HARNESSING B LYMPHOCYTES AS ANTIGEN-SPECIFIC REGULATORS OF ISLET TOLERANCE
J Daniel
Vanderbilt Universitycity: Nashville country: United States (us)
Grant 1K08DK090146-01 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: The PI seeks to develop a career as a physician-scientist focused on the problem of restoring and maintaining immune tolerance in individuals with Type 1 diabetes (T1D) by extending his scientific training in the exceptional environment at Vanderbilt University Medical Center. The failure of immune tolerance is an important underpinning of all human autoimmune diseases. When tolerance fails, autoimmune processes damage vital organs in tens of millions of patients worldwide. will focus on this scientific challenge through fundamental investigation of the most common pediatric autoimmune disorder-T1D, which afflicts more than 2 million Americans. To prevent and reverse this disease, it is necessary to restore immune tolerance to islet antigens. Most present efforts focus on either limiting the activation of islet-destructive T lymphocytes or enhancing the capacity of certain islet-protective T cells for immune regulation. Significantly, B lymphocytes also play a key role in the disease process both through the production of autoantibodies, which predict disease risk, and by the action of B lymphocytes as requisite antigen presenting cells. Targeting these cells with the B cell depleting agent rituximab has been at least as effective as T cell targeted agents, such as anti-CD3, in slowing the progression of new-onset diabetes. Moreover, studies in a relevant murine model of human T1D suggest a role of regulatory B cells in this salutary effect. These regulatory functions of B cells have also been observed in other models of autoimmune disease, and has previously characterized their role in transplantation tolerance. Building on these seminal data, we hypothesize that B lymphocytes play a regulatory role in the establishment and maintenance of peripheral immune tolerance and that this capacity is disrupted in T1D. This proposal investigates testable hypotheses which will focus on defining the regulatory functions of B lymphocytes that contribute to islet tolerance by determining the role of classical B cell subsets, antigen specificity and developmental signals in regulatory B cell function (Aim 1), and their cellular targets and the mechanisms through which their regulatory function is controlled (Aim 2). The environment at Vanderbilt University is poised to address this fundamental biologic question given the local expertise in B lymphocyte immunobiology, the exceptional training atmosphere created by the Vanderbilt Diabetes Center, and the dedication to physician-scientist development exemplified by the Departments of Pediatrics and Microbiology and Immunology. Investigation of these novel hypotheses under the guidance of an exceptional mentoring committee and supported by these outstanding resources will allow the PI to realize a significant opportunity for new discovery in this clinically relevant field of inquiry-the fundamental problem of lost tolerance in T1D. Type 1 diabetes is an unavoidable, life-long illness with no known cure and an imperfect treatment from which more than 15,000 new children will begin to suffer in the next year. The disease results from incorrect function of the patient´s immune system and thus therapies that target and correct disordered immunity must be sought. In this proposal, we will develop a roadmap for the innovative application of the regulatory power of B lymphocytes to overcome this disease
Keywords: Academic Medical Centers; Address; Adoptive Transfer; Allogenic; American; Antigen Presentation; Antigen-Presenting Cells; Antigens; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Autoimmunity; B cell repertoire; B-Cell Development; B-Lymphocyte Subsets; B-Lymphocytes; Beta Cell; career; career development; CD3 Antigens; Cell physiology; Cells; cellular targeting; Child; Childhood; clinically relevant; Data; Dedications; Development; Diabetes Mellitus; Disease; disorder risk; Environment; Evoked Potentials; Exhibits; Failure (biologic function); Fostering; Frequencies (time pattern); Functional disorder; Genetic; Human; Hyperglycemia; Immune; Immune system; Immune System Diseases; Immune Tolerance; Immunity; Immunobiology; Immunology; Inbred NOD Mice; Individual; Injury; innovation; Insulin; insulin dependent diabetes mellitus onset; Insulin-Dependent Diabetes Mellitus; Investigation; islet; Islet Cell; Knowledge; Life; Lymphocyte; Maintenance; Mediating; Mentors; Methods; Microbiology; Modeling; Molecular; Molecular Profiling; Monitor; Mouse Strains; Mus; novel; Organ; Outcome; Pathogenesis; Patients; pediatric department; Peripheral; peripheral tolerance; Physicians; planetary Atmosphere; Play; prevent; Process; Production; programs; public health relevance; Receptor Signaling; Regulation; Research Design; Residual state; Resistance; Resources; rituximab; Role; Scientist; Seminal; Signal Transduction; Skin; Specificity; Stimulus; System; T-Lymphocyte; Therapeutic; Toll-like receptors; Training; Transgenic Animals; Transplantation; Transplantation Tolerance; Universities
Project start date: 2011-05-01
Project end date: 2014-02-28
Budget start date: 1-MAY-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-10-059
1K08DK090146-01 (2011): $144600
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to J Daniel
J Daniel, Professor
Boston Universitycity: Boston country: United States (us)
Grant 5R01HG001389-16 from National Human Genome Research Institute
Abstract: We will develop methods and parallel flow instrumentation for high-throughput, high- content, cell-based screening, for phenotyping and manipulation of small rare-cell fractions, and for multiplexed expression analysis from 1-100 cells in flow. We will provide means for combinatorial scaling of mechanistic and small-molecule studies of protein misfolding disease, particularly neurodegenerative diseases such as Parkinson´s (model organism yeast). And for high-content cytometry studies with (rare) primary hematopoietic stem cells (mouse). This program would develop new instrumentation and methods for cell-based high- throughput, high-content screening (HCS). The program addresses bottlenecks that severely limit HCS for (i) scale-up to drug discovery, (ii) for handling of small samples of highly heterogeneous primary cell types; (iii) for finding rare cells (e.g. finding pluripotent cells for cancer diagnostics) and, (iv) image-based sorting/enrichment
Keywords: 3T3 Cells; Address; Algorithms; Amyloid; Animal Model; Architecture; base; Biological Assay; Cancer Diagnostics; Cell Fraction; Cell Separation; cell type; Cells; Collaborations; Color; combinatorial; Computer software; cost; Cytometry; Data; data reduction; design; Detection; detector; Disease; drug discovery; Flow Cytometry; Fluorescence; Funding; Goals; Hematopoietic stem cells; Hour; Image; Institutes; instrument; instrumentation; Lasers; Mammalian Cell; Measurement; Methods; Microfluidics; Microscope; models and simulation; Mus; Neurodegenerative Disorders; Nuclear Translocation; Parkinson Disease; Phenotype; Principal Investigator; Procedures; programs; protein misfolding; Proteins; public health relevance; RNA Interference; Sample Size; Sampling; scale up; Scanning; Schedule; Screening procedure; small molecule; Sorting - Cell Movement; Stem cells; Swiss 3T3 Cells; System; Texas; Time; United States National Institutes of Health; Universities; Validation; Work; Yeasts
Relevance: This program would develop new instrumentation and methods for cell-based high- throughput, high-content screening (HCS). The program addresses bottlenecks that severely limit HCS for (i) scale-up to drug discovery, (ii) for handling of small samples of highly heterogeneous primary cell types; (iii) for finding rare cells (e.g. finding pluripotent cells for cancer diagnostics) and, (iv) image-based sorting/enrichment
Project start date: 1995-09-30
Project end date: 2013-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-07-070
5R01HG001389-16 (2011): $709603
TRANSLATIONAL RESEARCH TRAINING IN SLEEP MEDICINE
J Daniel
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5T32HL082610-05 from National Heart, Lung, And Blood Institute
Abstract: This is a revised application of T32 HL082610, "Translational Research Training in Sleep Medicine." In response to the previous review, we have reorganized the Program directorship, added experienced biostatistician mentors, and addressed each of the reviewers´ other comments. The goal of this Program is to train the future generation of clinical and basic researchers in a translational approach to Sleep Medicine. Sleep disturbances produce wide-ranging morbidities in cardiopulmonary, metabolic, and neurological systems. Despite recent scientific breakthroughs in the basic science of sleep and circadian rhythms, there are few clinically-oriented investigators trained in the translational science of Sleep Medicine. Developing a successful training program in translational Sleep Medicine requires a foundation of vigorous multidisciplinary collaborations to represent the diversity of scientific perspectives in the field. Such collaborations exist at the University of Pittsburgh. A critical mass of collaborating researchers from three schools (Medicine, Public Health, Nursing) and six Departments or Divisions within the School of Medicine (Psychiatry, Pulmonary Medicine, Renal Medicine, Neurology, Behavioral Medicine, Neuroscience) currently receive external funding for sleep research. In addition, the University has an outstanding infrastructure and academic resources in sleep research including the Clinical Neuroscience Research Center, the Sleep and Circadian Rhythms Laboratory, two field centers within the Department of Epidemiology, and a basic research laboratory specializing in animal models of sleep disorders. The Training Program´s primary focus will be on post-doctoral training, with a secondary focus on mentored medical student research. The faculty and resources of the University will initially train two postdoctoral fellows per year, building to four trainees by the third year. The Program is intended primarily for M.D. scientists recruited from the fields of Pulmonary Medicine, Psychiatry, Neurology, and Internal Medicine. The core of the Training Program will be a mentored research experience that will be multidisciplinary in nature, translational in focus, grounded in a thorough understanding of sleep physiology, and structured by performance-based milestones. The Program will feature initial immersion in an eight week "basic training" course in animal and human sleep physiology, ongoing emphasis on presentation and publication of research findings, individually-prescribed coursework, and preparation of a career development award (K-series applications). The Program will include scheduled feedback and evaluation of both trainees and faculty mentors, and will be overseen by a Research Advisory Group consisting of Department Chairs and other T32 Program Directors, as well as an External Advisory Board. The medical student research experiences are designed to stimulate interest in Sleep Medicine specifically, and to address the critical "pipeline" issue facing translational research in a more general sense
Keywords: Medicine; Research Training; Sleep; Translational Research
Project start date: 2007-07-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-02-109
5T32HL082610-05 (2011): $199961
TESTING A NEUROBIOLOGICAL MODEL OF PRIMARY INSOMNIA
J Daniel
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5R01MH024652-36 from National Institute Of Mental Health
Abstract: This is a revised application of MH024652, originally reviewed in October, 2006. Primary Insomnia (PI) is a prevalent and chronic health problem associated with substantial morbidity, including increased risk for mood disorders. Despite its prevalence and consequences, little is known about insomnia pathophysiology. In this revised competing continuation, we will build on three novel findings from the current funding period 1) Using ecological momentary assessment (EMA), we have identified a distinctive diurnal pattern of mood and arousal symptoms in PI subjects that differentiates them from good sleeper controls (GSC); 2) We demonstrated that regional brain glucose metabolism shows prominent diurnal variation in healthy adults, most likely reflecting the influence of the circadian system on brainstem-hypothalamic arousal centers; 3) We showed that , compared to GSC, PI had significantly higher whole brain metabolism during wakefulness and non-rapid eye movement (NREM) sleep, and smaller sleep-related reductions in regional metabolism throughout brainstem, limbic, and frontal systems. The general aim of this application is to test a refined model of insomnia neurobiology by tying together our novel observations with recent neuroscience discoveries in sleep-wake regulation. Our model views insomnia as a disorder of sleep-wake state regulation, reflected across symptom, physiological, and functional anatomic levels. In the proposed study we will focus on the diurnal variation we have observed in insomnia symptoms, and on homeostatic sleep-wake regulation, because of its potential relevance to behavioral treatments for PI that involve sleep restriction. We will test our model using three strategies 1) Examining diurnal variation and sleep-wake state as naturalistic neurobiological probes, obtaining FDG PET studies during morning (a.m.) wakefulness, evening (p.m.) wakefulness, and NREM sleep. 2) Correlating a.m. wakefulness, p.m. wakefulness, and NREM sleep regional metabolism with concurrent symptom reports and physiological data. 3) Using one night of total sleep deprivation (TSD) as an experimental probe to examine homeostatic sleep-wake regulation. We will study 33 adults with PI and 33 age and sex-matched GSC. We will test three following Specific Aims Specific Aim 1. To characterize diurnal patterns of regional brain metabolism in PI and GSC. Specific Aim 2. To examine relationships between regional brain metabolism and concurrent insomnia symptoms (waking and sleep-related) and physiological data (EEG, HRV) in PI. Specific Aim 3. To examine homeostatic sleep regulation in PI and GSC by comparing changes in regional brain metabolism during NREM sleep before and after TSD in each group. The proposed study will use novel methods to advance our understanding of the neurobiology of PI, including aspects of neurobiology that are relevant to behavioral treatments. Insomnia is a common and impairing health condition, but we know very little about its causes, particularly in terms of brain function. This study will examine brain function in individuals with and without insomnia during wakefulness and during sleep; will examine how brain function is related to daytime and sleep symptoms; and will study the response of these individuals to one night of sleep deprivation. Results of this study could help us to better understand how the brain functions during sleep and wakefulness in people with insomnia, and how behavioral treatments (such as sleep restriction) might work
Keywords: Adult; Affect; Affective; Age; Anatomy; Arousal; Behavior Therapy; Brain; brain metabolism; Brain Stem; Cerebrum; Characteristics; Chronic; Circadian Rhythms; Clinical; Cognition; Cognitive; cognitive system; Data; Deoxyglucose; disturbance in affect; Dorsal; Electroencephalography; falls; Fatigue; Frequencies (time pattern); Functional disorder; Funding; Glucose; glucose metabolism; Health; Hypothalamic structure; Impaired health; Individual; interest; intervention effect; Metabolic; Metabolism; Methods; Modeling; Mood Disorders; Moods; Morbidity - disease rate; Neurobiology; Neurosciences; non rapid eye movement; novel; Pattern; Physiological; Positron-Emission Tomography; Prevalence; Primary Insomnia; Process; Publishing; Recovery; Regulation; Reporting; response; Risk; sex; Sleep; Sleep Deprivation; Sleep Disorders; sleep regulation; Sleeplessness; Structure; Symptoms; System; Testing; Thalamic structure; Time; Variant; Wakefulness; Work
Project start date: 1977-12-01
Project end date: 2012-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-047
5R01MH024652-36 (2011): $485041
INTERDISCIPLINARY TRAINING PROGRAM IN COGNITIVE SCIENCE
J Daniel, Professor
University Of Minnesota Twin Citiescity: Minneapolis country: United States (us)
Grant 5T32HD007151-33 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: This is a renewal application for five years of continued support for an interdisciplinary training program in cognitive science. The program integrates biological and behavioral approaches in three focused research domains of greatest strength at the University of Minnesota-perception and action, learning and memory, and emotion and affect. These topics are currently among the most active areas in cognitive science, and all have direct applications to human health. Future major advances in these areas are likely to come from researchers who have expertise bridging multiple approaches, thus motivating our plans to provide interdisciplinary training. Each year, the program will train 12 predoctoral students. Trainees, specializing in one of the three research domains, will receive advanced training in at least two of four major approaches-developmental, behavioral, neurobiological, and computational-and will have co-advisors representing at least two of these approaches. Features of the training program ensuring interdisciplinary breadth include multiple laboratory exposure, weekly multidisciplinary colloquia, journal clubs and seminars, travel to conferences, instruction in the responsible conduct of research and career skills, and an annual research symposium. Our trainees will have opportunities for translational research, and to interact with undergraduate students in under represented groups. Ultimately, our goal is to continue to provide a multi-lab, and multi-advisor experience that will allow our trainees to move fluidly among topics, instrumentation, and advanced methods, to address challenging problems in cognitive science and human mental health. The training program will be administered through the Center for Cognitive Sciences, an interdisciplinary, inter- departmental unit with an existing infrastructure of facilities and programs geared to graduate training. The 32 preceptors from nine departments all have strong training and research credentials, and all have active, major laboratories. The Center has the unique opportunity to develop a training program with faculty expertise capable of interweaving our three research domains and four cross-cutting approaches
