabetes mellitus; ketosis prone diabetes; Knowledge; Life; LYT3; Major Histocompatibility Complex; Major Histocompatibility Complex Receptor; Major Histocompatibility Complices; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Mediating; MHC Class I; MHC Class I Genes; MHC Receptor; Mice; Mice, Inbred NOD; Monocytes / Macrophages / APC; mouse model; Mouse, NOD; Murine; Mus; Non-Obese Diabetic Mice; Nonobese Diabetic Mouse; Novolin R; Occidental; Pancreas; Pancreatic; Patients; Peptides; Play; Population; Predisposition; Process; Proteins; Protocol; Protocols documentation; Public Health; public health medicine (field); Receptors, Antigen, T-Cell; Research Personnel; Researchers; Resistance; resistant; response; Risk Factors; Role; Self-Antigens; Series; social role; Susceptibility; T cell response; T-Cell Development; T-Cell Ontogeny; T-Cell Receptor; T-Cells; T-Lymphocyte; T-Lymphocyte Development; T1 diabetes; T1D; T1DM; T4 Cells; T4 Lymphocytes; thymus derived lymphocyte; Thymus-Dependent Lymphocytes; Time; Type 1 diabetes; type I diabetes; Variant; Variation; white race; Work
Relevance: Relevance to Public Health Type 1 diabetes (T1D) is a life threatening disease that results from an aberrant T lymphocyte mediated autoimmune response resulting in destruction of insulin producing cells in the pancreas. The goal of this current renewal application is to determine, through a series of mouse models, how a particular population of autoreactive T lymphocytes contributing to T1D aberrantly develop, and identify means that can stop this process. Such information may ultimately identify means that can attenuate the development of similar autoreactive T lymphocytes contributing to T1D in human patients
Project start date: 1993-06-01
Project end date: 2014-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01DK046266-17 (2011): $338016
LOCALIZING OBJECTS IN DYNAMIC SCENES
V David, Associate Professor
University Of California Berkeleycity: Berkeley country: United States (us)
Grant 5R01EY018216-05 from National Eye Institute
Abstract: Most visual and visuomotor functions hinge on our visual system´s ability to localize objects in dynamic scenes-to determine the positions of objects as we move or as objects in the world move around us. However, dynamic localization requires that the visual system must be able to both assign and update object positions, and it remains unclear which visual areas play these necessary roles and what mechanisms are responsible. Assigning and updating object locations is especially relevant considering that the eye is rarely stabilized and objects are constantly drifting across the retina as they move. Given the sluggish nature of visual processing, how do we perceive the positions of moving objects with clarity and precision? To understand how the visual system assigns and updates object positions, we must approach the task of localization not as a static process, but as a dynamic one. The goal of our proposed experiments is to characterize the neural pathways and mechanisms that determine perceived position in dynamic situations. Our preliminary results suggest that two mechanisms are required to perceive the positions of objects when motion is present on the retina trailing edge deblurring and edge-shifting. Without these mechanisms, we would perceive the world as a blurry smear, unable to accurately predict the future locations of objects or interact with them as they move around us. We hypothesize that these two independent mechanisms integrate information between early visual areas (V1) and the motion sensitive region (MT+). Our pilot data for the proposed experiments suggest that (1) MT+ carries precise information about object position, supporting the hypothesis that it serves an integral role in object localization, and (2) feedback as well as feed-forward connections between V1 and MT+ may be necessary to assign and update the positions of objects when retinal motion is present. Using functional magnetic resonance imaging (fMRI), transcranial magnetic stimulation (TMS), and psychophysics, we will characterize and isolate the neural locus of the two mechanisms and establish their causal roles in perceiving object position. Dynamic localization is the default situation faced by the visual system, but many neurological disorders produce specific deficits in dynamic visual and visuomotor localization, including optic ataxia, dyslexia, akinetopsia, autism, and schizophrenia, among others. Until we understand how position is determined in the normal brain for images that are constantly moving across the retina, we lack the necessary insight to develop diagnostic tools, predictive markers, and therapies for these impairments
Keywords: area V1; Ataxia; Autistic Disorder; base; Brain imaging; Code; Data; Dependence; Diagnostic; Dyslexia; Environment; extrastriate visual cortex; Eye; Feedback; feeding; Functional Magnetic Resonance Imaging; Future; Goals; Impairment; insight; Link; Location; Measures; Modeling; Motion; Nature; nervous system disorder; Neural Pathways; neuromechanism; Optics; Pattern; Play; Positioning Attribute; Process; programs; Property; Psychophysics; Psychophysiology; relating to nervous system; Research Personnel; research study; Resolution; response; Retina; Retinal; retinotopic; Role; Schizophrenia; Testing; Therapeutic; tool; Transcranial magnetic stimulation; Update; Visual; Visual Motion; visual motor; visual process; visual processing; Visual system structure
Project start date: 2007-04-01
Project end date: 2012-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5R01EY018216-05 (2011): $336470