Keywords: Cognitive Science; Training Programs
Project start date: 1978-07-01
Project end date: 2014-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-06-468
5T32HD007151-33 (2011): $387270
DE-CELLULARIZED HUMAN LUNGS FOR EX VIVO LUNG REGENERATION
J Daniel, Associate Professor
University Of Vermont & St Agric Collegecity: Burlington country: United States (us)
Grant 1R21HL108689-01 from National Heart, Lung, And Blood Institute
Abstract: There will never be enough donor lungs available to meet current and future transplantation needs. In contrast, de-cellularization of whole cadaveric lungs will result in an intact scaffold that can be re- cellularized with adult stem cells, including induced pluripotent stem cells (iPS), derived from individual patients and subsequently utilized for autologous transplantation. Notably, the de-cellularization process removes cellular antigens responsible for immune rejection and the de-cellularized lungs maintain native airway and alveolar architecture, extracellular matrix protein composition, and pulmonary vascular network. This will provide a potentially limitless supply of unrelated donor cadaveric lungs for use in emphysema and other diseases for which there is currently no effective cure. However, there are several fundamental questions to be addressed for use of human lungs as scaffolds. Key among these is the reliability and reproducibility of the de-cellularization and re-cellularization processes. Unlike homogenous laboratory mice, donor human lungs will come from patients of different ages and from a variety of clinical and disease backgrounds, including a history of smoking, which might influence the nature of the scaffold following de-cellularization or the re-cellularization process itself. Therefore this proposal will focus on determining key differences between de-cellularized lung scaffolds originating from different donors and on discovering key environmental conditions that determine successful re-cellularization of lung scaffolds with human mesenchymal stromal cells (hMSCs) and induced pluripotent stem cells (iPS). Our preliminary data demonstrates that human lung can be successfully de-cellularized and studied by a variety of histological and functional assessments including lung mechanics, surfactant production, vascular perfusion, and epithelial barrier function, as the lungs re-cellularize. Further, preliminary data demonstrates that human mesenchymal stromal cells (hMSCs) and human alveolar epithelial cells (A549 cells) can be successfully inoculated and grown in de-cellularized human lung. These data provide a firm platform for the proposed two Specific Aims. 1) To determine key differences between de-cellularized lung scaffolds obtained from different donors. 2) To optimize conditions for growth and differentiation of hMSCs, and human iPS cells into functional three dimensional lung tissue when inoculated into de-cellularized human lung slices and apply this technology to whole de-cellularized human lungs. (End of )
Keywords: 3-Dimensional; A549; ing; Address; Adult; adult stem cell; Age; Alveolar; Antigens; Architecture; Autologous; Autologous Transplantation; Autopsy; Biomedical Engineering; Blood Vessels; Bone Marrow; cell type; Cells; Clinical; Culture Media; Data; Development; Differentiation and Growth; Disease; EGF gene; Electron Microscopy; embryonic stem cell; Endothelial Cells; Endothelium; Environmental Risk Factor; Epithelial; Epithelial Cells; Epithelium; Extracellular Matrix Proteins; Fibroblast Growth Factor; Fibroblasts; Future; Growth; Growth Factor; Hepatocyte; Histologic; Human; Hydrocortisone; Immune; Implant; Individual; induced pluripotent stem cell; keratinocyte; Laboratory mice; light microscopy; Lung; lung lobe; Manuscripts; Measurement; Measures; Mechanical ventilation; Mechanics; meetings; Mesenchymal; Mesenchymal Stem Cells; Methods; Mus; Natural regeneration; Nature; Oxygen measurement, partial pressure, arterial; Patients; Perfusion; Pneumonectomy; Process; Production; Proteomics; Pulmonary Emphysema; Pulmonary Fibrosis; Reproducibility; response; scaffold; Screening procedure; Services; Slice; Smoking History; Stromal Cells; Structure of parenchyma of lung; surfactant; System; Techniques; Technology; Testing; Tissues; Transplantation; Tretinoin; Vascular Endothelial Growth Factors; Vascular Endothelium
Relevance: There will never be enough donor lungs available to meet current and future transplantation needs. In contrast, de-cellularization of whole cadaveric human lungs will result in an intact scaffold that can be re-cellularized with adult stem cells, including induced pluripotent stem cells (iPS) derived from individual patients and subsequently utilized for autologous transplantation. Our preliminary data demonstrate that human lung can be successfully de-cellularized and re-cellularized with both adult stem cells and with lung epithelial cells. The proposed multi-institutional collaborative studies will utilize state-of-the-art bioengineering techniques to develop optimal methods for growing functional lung tissue in de-cellularized cadaveric human lungs. This will provide a potentially limitless supply of unrelated donor cadaveric lungs for use in diseases such as emphysema, pulmonary fibrosis, and others for which there is currently no effective cure
Project start date: 2011-08-18
Project end date: 2013-07-31
Budget start date: 18-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: RFA-HL-11-026
1R21HL108689-01 (2011): $182597
2/3-IMPROVING DEPRESSION OUTCOME BY ADDING CBT FOR INSOMNIA TO ANTIDEPRESSANTS
J Daniel
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5R01MH078961-04 from National Institute Of Mental Health
Abstract: The purpose of the proposed study is to improve depression outcome for individuals with major depressive disorder and co-morbid insomnia by combining state-of-the-art antidepressant algorithm (MED) and empirically supported cognitive-behavioral therapy for insomnia (CBTI). Insomnia is an important aspect of depression and is only incompletely addressed by existing treatments. As persistent insomnia is both a risk factor for depressive relapse and a more general indicator of disease severity and poor prognosis, identification of more effective therapies for depressed patients with insomnia has public health significance. Therefore, we propose a prospective, randomized controlled study that will test the efficacy of the proposed strategy in individuals with major depressive disorder and comorbid insomnia. The primary aim is to test whether the proposed intervention increases rates of remission from depression following 16 weeks of treatment compared with a control intervention. A secondary aim is to test whether the proposed intervention enhances longer-term outcome by reducing the proportion of participants who relapse during a 6-months follow-up phase, relative to the control treatment. Participants will be 300 men and women, drawn from 3 study sites, so that the sample is diverse with respect to race and ethnicity. Participants will meet DSM-IV criteria for Major Depressive Disorder, score 18 or higher on the 17-item Hamilton Depression Rating Scale (HRSD) and have difficulty initiating or maintaining sleep that is clinically significant (DSM-IV criteria for insomnia). After the end of the acute treatment phase, participants will be transitioned to community care and will be followed-up for 6 months. The primary outcome measures are the HRSD and the depression portion of the SCID-IV, to be administered by masked raters at baseline, biweekly during the acute phase, and monthly during the follow-up phase. Secondary measures include measures of sleep and functional outcome. Relevance Achieving and maintaining remission is the desired clinical goal for MDD. By evaluating the efficacy of a treatment strategy that combines a standardized antidepressant pharmacotherapy with a non- pharmacological therapy that targets insomnia (CBTI), the proposed study, could lead to clinically meaningful improvement in the lives of many patients with MDD and insomnia. Insomnia is a common problem in patients with major depression, and is associated with poor response to depression treatment, recurrence of depression, and increased risk for suicide. This study will examine whether a specific psychological-behavioral treatment for insomnia, in conjunction with state-of-the-art antidepressant medication treatment, improves the outcomes of patients with major depression and insomnia. The results of this study may offer a method for improving the treatment of major depression without the use of additional medication
Keywords: Acute; Address; Affect; Algorithms; Antidepressive Agents; Applications Grants; Attention; Behavior Therapy; Behavioral; Caring; Chronic; Clinical; clinically significant; Cognitive; Cognitive Therapy; Communities; Comorbid Insomnia; Control Groups; Depressed mood; depressive symptoms; Diagnostic; disability; Disease; Disease remission; DSM-IV; effective therapy; efficacy testing; Escitalopram; Ethnicity aspects; experience; follow-up; functional outcomes; Goals; Hamilton Rating Scale for Depression; Healthcare Systems; improved; Individual; Intervention; Lead; Major Depressive Disorder; Masks; Measurement; Measures; meetings; men; Mental Depression; Methods; novel strategies; Outcome; outcome forecast; Outcome Measure; Oxalates; Participant; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase; Pilot Projects; Placebos; Population; primary outcome; prospective; psychologic; Psychological desensitization therapy; Psychotherapy; public health medicine (field); Quality of life; Race; Randomized Controlled Trials; Recurrence; Relapse; Relative (related person); Reporting; response; Risk Factors; Sampling; Severities; Severity of illness; Site; Sleep; Sleeplessness; suicidal risk; Symptoms; Testing; Time; treatment duration; Treatment outcome; treatment strategy; Woman
Project start date: 2008-06-01
Project end date: 2013-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-092
5R01MH078961-04 (2011): $231847
FUNCTIONAL ANALYSIS OF THE PROTEASOME BASE
J Daniel
Harvard University (medical School)city: Boston country: United States (us)
Grant 5R37GM043601-21 from National Institute Of General Medical Sciences
Abstract: The proteasome, the most complex proteolytic assembly known, is a key mediator of biological regulation in eukaryotes. Ubiquitinated substrates are recognized by the 900-Kda regulatory particle (RP), then unfolded and translocated through a gated channel into the interior chamber of the core particle (CP) to be degraded. The key steps of recognition and unfolding are poorly understood, but appear to be critically dependent on a subassembly of the RP known as the base. The base is in direct contact with the CP, and is composed of 8 subunits, 6 of them ATPases of the AAA family. The present proposal aims to better understand how the base promotes protein degradation. Substrate recognition appears to be mediated by an ensemble of ubiquitin chain binding factors, including Rpnl0 and the UU proteins (such as Rad23 and Dsk2), all of which associate with the base. Still other factors, possibly intrinsic to the base, are also likely to be involved. In Aim 1 we will take a combined genetic, biochemical, and proteomic approach to resolve how these multiple pathways of substrate recognition are related do they function redundantly, cooperatively, or independently? Are their functions overlapping for some substrates while being uniquely required to recognize others? The main focus of Aim 2 is an almost completely unstudied part of the base, the N-terminal domains of the 6 ATPases. Our hypothesis is that the N-domains play key roles in protein degradation, by projecting into the central compartment of the RP, where unfolding is thought to occur, and helping to promote this event by interacting directly with substrate. Among our proposed tests of this idea are attempts to screen genetically for mutations in the N-domains that differentially impair the degradation of one substrate but not another, at a step (presumably unfolding) that follows ubiquitin chain recognition. Aim 3 addresses our hypothesis that the largest subunit of the base, Rpnl, acts as a scaffold to recruit multiple factors to the proteasome, all of which are active on multiubiquitin chains. The proposed multiple functions of Rpnl will be dissected mainly by obtaining point mutants that specifically abrogate individual binding sites for postulated ligands such as Rad23, Rpnl0, and Ubp6. In summary, this proposal will combine biochemical, genetic, proteomic, and structural methods to address key questions concerning early steps in protein degradation by the proteasome
Keywords: Address; Affect; ATP phosphohydrolase; base; Binding (Molecular Function); Binding Sites; Biochemical; Biochemical Genetics; Biological; Complex; Data; Deletion Mutation; Electron Microscopy; Eukaryota; Event; Family; Genetic; In Vitro; Individual; Ligand Binding; Ligands; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Methods; multicatalytic endopeptidase complex; mutant; Mutation; N Domain; N-terminal; Nucleosome Core Particle; particle; Pathway interactions; Pattern; Phenocopy; Play; Process; Property; protein degradation; Proteins; Proteomics; Recruitment Activity; Regulation; scaffold; Scaffolding Protein; Testing; Ubiquitin; Ubiquitination
Project start date: 1989-12-01
Project end date: 2014-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5R37GM043601-21 (2011): $568309
5R37GM043601-20 (2010): $613159
STUDIES OF USP14 AND THE UBIQUITIN STRESS RESPONSE
J Daniel
Harvard University (medical School)city: Boston country: United States (us)
Grant 5R21DK082906-02 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: The proteasome is a key enzyme in the ubiquitin-dependent pathway of degradation. It is important in human health and is responsible for proper regulation of a wide variety of basic cell functions. Historically, little attention has been given to how the proteasome is regulated. Our recent studies in yeast have identified unanticipated mechanisms of proteasome regulation. The two mechanisms involve a proteasome-associated deubiquitinating enzyme, Ubp6, whose mammalian ortholog is Usp14 (i) Ubp6 inhibits the activity of the proteasome, and (ii) the expression of Ubp6, and in turn the amount of Ubp6 on the proteasome, are negatively regulated by ubiquitin levels. These two mechanisms are likely related, in that low ubiquitin levels induce Ubp6, which then inhibits proteasome activity, which has the effect of reducing the destruction of scarce ubiquitin. Our preliminary data indicate that, as predicted from our findings in yeast, Usp14 inhibits mammalian proteasomes. In Aim 1, we will characterize the mechanism of this inhibition in mammals and its effects on cell function, using purified proteasomes and purified Usp14 as well as an existing line of usp14 null MEF cells. In Aim 2, we will attempt to isolate the molecular machinery of the ubiquitin stress response. We present preliminary data that the ubiquitin stress response, which we first identified in yeast, may operate in mammals as well. We will use both yeast and mammalian systems to dissect the response. A large number of diseases involve protein misfolding and its pathological effects. It is well appreciated that misfolded proteins are preferred substrates for the ubiquitin-proteasome pathway. Thus, in many disease states, misfolded proteins may impose an unusually high load on the ubiquitin-proteasome system. In such instances, it may be necessary for the system to maintain function through compensatory regulatory mechanisms. Therefore, the types of regulatory mechanisms that will be probed here for the first time in mammals may potentially be relevant to the pathogenesis of multiple diseases. PUBLIC HEALTH RELEVANCE The proteasome is a key enzyme that controls the levels of hundreds of proteins and thus is responsible for the proper regulation of a wide variety of basic cell functions. Our recent studies in yeast have identified unanticipated mechanisms by which this enzyme is itself controlled. The grant is designed mainly to test whether these interesting new mechanisms may apply to mammals. If so, the work may advance our understanding of human health, because the ubiquitin-proteasome system plays important roles in both cancer and neurodegenerative disease
Keywords: 20S Catalytic Proteasome; 20S Core Proteasome; 20S Proteasome; 20S Proteosome; aberrant protein folding; abnormal protein folding; APF-1; Assay; Ataxia; Ataxy; ATP-Dependent Proteolysis Factor 1; Attention; Bioassay; Biologic Assays; biological adaptation to stress; Biological Assay; Cancers; Cell Function; Cell physiology; Cell Process; Cells; Cellular Function; Cellular Physiology; Cellular Process; cis acting element; Coordination Impairment; Data; Degenerative Diseases, Nervous System; Degenerative Neurologic Disorders; Degradation Pathway; Degradative Pathway; design; designing; Deubiquitinating Enzyme; Deubiquitination; Disease; disease/disorder; Disorder; Dyssynergia; Elements; Enzymes; experiment; experimental research; experimental study; gene product; Grant; Health; High Mobility Protein 20; HMG-20; Human; Human, General; Hybrids; in vitro testing; in vivo; interest; Laboratories; Macropain; Macroxyproteinase; malignancy; Malignant Neoplasms; Malignant Tumor; Mammalia; Mammalian Cell; Mammals; Mammals, General; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Mediating; Messenger RNA; Mice; Mice, Mutant Strains; Modeling; Molecular; mouse mutant; mRNA; multicatalytic endopeptidase complex; Multicatalytic Proteinase; Murine; Mus; mutant; Mutant Strains Mice; Nature; neoplasm/cancer; Neurodegenerative Diseases; Neurodegenerative Disorders; neurodegenerative illness; Neurologic Degenerative Conditions; Neurologic Diseases, Degenerative; novel; Ortholog; Orthologous Gene; Pathogenesis; pathologic protein folding; pathway; Pathway interactions; Phenotype; Play; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Prosome; Proteasome; Proteasome Endopeptidase Complex; Proteasome Inhibition; protein degradation; Protein Degradation, Metabolic; Protein Degradation, Regulatory; protein mis-folding; protein misfolding; Protein Turnover; Proteins; Proteosome; public health relevance; reaction; crisis; reconstitute; reconstitution; Regulation; Regulatory Element; RegulatoryElement; research study; response; RNA, Messenger; Role; screening; Screening procedure; screenings; social role; stress response; stress; reaction; Subcellular Process; System; System, LOINC Axis 4; Testing; Time; transcription factor; Ubiquitin; Work; Yeasts
Relevance: The proteasome is a key enzyme that controls the levels of hundreds of proteins and thus is responsible for the proper regulation of a wide variety of basic cell functions. Our recent studies in yeast have identified unanticipated mechanisms by which this enzyme is itself controlled. The grant is designed mainly to test whether these interesting new mechanisms may apply to mammals. If so, the work may advance our understanding of human health, because the ubiquitin-proteasome system plays important roles in both cancer and neurodegenerative disease
Project start date: 2008-09-30
Project end date: 2011-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2011
PFA/PA: PA-06-181
5R21DK082906-02 (2009): $211875
PREGNANCY AND RESPONSE TO ANTIRETROVIRAL THERAPY IN SOUTH AFRICA
J Daniel, Assistant Professor
Duke Universitycity: Durham country: United States (us)
Abstract: The overall goal of this proposal remains unchanged and is to investigate the effect of pregnancy on response to highly active antiretroviral therapy (HAART) among HIV-i- women in treatment in Johannesburg, South Africa. In addition, we will continue developing the advanced quantitative epidemiologic methods described in the training component ofthe K99 portion of this grant. Prevalence of HIV among pregnant women in South Africa is substantially higher than that among other adults. Despite extensive study of interventions for the prevention of mother to child transmission of HIV, and response to antiretroviral therapy during and after such interventions, little is known about the effect of pregnancy itself on responses to and retention in therapy. Fully understanding the effect of pregnancy on maternal responses to HAART is essential to providing good clinical care to H1V+ women in Africa and elsewhere. To address these questions, this work proposes to examine the effect of pregnancy on immunologic responses to HAART, clinical (AIDS/death) responses to HAART, and to adherence to HAART and retention in care. Because it is impossible (and highly unethical) to randomize the exposure of pregnancy, this study will rely on high quality clinical data collected in the Themba Lethu Clinic in Johannesburg, South Africa, an urban antiretroviral therapy clinic associated with the Clinical HIV Research Unit at the University ofthe Witwatersrand. The Principal Investigator of the proposed work, Dr. Daniel Westreich, has a strong track record in producing high quality research in collaboration with investigators and clinicians at the Clinic. The analysis of these data will require the use of structural models, an advanced body of epidemiologic methods
Keywords: Acquired Immunodeficiency Syndrome; Address; Adherence (attribute); Adult; Africa; Africa South of the Sahara; Age; antiretroviral therapy; attenuation; base; Caring; CD4 Lymphocyte Count; Cessation of life; Clinic; Clinical; clinical care; Clinical Data; Clinical Treatment; Collaborations; Counseling; Data; Data Analyses; Epidemiologic Methods; Epidemiology; evidence base; experience; Female of child bearing age; follow-up; Goals; Grant; Health; Health Policy; Highly Active Antiretroviral Therapy; HIV; HIV Infections; Immunologics; Instruction; Intervention; Intervention Studies; Investigation; Life; men; Mentors; Methods; Phase; Pregnancy; pregnant; Pregnant Women; Prevalence; Preventive Intervention; Principal Investigator; protective effect; public health medicine (field); Randomized; reproductive; Research; Research Personnel; response; Risk; Scientist; South Africa; Structural Models; Time; Training; Universities; Vertical Disease Transmission; Woman; Work
Relevance: RELEVANCE (See instructions): This work will answer a suite of key substantive questions around the impact of pregnancy on response to HAART in South Africa. In doing so, this work will dramatically increase the base of scientific and public health evidence on this important subject, most likely in a way which is generalizable to much of sub-Saharan Africa. In addition, this work and training will continue to develop and establish Dr. Westreich as a health scientist
Project start date: 2010-09-27
Project end date: 2013-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
5R00HD063961-03 (2011): $0
Sponsored Links Excellgen http://Excellgen.com
STATISTICAL LEARNING OF MULTIPLE PATTERNS IN INFANTS, ADULTS, AND MONKEYS
J Daniel, Assistant Professor
Pennsylvania State University-univ Parkcity: University Park country: United States (us)
Grant 1R01HD067250-01A1 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: The overall goal of the present grant application is to understand how a naive learner collects distributional information from the environment and makes an implicit decision that the corpus of input contains either a single structure or multiple structures. Mature learners are incredibly facile at interpreting information in a context-specific manner, thereby partitioning the input into two or more sub-structures. We will investigate this question of context-specific statistical learning by studying two types of naove learners - human infants and tamarin monkeys - as well as mature adults. The specific objective of the proposed research is to determine whether and how infants learn that there are multiple patterns of information embedded in streams of speech, or that there are multiple words that refer to the same object, and to determine whether context-specific statistical learning has species-specific biases. Two types of experimental designs will be used to study context-specific statistical learning. The first uses a single change in the underlying structure. A variety of contextual cues will be introduced to signal that the underlying structure has undergone a change, and the dependent measure is whether the learner has acquired the first, the second, both the first and the second, or neither structures. The second design uses two alternating structures that are signaled by a variety of stimulus cues to partition the two underlying structures. It is important to note that in both of these designs, if the learner aggregates the structural information across the entire corpus, rather than partitioning the corpus into two subsets, no learning is possible. Thus, these designs test the ability of the learner to extract the contextual cues that partition the input into subsets. The implications of the proposed studies are fundamental to any theory of learning, but particularly to the kind of implicit (passive exposure) statistical learning that is thought to characterize much of early human development in many domains. Infants must learn - by a combination of sensitivity to distributional patterns and innate biases - that patterns of information are context-specific, as in the case of bilingualism. Our proposed experiments will extend our recent studies of human adults by determining (a) whether infants show the same pattern of learning biases (primacy effects) and context-sensitivity (to talker voice), (b) whether tamarin monkeys show these same biases and context effects, and (c) what the limits of context-specific statistical learning are in human adults and infants in both word segmentation and referential tasks. Language development is one of the hallmarks of the human species, yet it is difficult to study because of the huge variation in early exposure to different amounts of linguistic input. The use of artificial languages that are acquired in the lab over a few hours provides a window on the mechanisms of language development. We will study language learning in the lab to gain a unique perspective on how the infants and adults learn the patterns of words in streams of speech and contrast this with performance in nonhuman primates. These studies will not only reveal a basic mechanism of language learning, but also establish benchmarks against which language delay can be compared. Moreover, understanding the mechanisms that lead to successful acquisition in normal infants and adults can help to identify loci of language disorders and design methods for remediating disorders
Keywords: Adult; American; Applications Grants; auditory stimulus; Benchmarking; bilingualism; British; Cues; Data; design; Development; Disease; Economics; Elements; Environment; Experimental Designs; Exposure to; Eye; Goals; Head; Head Movements; Hour; Human; Human Development; Infant; Judgment; Language; Language Delays; Language Development; Language Disorders; Lead; Learning; lexical; Linguistics; Location; Machine Learning; man; Measures; Methods; Monkeys; nonhuman primate; novel; Parents; Pattern; Performance; Phonetics; phonology; preference; Procedures; public health relevance; Recovery; Research; research study; response; Rivers; Role; Saguinus; Semantics; Signal Transduction; Speech; Stimulus; Stream; Structure; Sum; System; Testing; theories; Time; Variant; Voice; Work
Relevance: Public Health Relevance Statement Language development is one of the hallmarks of the human species, yet it is difficult to study because of the huge variation in early exposure to different amounts of linguistic input. The use of artificial languages that are acquired in the lab over a few hours provides a window on the mechanisms of language development. We will study language learning in the lab to gain a unique perspective on how the infants and adults learn the patterns of words in streams of speech and contrast this with performance in nonhuman primates. These studies will not only reveal a basic mechanism of language learning, but also establish benchmarks against which language delay can be compared. Moreover, understanding the mechanisms that lead to successful acquisition in normal infants and adults can help to identify loci of language disorders and design methods for remediating disorders
Project start date: 2011-04-01
Project end date: 2016-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-10-067
1R01HD067250-01A1 (2011): $221561
GENOMIC REGULATION OF CHROMATIN ACCESSIBILITY DURING DROSOPHILA DEVELOPMENT
J Daniel
University Of North Carolina Chapel Hillcity: Chapel Hill country: United States (us)
Grant 5F32GM090759-02 from National Institute Of General Medical Sciences
Abstract: The organization of the genome into chromatin plays a central role in the regulation of developmental gene expression programs. One widely held belief to explain this regulation is the coordinated control of chromatin accessibility. However, there is little high resolution data measuring chromatin accessibility in vivo, the regulators of large scale chromatin accessibility are largely unknown, and the molecular mechanisms by which chromatin accessibility affects gene activity are unclear. This proposal describes a genomics approach to directly measure chromatin accessibility at high resolution in order to develop a system in the experimentally tractable Drosophila model that will examine the relationship between chromatin accessibility and gene activity. To accomplish this objective, we will utilize a technique developed by the Lieb lab termed FAIRE (Formaldehyde-Assisted Isolation of Regulatory Elements). We present data here not only demonstrating the ability of FAIRE to reproducibly isolate discrete regions of ´open´ chromatin, but also to identify broad chromosomal regions that share chromatin accessibility properties, even using small numbers of cells. Specifically, we will first perform a genomics screen to identify novel regulators of chromatin accessibility by using RNAi-mediated knockdown in Drosophila S2 cells, followed by analysis of FAIRE-enriched DNA on microarrays. Second, we will generate genomic chromatin accessibility maps for different tissues and stages of Drosophila development by performing FAIRE followed by high-throughput sequencing, providing a framework to assess the relationship between chromatin accessibility and gene activity during development and identifying cis-regulatory modules that control developmental expression programs. Finally, we will begin to characterize potential regulators of chromatin accessibility in vivo through mutational analyses. Relevance to Public Health Many developmental pathways and mechanisms, when misregulated, contribute to human disease. Therefore, the knowledge gained by these efforts will have applications in understanding mechanisms that underlie disease states such as cancer
Keywords: Adult; Affect; Belief; Biology; Cell Count; cell determination; Cell division; Cell Nucleus; cell type; Cells; Chromatin; Chromatin Structure; Collection; Communities; Data; Development; Developmental Gene Expression Regulation; Digestion; Disease; DNA; Double-Stranded RNA; Drosophila genus; Drosophila inturned protein; Embryo; Euchromatin; follow-up; Formaldehyde; Gene Expression Regulation; Genes; Genome; genome-wide; Genomics; Heterochromatin; human disease; in vivo; knock-down; Knowledge; Malignant Neoplasms; Maps; Measures; Mediating; Microscopy; Modeling; Molecular; Molecular Genetics; novel; nuclease; Organism; Pathway interactions; Play; Polycomb; Process; programs; Property; Proteins; public health medicine (field); Regulation; Regulatory Element; Research; Resistance; Resolution; Resources; RNA Interference; Role; Specific qualifier value; Staging; System; Techniques; Time; Tissues; Validation; Work
Project start date: 2010-04-01
Project end date: 2011-09-30
Budget start date: 1-APR-2011
Budget end date: 30-SEP-2011
5F32GM090759-02 (2011): $25237
HOMING AND HOMEOSTASIS OF REGULATORY T CELLS
J Daniel
Benaroya Research Inst At Virginia Masoncity: Seattle country: United States (us)
Grant 5R01AI067750-05 from National Institute Of Allergy And Infectious Diseases
Abstract: The objective of this study is to determine how specific combinations of homing receptors control the localization and function of CD4+CD25+ regulatory T cells (Treg). As potent modulators of self-reactive T cells, Treg represent an exciting new therapeutic approach for the treatment of autoimmune disorders such as type 1 diabetes, multiple sclerosis, rheumatoid arthritis and Chron´s disease. However, as a prerequisite to understanding how Treg function in vivo to control autoimmunity, it is essential to determine where Treg localize and what cell types they interact with. While Treg express diverse and heterogeneous patterns of lymphocyte homing receptors, the relationship between their homing receptor expression, their localization, and their ability to modulate organ-specific autoimmunity has not been explored experimentally. We hypothesize that Treg must localize to and function within specific lymphoid and non-lymphoid tissues in order to prevent autoimmunity; Here we propose a series of experiments to test this hypothesis, and determine how disrupting Treg localization impacts their functional ability to prevent systemic and organ-specific autoimmunity (aim 1) and their survival/homeostasis (aim 2). In addition, we will test the hypothesis that homing receptor expression defines Treg subsets targeted to lymphoid vs. non-lymphoid tissues that have distinct functional and homeostatic characteristics (aim 3). Determining the relationship between Treg homing and their ability to control autoimmunity is required to understand where these cells function in vivo, and how diverse populations of Treg may function at distinct sites to prevent the initiation and/or progression of autoimmunity. This has clear and direct implications in the clinical application of Treg to the prevention and treatment of human autoimmune and inflammatory diseases
Keywords: Activities of Daily Living; allograft rejection; Autoimmune Diseases; Autoimmune Process; Autoimmunity; base; CD4 Positive T Lymphocytes; Cell physiology; cell type; Characteristics; clinical application; Disease; Environment; Equilibrium; Gene Targeting; Goals; Homeostasis; Homing; Human; immunopathology; in vivo; Inflammatory; innovation; Insulin-Dependent Diabetes Mellitus; Lymphocyte Homing Receptors; Lymphoid; Lymphoid Tissue; migration; Molecular; Multiple Sclerosis; Mus; novel; novel therapeutic intervention; Organ; pathogen; Pattern; Play; Population Heterogeneity; prevent; Prevention; Property; receptor; receptor expression; Regulatory T-Lymphocyte; Research; research study; response; Rheumatoid Arthritis; Role; Secondary to; Series; Site; System; T-Lymphocyte; Testing; Tissues; trafficking
Project start date: 2006-05-15
Project end date: 2011-10-30
Budget start date: 1-MAY-2010
Budget end date: 30-OCT-2011
5R01AI067750-05 (2010): $431435
ADIPONECTIN REGULATION OF THE MESOLIMBIC DOPAMINE SYSTEM
J Daniel
University Of Texas Hlth Sci Ctr San Antcity: San Antonio country: United States (us)
Grant 1R21MH096251-01 from National Institute Of Mental Health
Abstract: The mesolimbic dopamine system is a major neural substrate for reward, motivation and emotion. Dysfunction of this system has been linked to several psychiatric conditions including eating disorders, drug abuse, schizophrenia and depression. Recent studies suggest adiposity status and adipose-derived hormones, termed "adipokines", modulate motivational and emotional responses. Adiponectin is the most abundant adipokine in circulation that can access the central nervous system. Our preliminary studies demonstrate that adiponectin induces activation of dopamine neurons in the ventral tegmental area (VTA) and that adiponectin receptor 1 (AdipoR1) is expressed in VTA dopamine neurons, suggesting that adiponectin functionally interacts with the mesolimbic dopamine system. The proposed studies in this project are to determine whether adiponectin regulates mesolimbic dopamine activity via interacting with AdipoR1 on dopamine neurons. To test this, we will first determine the effect of exogenous adiponectin on firing properties of VTA dopamine neurons and dopamine release in the nucleus accumbens, one of the major targets of the VTA dopamine neurons. Second, we will generate conditional knockout mice lacking AdipoR1 specifically in dopamine neurons and determine firing properties of VTA dopamine neurons as well as dopamine efflux in these mutant mice. These studies will provide novel information on the regulation of the mesolimbic dopamine pathway and new insights into dopamine-related processes in psychiatric illnesses especially those associated with metabolic syndrome such as obesity and diabetes. Abnormal dopamine-related processes lead to aberrant reward, motivation and emotion in psychiatric disorders including eating disorders, drug addition, schizophrenia and depression. The proposed studies will enhance our understanding of how the mesolimbic dopamine system is regulated by the adipose-derived hormone adiponectin and will have clinical implications for psychiatric disorders associated with obesity and type 2 diabetes
Keywords: Address; adipokines; adiponectin; Adipose tissue; Area; base; Behavior; Biological; biomarker; Blood Circulation; Clinical; Diabetes Mellitus; Disease; Dopamine; dopamine system; dopamine transporter; dopaminergic neuron; Drug abuse; Eating Disorders; Emotional; Emotions; Endocrine Glands; FOS gene; Functional disorder; Hormones; in vivo; Injection of therapeutic agent; insight; Knockout Mice; Lead; Leptin; Link; Measures; Mediating; Mental Depression; Mental disorders; mesolimbic system; Metabolic syndrome; Microdialysis; Microinjections; Motivation; Mus; Mutant Strains Mice; Neuraxis; neurochemistry; Neurons; neuropsychiatry; Non-Insulin-Dependent Diabetes Mellitus; novel; Nucleus Accumbens; Obesity; Pathway interactions; Pattern; peptide hormone; Peptides; Pharmaceutical Preparations; Population; Process; Property; receptor; Regulation; relating to nervous system; response; Rewards; Role; Schizophrenia; Signal Transduction; System; Testing; Transgenic Mice; Ventral Tegmental Area
Relevance: Abnormal dopamine-related processes lead to aberrant reward, motivation and emotion in psychiatric disorders including eating disorders, drug addition, schizophrenia and depression. The proposed studies will enhance our understanding of how the mesolimbic dopamine system is regulated by the adipose-derived hormone adiponectin and will have clinical implications for psychiatric disorders associated with obesity and type 2 diabetes
Project start date: 2012-02-22
Project end date: 2014-01-31
Budget start date: 22-FEB-2012
Budget end date: 31-JAN-2013
1R21MH096251-01 (2012): $221330
MARC: MINORITY ACCESS TO RESEARCH CAREERS
J Daniel, Lecturer
State University New York Stony Brookcity: Stony Brook country: United States (us)
Grant 5T34GM008655-13 from National Institute Of General Medical Sciences
Abstract: The long-term goal of this project is to increase the numbers of underrepresented minority students who obtain their doctorate and pursue research careers in the biomedical sciences. The basic strategy is to continue to develop and maintain partnerships with schools that have large numbers of underrepresented students, to recruit and select highly qualified students, to provide them with quality laboratory experiences and challenging courses during their undergraduate years, to provide them with financial and academic support services that maximize their potential for success, and to provide them with the qualifications and references so they may compete for entry into the strongest graduate programs in the country. The objectives of the MARC program are 1) to attract underrepresented students to biomedical disciplines and courses of study at Stony Brook; 2) to identify underrepresented students at Stony Brook with an interest and potential in pursuing research careers in the biomedical sciences and related fields early in their academic tenure; 3) to motivate underrepresented students to obtain degrees in biomedical sciences and related fields; 4) to help students gain confidence in their abilities by early exposure to laboratory experiences, research, public speaking and biomedical researchers; 5) to create a positive environment through the use of group activities, mentoring and advising; 6) to promote an awareness of the opportunities available in biomedical research through peer, graduate student and faculty mentoring, as well as research experiences at other institutions; and 7) to create a national model for outreach and undergraduate education at Stony Brook that will ensure a diverse and highly-trained biomedical research workforce for the future
Keywords: Awareness; Biomedical Research; career; CCL7 gene; Country; Discipline; Educational aspects; Ensure; Environment; experience; Exposure to; Faculty; Future; Goals; graduate student; Institution; interest; Laboratories; Mentors; Minority; Modeling; outreach; peer; programs; Public Speaking; Qualifying; Recruitment Activity; Research; Research Personnel; Schools; Science; Services; Students; success; Training; Underrepresented Minority
Project start date: 1997-06-01
Project end date: 2014-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PAR-07-337
5T34GM008655-13 (2011): $205631
J Daniel, Prof Of Pathology & Laboratory Medicine
Emory Universitycity: Atlanta country: United States (us)
Abstract: The Human Tissue and Pathology Core (Shared Resource Category 4.06) of the Winship Cancer Institute (WCI) will be critical to the overall clinical and scientific success by enhancing tissue-based investigation through its procurement and distribution of human cancer specimens and its comprehensive histopathologic services. The Core is highly integrated within the WCI, serving individual and team science projects in all of the major programs and interfacing directly with the other Cores. The tissue banking component of the Core consists of an IRB-approved Human Tissue Procurement Service (HTPS), located within the Emory University Hospital and the WCI, with affiliated banking sites at Grady Memorial and Crawford Long Hospitals. Tissue procurement personnel are highly experienced in anatomic pathology and tumor banking protocols, with quality assurance of specimens provided by members of the Department of Pathology. Tumor and normal control tissues, along with other biospecimens associated with specialized protocols, are procured as frozen, fresh, fixed, embedded or other specialized preparation, and inventoried in caTissue Core, a caBIG tool for biospecimen repositories. Tissues are distributed to WCI investigators with IRBapproved protocols to further the clinical, translational, basic and epidemiologic research. Pathology services of the Core are provided by the Research Pathology Laboratory, a full service lab located on the 5th floor of the WCI building with experienced personnel and equipment for tissue processing, histological and immunohistochemical staining and analysis, laser capture microscopy, tissue microarrays and high throughput, automated image analysis for immunofluorescent and bright field microscopy. This lab is capable of processing high volumes of tissue samples derived from human disease and animal models and works in close collaboration with the other Cores and scientific programs of the WCI as demonstrated by research productivity and support of group science grants. The success of this Core is also highlighted by its contract with the NCI for collecting and processing glioblastoma specimens for The Cancer Genome Atlas Project. Together, the HTPS and the Research Pathology Lab provide centralized, efficent and high quality services to WCI investigators for acceleration of scientific discovery
Keywords: Acceleration; Alcohols; Anatomy; Animal Model; Atlases; base; Basic Science; Biopsy Specimen; cancer Biomedical Informatics Grid; Cancer Center Support Grant; cancer genome; Categories; Clinical; Clinical Research; Clinical trial protocol document; Collaborations; Communications Personnel; Consent; Contracts; Core Facility; Diagnostic; Epidemiologic Studies; Equipment; Equipment and supply inventories; Excision; experience; Floor; Formalin; Freezing; Funding; Glioblastoma; Grant; Harvest; Health Services Research; Hospitals; Hour; Human; human disease; Human Pathology; Human Resources; human tissue; Image Analysis; Individual; Institutes; Investigation; Laboratories; Lasers; Liquid substance; Malignant Neoplasms; Medicine; member; Microscopy; Nitrogen; Operating Rooms; Operative Surgical Procedures; Pathologist; Pathology; Patient Care; Preparation; Process; Productivity; programs; prospective; Protocols documentation; quality assurance; repository; Research; Research Ethics Committees; Research Personnel; Research Support; Resource Sharing; RNA; Schedule; Science; Scientist; Services; Site; Solutions; Source; Specimen; Staining method; Stains; success; Support Groups; Surgical Pathology; Tissue Banking; Tissue Banks; Tissue Microarray; tissue processing; Tissue Procurements; Tissue Sample; Tissues; tool; Training; Translational Research; tumor; Tumor Bank; University Hospitals; Work
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5P30CA138292-03_5776 (2011): $169238
STRUCTURAL STUDIES OF GROWTH FACTOR RECEPTOR FUNCTION
J Daniel, Professor
Johns Hopkins Universitycity: Baltimore country: United States (us)
Grant 9R01GM099321-10 from National Institute Of General Medical Sciences
Abstract: The epidermal growth factor receptor (EGFR/ErbB) family of receptor tyrosine kinases (RTKs) includes four family members in humans EGFR (HER1, ErbB1), HER2 (ErbB2, Neu), HER3 (ErbB3), and HER4 (ErbB4). Each family member consists of an extracellular ligand-binding region, a single membrane-spanning region, a cytoplasmic tyrosine kinase, and a C-terminal tail of ~230 amino acids. All EGFR/ErbB family members are essential for normal embryonic development, and abnormal activity of each ErbB has been associated with human cancer. In particular, overexpressed or active EGFR and HER2 are associated increased severity of lung, colon, and head-and-neck cancers (EGFR) or breast cancer (HER2), and several drugs targeting EGFR and HER2 are FDA-approved anticancer therapies. The canonical model of vertebrate EGFR activity is that ligand binding to the extracellular region stabilizes a specific dimeric conformation of the receptor that in turn drives the kinase domain into an asymmetric dimer in which its activity is stimulated. The activated kinase then phosphorylates several sites in the receptor C-tail as well as other substrates, which results in changes in the localization and/or activity of downstream effectors and initiation of signaling cascades that alter cell growth and differentiation. Much has been learned about the mechanisms governing ErbB activity from X-ray structural studies of ErbB fragments, which have also had a large impact on the design and understanding of ErbB-targeted therapeutics, but several key questions remain. For example, differences in the structure and behavior of Drosophila and vertebrate EGFRs appeared to suggest fundamentally different activation mechanisms for these receptors including the presence of a 12 ligandEGFR signaling complex in Drosophila. A vertebrate 12 ligandEGFR complex seemed ruled out by the symmetric 22 ligandEGFR complexes observed in crystals of vertebrate EGFRligand complexes and the presence of an autoinhibited (and apparently dimerization incompetent) conformation of vertebrate ErbBs in the absence of ligand. Recent results from my laboratory suggest that a 12 ligandEGFR complex also exists for vertebrate ErbBs, however, and we propose (i) to confirm the existence of this 12 complex and probe its structure using both X-ray structural and cell-based functional studies. The presence of a 12 ligandreceptor complex for vertebrate ErbBs would resolve what seemed to be different behavior of Drosophila and vertebrate ErbB extracellular regions, and we propose (ii) to investigate whether the Drosophila kinase regions are governed by a vertebrate-like "asymmetric dimer" mechanism. The ErbB-related type I insulin-like growth factor receptor also mediates key cellular processes, and we propose (iii) to initiate structural studies of this receptor to understand how ligand binding regulates its activity and its relationship to ErbB mechanisms. These studies will establish a molecular basis for understanding of the physiological properties of these key receptors and illuminate new or optimal strategies to target their function in disease states. The epidermal growth factor receptor (EGFR/ErbB) family of receptor tyrosine kinases comprises four members in humans, each of which is essential for normal development and has been associated with human cancers. Abnormal activity of family members EGFR and HER2 in particular are associated with increased cancer severity and is the target of several FDA-approved anticancer drugs. By studying the molecular mechanisms governing the activity of these and related receptors we will gain important insight into how these receptors mediate their normal biological activities, how regulation of their activity breaks down in disease, and what molecular strategies are likely to best inhibit activated forms of these receptors
Keywords: Adopted; Amino Acids; Antibodies; Antineoplastic Agents; base; Behavior; Biological; Biological Assay; Breast; C-terminal; cancer therapy; Cell Differentiation process; cell growth; Cell physiology; Cell Proliferation; Cell Surface Receptors; Cells; Colon; Complex; Coupled; Couples; Curiosities; Cysteine; design; Development; dimer; Dimerization; Disease; DNA Sequence Rearrangement; Drosophila genus; Drug Delivery Systems; drug mechanism; Embryonic Development; Epidermal Growth Factor; Epidermal Growth Factor Receptor; ErbB Receptor Family Protein; ERBB2 gene; ERBB3 gene; ErbB4 gene; Event; extracellular; Extracellular Structure; falls; Family; Family member; FDA approved; Fibroblast Growth Factor; Funding; glycosylation; Goals; Growth Factor Receptors; Head and Neck Cancer; Human; human disease; IGF1R gene; impression; inhibitor/antagonist; insight; Insulin Receptor; Insulin-Like Growth Factor Receptor; interest; Laboratories; Learning; Ligand Binding; Ligands; Lung; malignant breast neoplasm; Malignant Neoplasms; Mammalian Cell; Mediating; member; Membrane; Metabolic; migration; Modeling; Molecular; Molecular Conformation; Mutagenesis; Nature; Nerve Growth Factors; Oncogenic; overexpression; Pharmaceutical Preparations; Phosphorylation Site; Phosphotransferases; Physiological; Property; Protein Tyrosine Kinase; receptor; receptor function; Receptor Protein-Tyrosine Kinases; Regulation; Resolution; Roentgen Rays; Severities; SHFM1 gene; Signal Transduction; Site; stem; Structure; Tail; Testing; Therapeutic Monoclonal Antibodies; therapeutic target; Type I Insulin; Tyrosine Kinase Domain; Vascular Endothelial Growth Factors; Work; X-Ray Crystallography
Relevance: The epidermal growth factor receptor (EGFR/ErbB) family of receptor tyrosine kinases comprises four members in humans, each of which is essential for normal development and has been associated with human cancers. Abnormal activity of family members EGFR and HER2 in particular are associated with increased cancer severity and is the target of several FDA-approved anticancer drugs. By studying the molecular mechanisms governing the activity of these and related receptors we will gain important insight into how these receptors mediate their normal biological activities, how regulation of their activity breaks down in disease, and what molecular strategies are likely to best inhibit activated forms of these receptors
Project start date: 2001-04-01
Project end date: 2015-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-10-067
9R01GM099321-10 (2011): $311600
ORGANIZATION AND FUNCTION OF CELLULAR STRUCTURE
J Daniel, Professor
Duke Universitycity: Durham country: United States (us)
Grant 5T32GM007184-37 from National Institute Of General Medical Sciences
Abstract: The Duke University Program in Cell and Molecular Biology (CMB) provides an entry portal for PhD training in biological sciences, with the objective of training students for careers in science-related professions. CMB provides an interdisciplinary core curriculum that exposes students to diverse topics before selecting their final PhD program. Students select the topics of greatest interest in a modular core class format that reduces class size to maximize interaction with faculty instructors. Teaching is largely based on critical reading of primary literature, supplemented by training in various quantitative skills and coaching in the design and presentation of research proposals. The core course is complemented by elective courses in many areas of concentration. The program features a laboratory rotation system that allows students to participate in the research in each of three well-equipped laboratories of their choice before selecting an advisor. Students may apply and be admitted directly to the University Program in Cell and Molecular Biology. Prior to the second year of study at Duke, students choose the program in which they will earn the Ph.D, from among the following Biochemistry, Biology, Cell Biology, Computational Biology and Bioinformatics, Genetics and Genomics, Immunology, Molecular Cancer Biology, Molecular Genetics and Microbiology, Neurobiology, Pathology, or Pharmacology. RELEVANCE Basic scientific research is the engine of discovery that produces new breakthroughs. This program trains students to perform cutting-edge research in several medically relevant areas
Keywords: Cellular Structures
Project start date: 1975-07-01
Project end date: 2015-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5T32GM007184-37 (2011): $795384
DATA MANAGEMET AND STATISTICAL CORE
J Daniel, Assistant Professor
Case Western Reserve Universitycity: Cleveland country: United States (us)
Abstract: The Data Management and Biostatistics Core will build the physical infrastructure, software solutions and human resources for a centralized, web-based fully GCP-compliant data management system and a program-wide data warehouse and provide support for the analyses of complex longitudinal data systems. The Core will serve the data storage and analytic needs of the 3 Research Projects conducted in all malaria endemic sites ofthe ICEMR and interact directly with the Molecular Diagnostics Core that will create an archive of biological samples and perform diagnostic tests related to malaria infections of humans and mosquitoes. The Specific Aims are to 1 Set up a robust and comprehensive, centralized, web-based fully GCP-compliant data management system for ICEMR program 2; Create a collaborative environment between Papua New Guinea and participating sites in Solomon Islands and other ICEMR institutions and countries for the design, development, testing, and implementation of the data management system 3 Establish a centralized data repository that provides secure access to researchers and collaborators 4 Develop adequate data analysis tools for the analyses of complex longitudinal datasets (with multiple endpoints) 5 Analyze data and prepare results in a timely manner for interim analyses, publications, and presentations 6; Ensure regulatory compliance of data collection management 7 Provide training and development opportunities to Papua New Guinea and Solomon Islands in the areas of epidemiology, biostatistics, and database architecture The core will be directed by Prof. John Aponte from CRESIB, where he has organized similar data systems for large scale malaria epidemiology studies and clinical trials in Sub-Saharan Africa, and supported by Dr. Ivo Mueller, leader ofthe Epidemiology Project, and Thomas Adiguma, senior data manager at PNGIMR. Infrastructure and capacity building will be accomplished by the provision of training and resources in the fields of epidemiology, biostatistics, and dataset architecture
Keywords: Africa South of the Sahara; Agreement; Architecture; Archives; Area; Asia; base; Biological; Biometry; Biostatistics Core; Clinical Research; Clinical Trials; cohort; Collaborations; Collection; Complex; Computer software; Confidentiality; Country; Culicidae; Data; Data Analyses; Data Base Management; Data Collection; Data Coordinating Center; data integrity; data management; Data Set; Data Storage and Retrieval; Databases; design; Development; Diagnostic; Diagnostic tests; Educational workshop; Ensure; Environment; Epidemiology; epidemiology study; Ethics; follow-up; Future; Goals; Good Clinical Practice; Human; Human Resources; human subject; Individual; Infection; Information Systems; Institution; Laws; Malaria; Modeling; Molecular; Online Systems; Papua New Guinea; Participant; Plasmodium; Pregnancy; Privacy; Procedures; programs; Province; Publications; Regulation; relational database; Reporting; repository; Research; Research Ethics Committees; Research Infrastructure; Research Personnel; Research Project Grants; Resources; Risk; Sampling; Secure; sharing data; Site; Solomon Islands; Solutions; System; Testing; TimeLine; tool; Training; transmission process; Vivax Malaria
Relevance: Sophisticated relational database management, compliance with Good Clinical Practice and advanced biostatistical analysis are all essential to the conduct of ethical research related to the control and elimination of malaria. The Data Management and Biostatistics Core will serve this essential function forthe ICEMR
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5U19AI089686-02_6401 (2011): $112455
CANCER AND LEUKEMIA GROUP B STATISTICAL CENTER
J Daniel, Director, Cancer Center Statistics
Mayo Cliniccity: Rochester country: United States (us)
Grant 5U10CA033601-33 from National Cancer Institute
Abstract: The Cancer and Leukemia Group B (CALGB) Statistical Center has primary responsibility for all statistical, data management, computing, information systems, and related activities for the CALGB. Faculty and staff statisticians and data coordinators collaborate closely with study chairs, committee chairs, and other investigators on the design, conduct, analysis, and reporting of all clinical trials and other studies conducted by the CALGB. The Statistical Center has responsibility for central data quality control and database administration. All clinical data are sent to the Statistical Center for editing, verification, and entry into the official CALGB database. Scientific laboratory data are transmitted electronically and become part of the official record. Statisticians and data coordinators from the Statistical Center are assigned to each CALGB study. They work closely with study chairs to monitor the studies and to prepare all necessary reports, analyses, and manuscripts
Keywords: Cancer and Leukemia Group B; Clinical Data; Clinical Trials; Data; data management; Data Quality; Databases; design; Faculty; Information Systems; Laboratories; Manuscripts; Monitor; Quality Control; Reporting; Research Personnel; statistical center; Work
Project start date: 1982-12-01
Project end date: 2015-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
5U10CA033601-33 (2011): $4435349
Sponsored Links Excellgen http://Excellgen.com
CALCIUM SIGNALING IN THE PUBERTAL ACTIVATION OF THE GNRH PULSE GENERATOR
J Daniel, Associate Research Scientist
Yale Universitycity: New Haven country: United States (us)
Grant 5R01HD053529-06 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: The broad, long-term objective is to elucidate the mechanism(s) of the pubertal increase in gonadotropin- releasing hormone (GnRH) secretion required for sexual maturation and fertility. The specific aim is to test the hypothesis that pubertal changes in calcium (Ca2+) and calcium-activated potassium (K(Ca)) channel activity in GnRH neurons underlie the pubertal increase in GnRH secretion. The project will be the first to investigate pubertal changes in Ca2+ and K(Ca) channel activity in GnRH neurons in brain slices and to correlate the changes with GnRH secretion. The knowledge obtained may aid in developing new leads for contraception, as well as treatments for infertility and the pubertal disorders polycystic ovary syndrome, hypogonadism, precocious puberty, and delayed puberty, which possibly result from an abnormal increase in GnRH secretion. These public health goals are relevant to the mission of the Reproductive Sciences Branch of the National Institute for Child Health and Human Development. The project´s rationale is that since neuroendocrine secretion depends on increased cytoplasmic free Ca2+ ([Ca2+]j), the mechanism(s) of the pubertal increase in GnRH secretion likely involves a pubertal increase in [Ca2+]j. This may be due to Ca2+ entry through voltage-dependent Ca2+ channels or Ca2+ release from intracellular stores in response to activation of neurotransmitter and/or hormone receptors on GnRH neurons by presynaptic neurons that convey information about age, growth, availability of metabolic fuels such as glucose and fats, circadian rhythm, and other factors. Ca2+ and K(Ca) currents of green fluorescent protein (GFP)-labeled GnRH neurons will be recorded using the perforated-patch technique, characterized by subtype with pharmacological antagonists, and compared in brain slices of gonadectomized and testosterone-clamped, as well as of gonadal-intact, male prepubertal, pubertal, and adult GnRH-GFP transgenic mice. GnRH in slice superfusates will be measured using radioimmunoassay. The relative contributions of the Ca2+ channel subtypes to increased [Ca2+]j will be assessed by comparing K(Ca) channel activity and GnRH secretion in the absence and presence of Ca2+ channel subtype antagonists. The contribution of Ca2+ mobilization to increased [Ca2+]j will be determined by measuring K(Ca) channel activity in response to kisspeptin activation of G protein-coupled receptor 54. There may be pubertal changes in the activity of one or more of the Ca2+ and K(Ca) channel subtypes, e.g., an increase in the activity of L-type Ca2+ channels, which are important for secretion in other endocrine cells, or increases in the activity of N-, P/Q-, and/or R-type Ca2+ channels, which are important for secretion in other neurons. It is also expected that kisspeptin-induced Ca2+ mobilization will increase at puberty and that these changes will account for the pubertal increase in GnRH secretion
Keywords: Accounting; Adult; Age; Agonist; base; Brain; Calcium; Calcium Signaling; Cells; Child; Child health care; Circadian Rhythms; Contraceptive methods; Delayed Puberty; Disease; Endocrine; Fatty acid glycerol esters; Fertility; G Protein-Coupled Receptor 54; Glucose; Goals; Gonadotropin Hormone Releasing Hormone; Green Fluorescent Proteins; Growth; Hormone Receptor; Human Development; Hypogonadism; Infertility; Institutes; kisspeptin; Knowledge; Label; male; Measures; Metabolic; Mission; Mus; Neurons; Neurosecretory Systems; Neurotransmitters; Physiologic pulse; Polycystic Ovary Syndrome; Potassium; Precocious Puberty; prepuberty; presynaptic; programs; Puberty; public health medicine (field); Radioimmunoassay; Relative (related person); reproductive; Research; Research Personnel; response; Role; Science; Sexual Maturation; Slice; Techniques; Testing; Testosterone; Transgenic Mice; voltage
Project start date: 2007-04-01
Project end date: 2012-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PA-03-079
5R01HD053529-06 (2011): $174794
J Daniel, Director, Cancer Center Statistics
Mayo Cliniccity: Rochester country: United States (us)
Abstract: The Biostatistics Shared Resource (BSR) of the Mayo Clinic Cancer Center (MCCC) has provided expert statistical collaboration with MCCC investigators for over 35 years. The purpose is to provide statistical and biomathematical collaboration for the development and conduct of peer-reviewed cancer research generated by MCCC investigators. Research encompasses basic science, clinical trials, epidemiologic research, and other translational and educational research. The primary usage of CCSG funds is to support statistical and biomathematical collaboration on pilot projects, for assistance to investigators in developing approved research projects not otherwise funded but leading towards external funding, and to support unanticipated needs for statistical collaboration on MCCC approved projects. Over its history the BSR has undergone a natural and deliberate evolution and expansion, spurred by the expanding research activities of the MCCC. In the current grant period the BSR has successfully and strategically expanded in two primary areas high dimensional genomic and proteomic data analysis (4 new faculty with this expertise), and to all 3 MCCC campuses. BSR faculty are physically located on all 3 MCCC campuses, and the entire resource communicates regularly and effectively through multiple mechanisms to ensure coordinated collaboration. The BSR has multiple focus areas (teams) tied together into a well-organized, efficient core. These are (I) a Clinical Trials team responsible for cancer clinical trials, associated translational research, interventional psychosocial research, and patient and public education research projects, (II) a Population Science team responsible for providing statistical collaboration and data management support for cancer observational studies, including genetic and molecular epidemiology, (III) a Quality of Life team responsible for providing general and specific collaboration, measurement tools, and analysis for MCCC investigators investigating the impact of clinical and psychosocial interventions on cancer patients, families, caregivers, and others, and (IV) a Biomathematics team that provides collaborative expertise in the mathematical aspects of imaging, data processing and analysis, mathematical modeling of diease processes, and general mathematical or algorithmic issues. The BSR provides statistical and biomathematical collaboration to MCCC investigators across all three MCCC campuses. The resource has been remarkably productive, with 528 peer reviewed publications in the current grant period, and has collaborated on 104 NIH or NIH equivalent grants in the current grant period
Keywords: Address; Adverse event; American Association of Cancer Research; American Society of Clinical Oncology; American Society of Hematology; anticancer research; Area; Basic Science; Bibliography; biomathematics; Biostatistics Shared Resource; Cancer Center Support Grant; Cancer Patient; Case-Control Studies; Characteristics; Clinical; Clinical Data Update System; Clinical Research; Clinical Trials; Clinical Trials Data Monitoring Committees; cohort; Cohort Studies; Collaborations; Collection; Complex; Comprehensive Cancer Center; computerized data processing; Data Analyses; Data Collection; Data Element; data management; Data Quality; Data Set; Databases; design; Development; Disease; Doctor of Philosophy; Elements; Eligibility Determination; Ensure; Epidemiologic Studies; Epidemiologist; Evolution; Faculty; Family; Family Caregiver; Funding; genetic epidemiology; Genomics; Goals; Grant; Housing; Human Resources; Image; Individual; interest; Intervention; Laboratories; Lead; Learning; Life; Link; Malignant Neoplasms; Manuscripts; mathematical model; Mayo Clinic Cancer Center; Measurement; meetings; member; Molecular Epidemiology; Monitor; Observational Study; Outcome; Patient Education; Peer Review; Phase; Phase III Clinical Trials; Physicians; Pilot Projects; Policies; population based; Population Sciences; Principal Investigator; Procedures; Process; programs; Proteomics; protocol violation; Protocols documentation; psychosocial; public education; Publications; Quality Control; Quality of life; Recording of previous events; Reporting; repository; Research; Research Activity; Research Personnel; Research Project Grants; research study; Resolution; Resource Sharing; Resources; response; Sample Size; Sampling; Scientist; Services; SKIL gene; sound; Specialist; Specimen; Statistical Methods; Surveys; System; Time; tool; Translational Research; United States National Institutes of Health; web site; Work; Writing
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
5P30CA015083-37_5819 (2011): $531906
J Daniel, Prof Of Pathology & Laboratory Medicine
Emory Universitycity: Atlanta country: United States (us)
Abstract: The Animal Models and Pathology Specialized Resource (Core D) of the Emory Molecular and Translational Imaging Center (EMTIC) will assist the overall project by providing service in two critical areas animal models and histopathologic services. The Core is necessary for the success of the EMTIC and is highly integrative, being utilized by all four research projects and all five pilot projects. The animal models component will act to generate and characterize novel rodent models for molecular imaging in cancer research and will assist the individual projects by collecting and analyzing tumor data from these animals. The production and analysis of relevant animal models of cancer is central to the EMTIC because of the heavy emphasis on the development and validation of novel tracers in preclinical studies that could be utilized as markers for tumor detection, progression, and dissemination in vivo. The animal component will also serve as a repository for the preservation and distribution of any novel transgenic strains of mice that may be utilized in the Research Projects and Pilot Projects as they become necessary. The Core will perform necropsy and tissue collection for experimental animals, and will collect tumor samples from selected animals for generation of cell lines for use by EMTIC investigators. The pathology component of the Core will provide tissue and tumor procurement expertise, tissue processing, as well as histological and immunohistochemical analysis of tumor samples from human disease and animal models. Histologic and immunohistochemical evaluation of tumors from patients and animal models will be necessary in order to correlate and validate the detection of novel tracers by PET, MRI and optical methods with the targeted biomarkers in tissues. Thus the Animal Models and Pathology Core will contribute significantly to the overall goals of the EMTIC from a scientific perspective by providing these services as well as expertise. In addition, the inclusion of these services into a comprehensive core component will provide additional benefit to EMTIC as a whole, through consolidation of effort, avoidance of unnecessary experimental duplication, and by drawing upon the collective expertise of the Core personnel
Keywords: Animal Cancer Model; animal data; Animal Model; animal tissue; Animals; anticancer research; Area; Autopsy; Biological Preservation; biomarker; Cancer Model; Cell Line; Consultations; Cryopreservation; Data; Detection; Development; Embryo; Evaluation; Generations; Genetic; Goals; Histologic; Human; human disease; Human Resources; human tissue; Image; in vivo; Individual; Investigation; laser capture microdissection; Magnetic Resonance Imaging; Maintenance; Malignant Neoplasms; Modeling; Molecular; molecular imaging; mouse model; Mouse Strains; Mus; novel; Optical Methods; Pathology; Patients; Pilot Projects; Positron-Emission Tomography; preclinical study; Process; Production; Rattus; Reagent; repository; Research Personnel; Research Project Grants; research study; Resources; Rodent Model; Sampling; Services; success; Technical Expertise; Tissue Banks; tissue processing; Tissue Sample; Tissues; Tracer; Transgenic Organisms; Transplantation; tumor; Tumor Markers; Tumor Tissue; Validation
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
5P50CA128301-04_9004 (2011): $70928
VARIATIONS IN LIPID HOMEOSTASIS AND SIGNALING AFFECT ALPHA-SYNUCLEIN PATHOBIOLOGY
J Daniel, Postdoctoral Associate
Whitehead Institute For Biomedical Rescity: Cambridge country: United States (us)
Grant 5F32NS066692-02 from National Institute Of Neurological Disorders And Stroke
Abstract: Synucleinopathies are a class of neurodegenerative disease characterized by pathological lesions composed of aggregates of a-synuclein (a-syn) in select populations of neurons. Alpha-synuclein is a small, abundant lipid-binding protein of unknown function that is prone to misfolding when not bound to lipids. The accumulation of a-syn aggregates in synucleinopathies suggests that aberrations in protein homeostasis might contribute to pathogenesis in these diseases. Because protein homeostasis mechanisms are highly conserved throughout eukaryotes, yeast cells can be used as a tractable model organism to investigate factors that contribute to the pathogenesis of synucleinopathies. The Lindquist laboratory has developed a yeast model over-expressing human a-syn that recapitulates many aspects of a-syn toxicity, and has observed that sterol biosynthesis and trafficking are also perturbed. Disrupting sterol biosynthesis can potentially alter a variety of cellular pathways. In fact, sphingolipid production is responsive to flux through the sterol pathway. Interestingly, both pathways have been implicated independently in a-syn toxicity and disease progression. The contribution of lipids to a-syn toxicity remains unresolved, and understanding how lipids affect synucleinopathy pathogenesis is essential for identifying potential therapeutic targets. The first aim of my proposal will investigate how the yeast lipidome is altered in response to a-syn expression. Lipidomic profiling has performed on yeast expressing nontoxic and toxic levels of a-syn in collaboration with Dr. Clary Clish at the Broad Institute. The lipid species identified up to this point correlate with those predicted to change upon a-syn expression. These results will then be compared to the lipid profiles of yeast co-expressing a-syn and genetic modifiers we discovered using overexpression and deletion screens. This will identify the lipids that change when toxicity is suppressed or enhanced. These targets lipids will then be manipulated in our yeast a-syn model to determine their effect on toxicity. My second aim is to evaluate whether the defects in cellular unesterified ergosterol distribution caused by a-syn expression lead to the mis-trafficking and accumulation of sphingolipids. Fluorescent microscopy will be used to monitor sphingolipid trafficking and localization in yeast cells. In addition, sphingolipids will be quantified using metabolic labeling. The cellular localization of unesterified ergosterol will be determined using filipin stain. Finally, my last aim will be evaluate sterol-sphingolipid regulation defects in cultured DA neurons expressing a-syn. Similar to aim 2, fluorescent microscopy will be used to monitor the trafficking and localization of unesterified cholesterol and sphingolipids, respectively. The amount of sphingolipids and sterols in a-syn expressing DA neurons will be quantified as well. Lipids are important macromolecules that are involved in many cellular events and have been implicated in several human diseases. Investigation of their contribution to neurodegenerative disease specifically PD, will promote further understanding of lipid homeostasis and a-syn pathobiology. The identification of lipids that might affect a-syn toxicity will be extremely useful in developing lipid-specific therapeutics reducing the pleiotropic effects associated with the general lipid drugs currently employed
Relevance: Lipids are important macromolecules that are involved in many cellular events and have been implicated in several human diseases. Investigation of their contribution to neurodegenerative disease specifically PD, will promote further understanding of lipid homeostasis and ¿-syn pathobiology. The identification of lipids that might affect ¿-syn toxicity will be extremely useful in developing lipid-specific therapeutics reducing the pleiotropic effects associated with the general lipid drugs currently employed
Project start date: 2010-09-01
Project end date: 2013-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-09-210
5F32NS066692-02 (2011): $51326
ACTIVITY-DEPENDENT REGULATION OF NEUROMUSCULAR JUNCTION FORMATION
J Daniel, Professor/research Professor
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Grant 5R01NS059848-03 from National Institute Of Neurological Disorders And Stroke
Abstract: Synaptogenesis is regulated by both activity-dependent and independent mechanisms. Activity-dependent regulation of synaptogenesis is mediated, in part, by changes in gene expression. Therefore an important goal of neuroscientists is to understand the mechanisms by which synaptic activity is transduced to the genome to influence synapse formation. The neuromuscular junction (NMJ) is a well studied model for identifying and characterizing the mechanisms by which synaptic activity influences synapse formation and regulates gene expression. Although activity-dependent control of NMJ formation has been studied for decades, the underlying molecular mechanisms are poorly understood. We recently identified a Dach2-dependent signal transduction cascade that contributes to activity-dependent expression of nAChR and MuSK genes. Therefore, this signaling cascade along with the previously reported HDAC9 (MITR) signaling cascade, that also participates in activity-dependent nAChR gene expression, represent good candidates for mediating the effects of synaptic transmission on NMJ formation. In this grant application we propose to use a combination of genetic approaches and cell and molecular biological approaches to investigate the role Dach and MITR signaling play in regulating NMJ formation and controlling gene expression by nerve-induced muscle depolarization. Knockout animals will be used to study NMJ development, while innervated and denervated skeletal muscle will be used to study nerve-induced, activity-dependent gene regulation. Specifically, we propose to 1) Evaluate the role Dach2 and MITR play during development of the NMJ; 2) Examine if HDAC4 coordinates Dach2 and MITR gene repression in response to muscle innervation; and 3) Determine if the Dach interacting proteins Six and Eya participate in activity-dependent regulation of gene expression. This research will identify mechanisms by which muscle activity regulates synapse formation during development and modifies synapse and muscle function in the adult. Although this research is of a basic nature, it may suggest ways of enhancing synapse formation, synaptic plasticity and muscle function in the injured, diseased or aged individual. The studies in this grant application aim to understand how muscle activity signals to the genome to regulate neuromuscular development, muscle atrophy and muscle gene expression. These studies will not only further our understanding of the mechanisms underlying these events, but may also suggest novel strategies for restoring neuromuscular communication and improving muscle function following injury or disease
Keywords: Adult; aged; Applications Grants; Binding Sites; Biological; Biological Assay; Cells; Choline O-Acetyltransferase; Communication; Complex; Data; Development; Disease; DNA Binding; Elements; Event; Exhibits; Family member; follow-up; Gene Expression; Gene Expression Regulation; gene induction; gene repression; Genes; Genetic; Genetic Transcription; Genome; Goals; Grant; HDAC4 gene; Health; Histone Deacetylase; improved; in vivo; Indium; Individual; injured; Injury; interest; knockout animal; Knockout Mice; Mediating; Molecular; Muscle; Muscle denervation procedure; Muscle function; Muscle-Specific Kinase; Muscular Atrophy; muscular structure; Mutant Strains Mice; Myogenin; Nature; Nerve; nerve supply; neuromuscular; Neuromuscular Junction; neurotransmission; Nicotinic Receptors; novel strategies; Phosphoric Monoester Hydrolases; Play; postsynaptic; presynaptic; Promotor (Genetics); Protein Binding; Proteins; Regulation; Reporting; Repression; Research; response; Role; Signal Transduction; Skeletal muscle structure; Small Interfering RNA; Staging; Study models; Synapses; synaptic function; Synaptic plasticity; Synaptic Transmission; synaptogenesis; Testing; Transcription Coactivator; transcription factor; Variant
Relevance: The studies in this grant application aim to understand how muscle activity signals to the genome to regulate neuromuscular development, muscle atrophy and muscle gene expression. These studies will not only further our understanding of the mechanisms underlying these events, but may also suggest novel strategies for restoring neuromuscular communication and improving muscle function following injury or disease
Project start date: 2009-05-15
Project end date: 2013-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-070
5R01NS059848-03 (2011): $323790
STEM CELLS AND CELL THERAPIES IN LUNG BIOLOGY AND DISEASES
J Daniel, Associate Professor
University Of Vermont & St Agric Collegecity: Burlington country: United States (us)
Grant 2R13HL097533-02 from National Heart, Lung, And Blood Institute
Abstract: Stem Cells and Cell Therapies in Lung Biology and Diseases Repair and regeneration of diseased lung with stem cells is an exciting potential therapeutic approach for a variety of lung diseases. Increasing information suggests that cells normally not resident in the lung can be utilized to impair lung injury and also to induce repair and remodeling after injury. In parallel, there has been a surge in bioengineering studies investigating use of artificial and de-cellularized lung matrices as framework for three-dimensional lung regeneration. The combination of these studies with those utilizing stem cells and/or cell therapies is a promising rapidly developing direction but one that is still in its infancy. These studies have been further paralleled by significant increases in understanding the molecular and cellular events by which stem and/or progenitor cells resident in the lung participate in both lung development and in repair and remodeling after lung injury. We have held three previous conferences at the University of Vermont in July 2005, "Adult Stem Cells, Lung Biology, and Lung Disease", sponsored by the NHBLI and Cystic Fibrosis Foundation; July 2007 "Stem Cells and Cell Therapies in Lung Biology and Disease", sponsored by the NHLBI through an R13 conference grant as well as by the Alpha 1 Antitrypsin Foundation and Pulmonary Fibrosis Foundation; and July 2009 "Stem Cells and Cell Therapies in Lung Biology and Disease", sponsored by the NHLBI through an R13 conference grant as well as by the Alpha 1 Antitrypsin Foundation, Emory Center for Respiratory Health, LAM Treatment Alliance, and Pulmonary Fibrosis Foundation that brought together relevant investigators as well as interested junior faculty and trainees to debate and discuss issues in this rapidly moving field. These conferences have been very successful and have both stimulated the field and resulted in a series of guidelines for basic and translation research to be utilized by both investigators and by funding agencies. As studies of stem cell and cell therapies for lung diseases continues to move at a rapid pace, we propose to again convene the relevant investigators as well as representatives from the NHLBI, FDA, and non-profit Respiratory Disease Foundations to again debate and discuss current issues. One area in particular to be discussed is the balance between basic and translational research and the role of clinical trials, particularly as there have now been clinical trials of stem cells for pulmonary hypertension and for COPD. As in the previous conferences, junior investigators and trainees (graduate, post-doctoral, fellow) will be particularly targeted for inclusion in both conference discussions as well as in poster sessions. As a continuing feature, trainees will be able to compete for an increased number of travel awards based on blinded review of poster s. The conference is planned for July 2011 at the University of Vermont. Enthusiasm is already high among potential participants. We anticipate that this conference will again foster extensive discussion and debate and will significantly guide the directions taken in basic and clinical research of stem cells and cell therapies for lung diseases
Keywords: ing; Address; adult stem cell; alpha 1-Antitrypsin; American; Area; Award; base; Basic Science; Biology; Biomedical Engineering; Blinded; Cell Therapy; Cells; Chest; China; Chronic Obstructive Airway Disease; Clinical; Clinical Research; Clinical Trials; Communities; Cystic Fibrosis; Data; design; Development; Disease; embryonic stem cell; Equilibrium; Ethics; Event; Faculty; Fostering; Foundations; Funding Agency; Funding Opportunities; Future; Goals; graduate student; Grant; Guidelines; Health; induced pluripotent stem cell; infancy; Injury; insight; interest; International; Lung; lung development; Lung diseases; lung injury; Malignant neoplasm of lung; meetings; Molecular; Motivation; National Heart, Lung, and Blood Institute; Natural regeneration; North America; Participant; phrases; Postdoctoral Fellow; posters; Publishing; Pulmonary Fibrosis; Pulmonary Hypertension; Recommendation; repaired; Research; Research Personnel; Research Support; Respiratory Center; Role; Series; Societies; stem; Stem cells; symposium; Therapeutic; Therapeutic Studies; Time; Translational Research; translational study; Travel; United States National Institutes of Health; Universities; Vermont
Relevance: Stem Cells and Cell Therapies in Lung Biology and Diseases Repair and regeneration of diseased lung with stem cells is a rapidly moving field and an exciting potential therapeutic approach for a variety of lung diseases. We propose a conference in July 2011 on Stem Cells and Cell Therapies in Lung Biology and Diseases. This conference will convene relevant experts along with interested junior investigators and trainees to discuss and debate current issues in the field and to craft guidelines for basic and translational research and for funding agencies supporting this research
Project start date: 2009-07-01
Project end date: 2012-06-30
Budget start date: 15-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-10-071
2R13HL097533-02 (2011): $30099
TRANSCRIPTIONAL REGULATION OF MYOGENIC STEM CELLS
J Daniel, Director/professor
University Of Minnesota Twin Citiescity: Minneapolis country: United States (us)
Grant 5R01AR055906-03 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Somatic stem cell populations reside in adult tissues and participate in the maintenance and regeneration of adult tissues. The signaling networks that govern the somatic stem cell populations are ill defined. We have previously defined the forkhead/winged helix transcription factor, Foxk1, as the first molecular marker for the muscle stem cell population that resides in adult skeletal muscle. Using a gene disruption strategy, mice lacking Foxk1 have impaired muscle regeneration, decreased numbers of muscle stem cells that have perturbed cell cycle kinetics and an induction of the cyclin dependent kinase inhibitor, p21. We provide new preliminary data supporting a novel pathway whereby Foxk1 contributes to a repression complex that represses p21 resulting in the activation and proliferation of the muscle stem cell population. These preliminary data provide a platform for the studies, outlined in this revised grant application. The overall hypothesis of this proposal is that lineage specific transcriptional regulators interact and modulate the myogenic stem cell population to promote skeletal muscle regeneration. To address this hypothesis, we will pursue the following three specific aims Specific Aim #1 To define the transcriptional regulation of the cyclin dependent kinase inhibitor, p21CIP in the muscle stem cell population. Specific Aim #2 To define the transcriptional role of Foxk1 as a critical regulator of the muscle stem cell population. Specific Aim #3 To define the functional role of Foxk1 overexpression in adult skeletal muscle. These studies will enhance our understanding of the transcriptional regulation of the myogenic stem cell population and the mechanisms, in part, that govern muscle regeneration following an injury or disease. Furthermore, the results of these studies will serve as a platform for therapeutic strategies directed towards the treatment of acquired myopathic diseases. Congenital and acquired myopathies are common and deadly. The capacity of adult tissues to regenerate in response to disease or injury is due to resident stem cell populations. The regulatory mechanisms that govern muscle stem cells are unknown. This proposal will decipher the molecular pathways that govern muscle stem cells and may ultimately lead to novel therapies directed towards an enhanced regenerative response in patients with myopathic diseases
Keywords: 21+ years old; Address; Adult; adult human (21+); Aging; Applications Grants; Assay; Binding; Binding (Molecular Function); Bioassay; Biochemical; Biologic Assays; Biological; Biological Assay; Biological Models; biological signal transduction; Body Tissues; CAP20 protein; casein kinase II; Casein Kinase TS; Casein Kinase-2; CDK Inhibitor Protein; CDK-Interacting Protein 1; Cdk2 inhibitor protein; CDK2-associated protein 20 kDa; CDKI Protein; CDKN1 protein; CDKN1A; CDKN1A Protein; cdn1 protein; Cell Communication and Signaling; Cell Cycle Kinetics; Cell Cycle Progression; Cell Kinetics; Cell Signaling; CIP-1 Protein; Cip1 protein; Clinical; Co-Immunoprecipitations; Complex; Cyclin Kinase Inhibitor; Cyclin-Dependent Kinase Inhibitor; Cyclin-Dependent Kinase Inhibitor 1A; cyclin-dependent kinase Inhibitor p21; Data; Deterioration; Disease; disease/disorder; Disorder; Disorder of muscle, unspecified; Emergent Technologies; Emerging Technologies; exhaustion; experiment; Experimental Designs; experimental research; experimental study; Foundations; Gene Down-Regulation; gene repression; Genes; Genetics-Mutagenesis; Grant Proposals; Grants, Applications; Health; heavy metal lead; heavy metal Pb; Human, Adult; Injury; injury response; Intracellular Communication and Signaling; Knockout Mice; Laboratories; Lead; Maintenance; Maintenances; Mammals, Mice; MDA 6; mda-6 protein; Methods and Techniques; Methods, Other; Mice; Mice, Knock-out; Mice, Knockout; Model System; Models, Biologic; Molecular; Molecular Biology, Mutagenesis; Molecular Interaction; molecular marker; Mother Cells; mouse model; Murine; Mus; Muscle; Muscle Disease; Muscle disease or syndrome; Muscle Disorders; muscle regeneration; Muscle satellite cell; muscle stem cell; Muscle Tissue; Muscle, Skeletal; Muscle, Voluntary; Muscular Diseases; muscular disorder; Mutagenesis; mutant; Myopathic Conditions; Myopathic disease or syndrome; Myopathic Diseases and Syndromes; Myopathy; Myopathy, unspecified; Natural regeneration; novel; novel marker; Nuclear; Null Mouse; oncoprotein p21; overexpression; P21; p21 cell cycle regulator; p21 cyclin kinase inhibitor; p21(cip1); p21(waf1-cip1); p21-WAF1; pathway; Pathway interactions; Patients; Pb element; Physiologic; Physiological; Pic-1 protein (cyclin); Population; Progenitor Cells; programs; Programs (PT); Programs [Publication Type]; Protein Kinase CK2; Protein Kinase CKII; protein p21; regenerate; Regeneration; regenerative; Regulation; Repression; Research; research study; Residual; Residual state; response; response to injury; Role; sarcopenia; satellite cell; self-renewal; Senescence; senescent; senescent cell-derived inhibitor protein 1; Signal Transduction; Signal Transduction Systems; Signaling; Skeletal muscle structure; Skeletal Muscle Tissue; social role; stem cell population; Stem cells; Techniques; Technology; Therapeutic; Tissues; Transcription Corepressor; transcription factor; Transcription Regulation; Transcription Repression; Transcription Repressor; Transcription Repressor/Corepressor; Transcriptional Control; Transcriptional Corepressor; Transcriptional Regulation; Transcriptional Repression; Transcriptional Repressor; Transcriptional Repressor/Corepressor; transgenic; Transgenic Mice; Transgenic Organisms; WAF-1 Protein; WAF1 CIP1; WAF1 protein; Winged Helix
Relevance: Congenital and acquired myopathies are common and deadly. The capacity of adult tissues to regenerate in response to disease or injury is due to resident stem cell populations. The regulatory mechanisms that govern muscle stem cells are unknown. This proposal will decipher the molecular pathways that govern muscle stem cells and may ultimately lead to novel therapies directed towards an enhanced regenerative response in patients with myopathic diseases
Project start date: 2009-04-01
Project end date: 2014-02-28
Budget start date: 1-MAR-2011
Budget end date: 29-FEB-2012
PFA/PA: PA-07-070
5R01AR055906-03 (2011): $295990
5R01AR055906-02 (2010): $328878
UBIQUITIN CHAIN EDITING BY THE MAMMALIAN PROTEASOME
J Daniel
Harvard University (medical School)city: Boston country: United States (us)
Grant 1R01GM095526-01A1 from National Institute Of General Medical Sciences
Abstract: Since its discovery, the proteasome has generally been viewed as a passive player in the selection of substrates for degradation. This notion has recently been challenged by studies, many from this laboratory, showing that proteasomes rapidly edit ubiquitin chains and that these processes strongly regulate proteasome output. We first examined these issues in yeast, but now propose to focus on mammals, where little is being done. The regulatory systems differ between yeast and mammals, making it especially important that these questions are studied carefully in mammals. For example, whereas yeast has a single enzyme that trims chains on the proteasome, Ubp6, mammals have two Usp14 (the Ubp6 ortholog) and Uch37. Also, our studies indicate that the influence of chain trimming on degradation rates is more dramatic in mammals. We have developed a specific, small-molecule inhibitor of Usp14. Using this inhibitor, both in vivo and in cells, we discovered an unexpectedly powerful role of chain trimming in regulating degradation. This compound is unique in providing, for the first time, a way to enhance output of the ubiquitin-proteasome pathway. Remarkably, it induces faster degradation of multiple proteotoxic proteins as well as oxidized proteins, and provides resistance to proteotoxic stress. Possible eventual therapeutic applications include, in principle, tens of known diseases in which toxic proteins are expressed or protein degradation is deficient. A screen has also been completed for Uch37, covering 130,000 compounds, and a highly selective inhibitor is in hand. Though Usp14 and Uch37 both trim chains, they seem to regulate the proteasome in distinct ways, for unknown reasons. To understand this, we will first carry out in vitro and model substrate studies testing whether the topological linkage specificity of substrate-bound ubiquitin chains provides an underlying code that could explain the substrate specificities of Usp14 and Uch37 and their effects on degradation rates. Similar model substrate work will assess whether a protein´s susceptibility to unfolding by the proteasome in linked to chain trimming. What features of the substrate determine susceptibility to editing factors will be analyzed further using endogenous substrates? We will use state of the art mass spectrometry approaches to determine, on a broad, if not global, scale, which substrates in the cell are regulated via proteasomal chain editing, using the inhibitors as well and gene knockouts in mice. Relevant properties of endogenous substrates, such as the type of ubiquitin chain they carry and their ubiquitin receptor dependence, will be characterized. A final component of the editing system is Hul5, a conserved, proteasome-associated ubiquitin ligase. We have shown Hul5 extends proteasome-bound ubiquitin chains and is thus important for chain editing in yeast, where it antagonizes Ubp6. Thus far Hul5 has essentially not been studied in mammals. The results will provide important new insights into proteasome function and might have relevance to the development of novel therapeutics. We have discovered a small molecule, and possibly several types of small molecules, that have the unique and unpredicted property of being able to enhance the rate at which certain proteins are destroyed in cells. For the many major diseases in which toxic proteins are expressed, reducing their levels through this new method may have therapeutic benefits. These small molecules inhibit enzymes that normally strip "degradation signals" (ubiquitin groups) from proteins, and therefore they cause such proteins to be eliminated at an exceptionally fast pace
Keywords: Active Sites; Address; Affect; base; Binding (Molecular Function); Biochemical; Biochemistry; Cells; Client; Code; Complement; Dependence; design; Development; Disease; Dissociation; Enzymes; Excision; Funding; Genes; Genetic; Hand; In Vitro; in vitro Model; in vivo; Individual; inhibitor/antagonist; insight; interest; knockout gene; Laboratories; Ligase; Link; Mammals; Mass Spectrum Analysis; Methods; Modeling; multicatalytic endopeptidase complex; Mus; Mutant Strains Mice; novel; novel therapeutics; Orthologous Gene; Output; Pathway interactions; Play; Predisposition; Probability; Process; Property; Proteasome Binding; protein degradation; Proteins; receptor; Regulation; research study; Resistance; response; Role; Signal Transduction; small molecule; Specificity; Stress; Substrate Specificity; System; Testing; Therapeutic; Time; Toxic effect; Ubiquitin; ubiquitin ligase; Work; Yeasts
Relevance: We have discovered a small molecule, and possibly several types of small molecules, that have the unique and unpredicted property of being able to enhance the rate at which certain proteins are destroyed in cells. For the many major diseases in which toxic proteins are expressed, reducing their levels through this new method may have therapeutic benefits. These small molecules inhibit enzymes that normally strip "degradation signals" (ubiquitin groups) from proteins, and therefore they cause such proteins to be eliminated at an exceptionally fast pace
Project start date: 2011-06-01
Project end date: 2015-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-10-067
1R01GM095526-01A1 (2011): $490579
THE ROLE OF CXCL9 IN GENITAL HSV-2 INFECTION
J Daniel
University Of Oklahoma Hlth Sciences Ctrcity: Oklahoma City country: United States (us)
Grant 5R01AI067309-03 from National Institute Of Allergy And Infectious Diseases
Abstract: Sexually transmitted diseases such as herpes simplex virus type 2 (HSV-2) are a significant problem in otherwise healthy women. The number of new cases of genital HSV infection is approaching 500,000 individuals annually with an estimated 40-60 million Americans infected with the virus. HSV-2 infections are particularly severe in immunocompromised individuals and newborns. Demographic characteristics associated with increased HSV-2 prevalence in the USA include female sex, race, lower eductional level, and lower income. Likewise, there does not appear to be a decrease in the incidence of HSV-2 as a result of educational outreach programs even among college students. Based on these results, HSV-2 continues to be a major health problem within the United States. Due to the nature of the infection, a mouse model has been developed in which specific questions related to pathogenic manifestations associated with the infection and the host immune response to the virus can be addressed. We have taken advantage of this model to initiate a study investigating the role of chemokines in the host response to infection. We have found one group of chemokines including monokine induced by interferon-gamma (CXCL9) and interferon gamma-inducible protein 10 (CXCL10) to be expressed in selective tissues post genital HSV-2 infection. Preliminary results have also found mice deficient in the lone receptor for these chemokines, CXCR3, to be highly susceptible to genital infection. We propose to further characterize these initial observations by testing the hypothesis that CXCL9, CXCL10, and CXCR3 are crucial in the control of genital HSV-2 infection by facilitating the recruitment and function of activated T cells and NK cells within the vaginal tissue, spinal cord, and/or brain stem. To test this hypothesis, we plan on using mice deficient in the expression of the chemokine receptor CXCR3, deficient in the ligands CXCL9 or CXCL10, or chimeric and transgenic mice infected with a clinical isolate of HSV-2 to characterize the host immune response to the virus as it relates to virus titer and spread in the infected tissue. In accomplishing this study, it is anticipated we will learn the role of these sentinel chemokines in orchestrating a protective host response to infection and in so doing, facilitate the development of therapeutic strategies (e.g., vaccine or gene transfer) to reduce the incidence of infection or reduce the capacity of latent virus to reactivate
Keywords: 0-11 years old; 0-6 weeks old; Acute; Address; Adenoviridae; Adenoviruses; Adoptive Transfer; American; Animals; Anti-viral Drug Resistance; Anti-viral Drug Resistant; Antiviral Drug Resistance; Antiviral Drug Resistant; base; Body Tissues; Brain Stem; Brainstem; CD183; Cell Function; Cell Locomotion; Cell Migration; cell motility; Cell Movement; Cell physiology; Cell Process; Cells; Cellular Function; Cellular Infiltration; Cellular Migration; Cellular Physiology; Cellular Process; Characteristics; chemoattractant cytokine; chemokine; chemokine receptor; Child; Child Youth; children; Children (0-21); Chimera; Chimera organism; CKR-L2; Clinical; CMK; CMKAR3; college student; Congenic Mice; Congenital herpes simplex; Congenital herpes simplex infection; congenital herpes simplex virus infection; crg-10; CRG-2; CXC chemokine receptor 3; CXCL10; CXCL10 gene; CXCL9; CXCL9 gene; CXCR3; CXCR3 gene; CXCR3 protein; CXCR3 receptor; cytokine; Cytokines, Chemotactic; Cytotoxic cell; Dendritic Cells; Disease; disease/disorder; Disorder; Drug Resistance, Viral; Epidemiologic Research; Epidemiologic Studies; Epidemiological Studies; Epidemiology Research; Female; Gamma interferon; gene product; Gene Transfer; Genital; genital herpes; genital infection; Genital system; Genital System, Female, Vagina; gIP-10; Goals; GPR9; Health; herpes genitalia; Herpes Genitalis; Herpes labialis Virus; herpes simplex ii; Herpes Simplex Virus; Herpes Simplex Virus 2; Herpes Simplex Virus Type 2; Herpes Simplex, Genital; herpesvirus; Herpesvirus 2 (alpha), Human; Herpesvirus 2, Human; Herpesvirus hominis; Herpesvirus progenitalis; HHV-2; Homologous Chemotactic Cytokines; host response; HSV; HSV-2; HSV2; Human; Human (alpha) herpes virus 2; human alphaherpesvirus 2; Human herpes simplex virus type 2; human herpesvirus 1 group; Human Herpesvirus 2; Human, Child; Human, General; Humig; IFI10; IFN-g; IFN-Gamma; IFNG; Immune response; Immunocompromised; Immunocompromised Host; Immunocompromised Patient; immunoresponse; Immunosuppressed Host; immunosuppressed patient; Incidence; Individual; Infant, Newborn; Infection; Infection Control; INP10; Intercrines; Interferon Gamma; Interferon gamma (human lymphocyte protein moiety reduced); Interferon Type II; Interferon, Immune; Interferon-gamma; Investigators; IP-10; IP10; IP10-R; K lymphocyte; Latent Virus; Learning; Leukocyte Trafficking; lFN-Gamma; Ligands; Low income; lymph cell; lymph gland; Lymph node proper; lymph nodes; Lymphocyte; Lymphocytic; Mammals, Mice; Man (Taxonomy); Man, Modern; Medulla Spinalis; Mice; MIG; MIG Gene; Mig-R; MigR; MOB-1; Modeling; Monokine Induced by Gamma Interferon; Mothers; Motility; Motility, Cellular; mouse model; Murine; Mus; Natural Killer Cells; Nature; Neonatal herpes simplex; newborn human (0-6 weeks); Newborn Infant; Newborns; NK Cells; outreach program; pathogen; Predisposition; Prevalence; Production; programs; Programs (PT); Programs [Publication Type]; Proteins; Race; Racial Group; receptor; Receptor Protein; Recurrence; Recurrent; Research Personnel; Researchers; Resistance; resistance to anti-viral; resistance to antiviral; resistant; resistant to anti-viral; resistant to antiviral; Reticuloendothelial System, Lymph Node; Reticuloendothelial System, Spleen; Role; SCYB10; SCYB9; Sentinel; sex; sexually active; Sexually Transmitted Diseases; Sexually Transmitted Disorder; Sexually Transmitted Infection; Simplexvirus; SIS cytokines; Small Inducible Cytokine B9; social role; Spinal Cord; Spleen; STD; Stocks, Racial; Subcellular Process; Susceptibility; T-Cell Activation; T-Cells; T-Lymphocyte; T-Lymphocyte and Natural Killer Cell; T-Lymphocyte and NK-Cell; Testing; therapeutic development; thymus derived lymphocyte; Thymus-Dependent Lymphocytes; Tissues; trafficking; transfer of a gene; Transgenic Mice; United States; university student; urogenital system (genital part); Vaccines; Vagina; Vaginal; Veiled Cells; Venereal Diseases; Venereal Disorders; venereal herpes; Venereal Infections; Viral; Viral Burden; Viral Diseases; Viral Drug Resistance; viral infection; Viral Load; Viral Load result; Viral Shedding; Viral Sheddings; Virus; Virus Diseases; virus infection; Virus Shedding; Virus Sheddings; Virus-Genital Herpes; Viruses, General; Wild Type Mouse; Woman; youngster
Project start date: 2007-07-01
Project end date: 2011-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2011
5R01AI067309-03 (2009): $286384
Sponsored Links Excellgen http://Excellgen.com
REGULATION OF VENTRAL HIPPOCAMPAL SYSTEM FUNCTION
J Daniel, Assistant Professor
University Of Texas Hlth Sci Ctr San Antcity: San Antonio country: United States (us)
Grant 5R01MH090067-02 from National Institute Of Mental Health
Abstract: Schizophrenia is a devastating psychiatric condition that affects approximately 0.4-1% of the US population. Substantial convergent evidence from drug studies, therapeutic treatments, and human imaging studies demonstrate that psychosis in schizophrenia is associated with a dysregulation of subcortical dopamine system function. This dopamine hypothesis of schizophrenia has one major caveat in that there appears to be no specific pathology in the midbrain dopamine system itself. Thus it is likely that it is the afferent regulation of the dopamine system that is dysfunctional in schizophrenia. Two such inputs are the medial prefrontal cortex (thought to be largely associated with cognitive deficits) and the hippocampus, a temporal lobe structure principally associated with learning and memory. Alterations in hippocampal structure and function in schizophrenia are consistently demonstrated in postmortem and neuro-imaging studies. Furthermore, there is increasing evidence for baseline hippocampal hyperactivity in human schizophrenia patients that is correlated with levels of psychosis. Consistent with this, we have recently reported baseline hyperactivity in the ventral hippocampus in a developmental disruption rodent model, namely MAM G17, that appears to be the driving force behind the dopamine hyperfunction in this model. Thus, attenuation of ventral hippocampal output may act to normalize dopamine transmission and may provide a novel therapeutic target. We plan to examine this model utilizing distinct approaches aimed specifically at modulating ventral hippocampal activity along the following Specific Aims 1) Examine potential novel methods for regulating vHipp function in MAM and saline rats, 2) Determine how these methods for attenuating vHipp activity alter dopamine system function in MAM and saline rats, and 3) Determine whether attenuating vHipp system function can reverse behavioral deficits associated with schizophrenia in the MAM model. Examining the functional interactions among these systems and how disruption within this circuit affects information processing, neurochemistry and behavior is central to our ultimate goal of identifying new and improved therapeutic agents
Keywords: Ablation; Affect; Agonist; Animal Model; Attention; Attenuated; attenuation; Autopsy; base; Behavior; Behavioral; brain pathway; Brain region; Chemicals; Cognitive; Cognitive deficits; Complex; Data; Deep Brain Stimulation; Development; Disease; Dopamine; dopamine system; driving force; Frequencies (time pattern); Functional disorder; gamma-Aminobutyric Acid; Glutamates; Goals; Hippocampus (Brain); Human; human data; Hyperactive behavior; Image; improved; information processing; Interneurons; Learning; Medial; Mediating; Memory; memory recall; Mental disorders; mesolimbic system; Methods; Midbrain structure; Modeling; Neurobiology; neurochemistry; neuron development; Neurons; neurophysiology; neuropsychiatry; neurotransmission; new therapeutic target; novel; Nucleus Accumbens; Operative Surgical Procedures; Output; Parvalbumins; Pathology; Patients; Pharmaceutical Preparations; Pharmacological Treatment; Population; Prefrontal Cortex; Principal Investigator; programs; Property; psychostimulant; Psychotic Disorders; Rattus; receptor; Recruitment Activity; Regulation; Reporting; response; Rest; Rodent Model; Role; Saline; Schizophrenia; Secondary to; Signal Transduction; Structure; Symptoms; System; Temporal Lobe; Temporal Lobe Epilepsy; Therapeutic; Therapeutic Agents; Therapeutic Studies; Toxic effect; transmission process; Work
Relevance: Schizophrenia is a devastating psychiatric condition that affects 0.4% of the US population. One brain region commonly associated with this disease is the hippocampus. In this project we will examine novel methods for altering activity in this brain region in an attempt to better understand disease pathophysiology, with the ultimate goal being the development of improved therapeutic agents
Project start date: 2010-07-01
Project end date: 2015-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-07-070
5R01MH090067-02 (2011): $330784
MULLER GLIA AND RETINA REGENERATION
J Daniel, Professor/research Professor
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Grant 5R01EY018132-04 from National Eye Institute
Abstract: Retinal degenerations are the most common cause of blindness in the Western world. Over 5% of the population will, at some point in their lives, suffer from retinal degeneration. Although we have made great strides in identifying genes responsible for much retinal degeneration, strategies for repairing the diseased retina have remained elusive. Recent studies suggest that the mammalian retina harbors progenitors that can restore lost neurons, albeit with very low efficiency. One goal of neurobiologists studying the retina is to identify strategies for inducing a regenerative response that would facilitate repair of the diseased or damaged retina. Model systems, like zebrafish, offer an exceptional advantage when studying retinal regeneration, in part because of their robust regenerative powers. The mechanism by which fish successfully regenerate a damaged retina is not known, although we recently provided evidence suggesting Muller glia are involved. The long term goal of our research is to ascertain the role Muller glia play in zebrafish retinal regeneration and identify the mechanisms by which these cells contribute to retinal repair. Ultimately it is hoped that this information can be applied to the mammalian retina to improve its regenerative ability. We propose to use a combination of cell and molecular biological approaches to investigate the role Muller glia play in retinal regeneration. Specifically we propose to 1) Determine if Muller glia mediate regeneration of all major retinal cell types in the regenerating retina; 2) Determine if Muller glia are necessary for successful retina regeneration; 3) Identify the genetic changes that impart a progenitor status to Muller glia following retinal injury; and 4) Identify mechanisms underlying a1T gene expression and Muller glia dedifferentiation during retinal regeneration. These studies will form the foundation for future investigations of Muller glia as a source of cells for retinal repair in fish and mammals. Unlike mammals, injury to the fish retina initiates a regenerative response that can restore lost visual function. Our research aims to identify the mechanisms underlying successful retina regeneration in fish. We propose that these mechanisms may suggest new strategies for improving repair of the damaged or diseased mammalian retina
Keywords: Active Follow-up; Applications Grants; Biological; Biological Models; Blindness; Brachydanio rerio; cell type; Cells; Danio rerio; DNA Recombination; DNA recombination (naturally occurring); experiment; experimental research; experimental study; Fishes; follow-up; Foundations; Future; Gene Expression; Genes; Genetic Alteration; Genetic Change; Genetic defect; Genetic Recombination; Genome; genome mutation; Glia; Glial Cells; Goals; Grant Proposals; Grants, Applications; heavy metal lead; heavy metal Pb; improved; injured; Injury; Investigation; Kolliker`s reticulum; Label; Lead; Life; Mammalia; Mammals; Mammals, General; Maps; Mediating; Model System; Models, Biologic; Molecular; multipotent progenitor; Multipotent Stem Cells; Mutation; Natural regeneration; Nerve Cells; nerve cement; Nerve Unit; Neural Cell; Neurocyte; Neuroglia; Neuroglial Cells; neuronal; Neurons; Non-neuronal cell; Pb element; Play; Population; Process; progenitor; Proliferating; Recombination; Recombination, Genetic; regenerate; Regeneration; regenerative; repair; repaired; Research; research study; response; Retina; retina degeneration; Retinal; Retinal Degeneration; retinal degenerative; retinal neuron; retinal regeneration; Role; Sight; Signaling Molecule; social role; Source; stem cell population; System; System, LOINC Axis 4; Testing; transcription factor; Tubulin; Vision; Western World; Zebra Danio; Zebra Fish; Zebrafish
Project start date: 2007-04-01
Project end date: 2012-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2012
5R01EY018132-04 (2010): $367850
CENTRAL NERVOUS SYSTEM MECHANISMS IN KNEE OSTEOARTHRITIS (KOA)
J Daniel, Professor
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Grant 1R01AR060392-01A1 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Recent studies suggest the lifetime risk for symptomatic knee OA (KOA) is 45%1. An aging population and increasing rates of obesity contribute to dramatic increases in the prevalence of this condition. Historically, the "disease" of OA is viewed primarily as damage to the cartilage and bone. As such, the magnitude of damage or inflammation of these structures should predict symptoms. Population-based studies suggest otherwise; 30- 50% of individuals with moderate to severe radiographic changes of OA are asymptomatic, and approximately 10% with moderate to severe knee pain have normal radiographs2,3. Psychological factors do account for some 4,5 of this variance in pain and other symptoms, but only to a small degree . This failure of peripheral damage, inflammation, or even psychological factors to explain the presence, absence, or severity of chronic pain should not be surprising. To date, no chronic pain state involves a strong relationship between peripheral factors and the level of pain reported. We hypothesize that, although peripheral nociceptive input and to a lesser extent psychological factors are important in leading to pain and symptom expression in KOA, some patients possess varying degrees of non-psychological central nervous system (CNS) factors which play an equally or even more prominent role in the expression of pain and co-morbid symptoms 6-8. Broadly within chronic pain states, subsets of patients have been identified that have prominent CNS mechanisms (as opposed or in addition to, peripheral damage) playing important roles in pain perception. Such factors include diffuse hyperalgesia or allodynia, and/or a lack of endogenous descending analgesic activity 6,8, 9 . The exploration of these CNS factors has been somewhat limited to date in OA, but evidence is emerging that supports the hypothesis that subsets of OA patients do indeed have these mechanisms operative 10-12. Our hypothesis is further strengthened by studies that have identified co-morbid somatic symptoms known to be associated with central pain conditions (e.g., fatigue, sleep problems) to be commonly present in OA 13, 14. Finally, recent RCTs have demonstrated that compounds that alter pain neurotransmitters centrally such as 15,16 serotonin and norepinephrine (e.g., duloxetine, tricyclics) are efficacious in OA . We will identify the role that CNS factors are playing in KOA by first showing that subsets of patients have symptom patterns and experimental sensory testing abnormalities consistent with having a "central" component to their pain. We will then demonstrate the utility of these measures by showing that individuals with KOA with central pain will respond poorly to knee arthroplasty. Such a study possesses 17 high public health impact given the high "failure" rate of knee arthroplasty . If the status quo in practice for KOA is maintained, demand for knee arthroplasty will increase by an expected 700% in the next 20 years 18. Thus, in addition to this study providing a potential paradigm shift in our thinking regarding the "disease" of OA, this study also develops practical clinical tools for identifying the presence of centrally-enhanced pain in OA. This study aims to examine why individuals with osteroarthritis of the knee respond poorly to knee arthroplasty. With demand for this procedure increasing, and an aging population, it is important to examine why there is such a high failure rate in this population
Keywords: Accounting; aging population; allodynia; Analgesics; Arthroplasty, Replacement, Knee; base; bone; Cartilage; central pain; chronic pain; Clinical; Descriptor; Diffuse; Disease; duloxetine; Failure (biologic function); Fatigue; Genetic; Hyperalgesia; Individual; Inflammation; Knee; Knee Osteoarthritis; knee pain; Lifetime Risk; Measures; Neuraxis; Neuropathy; Neurotransmitters; Nociception; Norepinephrine; Obesity; Operative Surgical Procedures; Pain; Patients; Pattern; Perception; Peripheral; Persistent pain; Play; Population; population based; Prevalence; Procedures; Prospective Studies; Psychological Factors; public health medicine (field); Reporting; Role; Sensory; Serotonin; Severities; Sleep Disorders; Structure; Symptoms; Testing; Thinking, function; tool
Relevance: This study aims to examine why individuals with osteroarthritis of the knee respond poorly to knee arthroplasty. With demand for this procedure increasing, and an aging population, it is important to examine why there is such a high failure rate in this population
Project start date: 2011-08-01
Project end date: 2016-05-31
Budget start date: 1-AUG-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-10-006
1R01AR060392-01A1 (2011): $679900
BIOCHEMISTRY OF GROWTH REGULATION AND ONCOGENESIS
J Daniel, Professor Of Chemistry/biochemistry
University Of California San Diegocity: La Jolla country: United States (us)
Grant 3T32CA009523-27S1 from National Cancer Institute
Abstract: PURPOSE AND PROGRAM CHARACTERISTICS This proposal represents a continuation of a training program in the biochemistry of growth regulation and oncogenesis, now entering its twenty-fourth year. The training faculty is drawn from the UCSD Department of Chemistry and Biochemistry, from the Division of Biological Sciences, and from the School of Medicine. All members of the training faculty are also members of the Moores UCSD Cancer Center and participate in their programs. In this renewal, the training faculty has been strengthened by the addition of 14 outstanding faculty members, bringing the total number of faculty members to 32, including six members of the National Academy of Sciences. The training faculty is organized into four research areas focusing on kinases, phosphatases, and their signaling pathways; the control of cell cycle progression, DNA checkpoints, and apoptosis; transcription factors and their signaling pathways; and the chemistry of cancer therapeutics. This program exhibits a unique emphasis on structural and molecular approaches. Training involves formal courses, journal clubs, annual symposia, and a variety of events to promote program cohesion and interaction. TRAINEES Our current and past trainees have excellent records of research accomplishments. We have requested seven postdoctoral positions in Years 24 and 25, and increasing thereafter to reach ten postdoctoral trainees in Year 28. No change is requested in predoctoral slots. The requested slots are justified by the enormous expansion currently underway at UCSD in undergraduate and graduate enrollments, by the ability of the training faculty to recruit outstanding predoctoral and postdoctoral trainees to their labs, and by the highly interactive nature of the training labs that collectively provide a superb training environment. Predoctoral trainees will be drawn from graduate students accepted into the Department of Chemistry and Biochemistry and, with this renewal, from graduate students in Biological Sciences. Postdoctoral trainees will be selected from postdoctoral candidates applying for positions in the laboratories of the training faculty. All trainees accepted into our program are expected to have strong backgrounds in chemistry, biochemistry, and molecular and cell biology. PROGRAM CHANGES This renewal highlights several changes to the program, including greater program integration with the new Rebecca and John Moores UCSD Cancer Center; increased cancer relevance of our faculty; doubling of our predoctoral pool; proposed change to increase the proportion of postdoctoral trainees; definition of research focus groups; endorsements from key administrative leaders; and improved mechanisms to build program cohesion
Keywords: Admission; Admission activity; Anabolism; Animal Virology; Animal Viruses; anticancer research; Apoptosis; Apoptosis Pathway; Appointment; Area; Articulation; base; Basic Cancer Research; Beer; Bio-Informatics; Biochemical; Biochemistry; bioengineering; bioengineering/biomedical engineering; Bioinformatics; Biologic Sciences; Biological; Biological Factors; Biological Models; Biological Sciences; biological signal transduction; biological systems; Biology; Biomedical Engineering; biosynthesis; Cancer Biology; cancer cell; Cancer Center; Cancer Center Director; Cancer Genes; Cancer Induction; cancer research; cancer research center director; Cancer-Promoting Gene; Cancers; carcinogenesis; Catalysis; cell biology; Cell Communication and Signaling; Cell Cycle Control; Cell Cycle Progression; Cell Cycle Regulation; Cell Cycle Regulation, Including Apoptosis; Cell Death, Programmed; Cell division; Cell Function; Cell Locomotion; Cell Migration; cell motility; Cell Movement; Cell physiology; Cell Process; Cell Signaling; Cellular biology; Cellular Function; Cellular Matrix; Cellular Migration; Cellular Physiology; Cellular Process; Cellular Transformation; Chair; Chairman; Chairperson; Chairwoman; Characteristics; Chemistry; Chemistry, Biological; chemotherapy; Chromatin Structure; Cloning; coenzyme analog; Coenzymes; Cofactors, Enzyme; cohesion; Commit; Committee Members; Communities; computational modeling; computational models; computational simulation; computer based models; Computer Graphics; Computer Simulation; computerized modeling; Computerized Models; computerized simulation; conference; Consultations; Curriculum; Cytoskeletal System; Cytoskeleton; Data; Deoxyribonucleic Acid; Development; Director of Cancer Research Center; DNA; DNA biosynthesis; DNA Molecular Biology; DNA Replication; DNA Synthesis; Doctor of Philosophy; doubt; E-Mail; EC 2.7; Educational Curriculum; Educational workshop; Electronic Mail; Email; enroll; Enrollment; Ensure; Environment; Enzymes; Event; Exhibits; expectation; experience; Exposure to; Factor, Biologic; Faculty; Focus Groups; Food; Foundations; Funding; Future; Gene Action Regulation; Gene Expression; Gene Expression Regulation; Gene Organization; gene product; Gene Products, RNA; Gene Regulation; Gene Regulation Process; Gene Structure; Gene Structure/Organization; Gene Transcription; Generalized Growth; Genetic; Genetic Structures; Genetic Transcription; Genomics; Goals; graduate student; Grant; Group Meetings; Growth; Hearing; hearing perception; Histopathology; Hour; human disease; Human Genetics; Immunology; Immunology (Including BRMP); Immunology (NCI Program); improved; in silico; Individual; inhibitor; inhibitor/antagonist; Institutes; Institution; interest; Intracellular Communication and Signaling; intracellular skeleton; Investigators; Joints; journal article; Journal Article; Journal Article (PT); Journal Article [Publication Type]; Journals; Judgment; Kinases; Knowledge; Laboratories; Lectures; lectures; Lectures (PT); Lectures [Publication Type]; Life Sciences; Lobbying; Macromolecular Protein Complexes; macromolecule; Magazine; malignancy; Malignant Cell; Malignant Neoplasms; Malignant Tumor; Mammalia; Mammals; Mammals, General; Mathematical Model Simulation; Mathematical Models and Simulations; Medical; medical schools; Medicine; meetings; member; Mentors; metaplastic cell transformation; Methods; Methods and Techniques; Methods, Other; Model System; Models, Biologic; Models, Computer; Molecular; Molecular Analysis; Molecular Biology; Molecular Genetic; Molecular Genetics; Molecular Motors; molecular pathology; molecular recognition; Motility; Motility, Cellular; Multiprotein Complexes; Names; National Academy of Sciences; National Academy of Sciences (U.S.); Natural Products; Natural Products Chemistry; Natural Sciences; Nature; neoplasm/cancer; Nucleic Acid Folding; Nucleic Acids; Oncogenes; Oncogenesis; Oncornaviruses; Oncovirinae; Oncoviruses; ontogeny; Oral; Oral Examination; Paper; pathway; Pathway interactions; Peptide Biosynthesis, Ribosomal; Peptide Domain; Peptide/Protein Chemistry; Ph.D.; PhD; Phosphatases; Phosphohydrolases; Phosphomonoesterases; Phosphoric Monoester Hydrolases; Phosphotransferases; Position; Positioning Attribute; post-doc; post-doctoral; post-doctoral training; Postdoc; Postdoctoral Fellow; postdoctoral training; posters; Posters; Posters [Publication Type]; pre-doc; pre-doctoral; predoc; predoctoral; Procedures; programs; Programs (PT); Programs [Publication Type]; Progress Reports; prospective; Protein Biochemistry; Protein Biosynthesis; Protein Biosynthesis, Ribosomal; Protein Chemistry; Protein Domains; protein protein interaction; protein structure; protein synthesis; Protein Synthesis, Ribosomal; Protein/Amino Acid Biochemistry; Proteins; Qualifying; R01 Mechanism; R01 Program; Reaction; Reading; Rebecca and John Moores UCSD Cancer Center; Rebecca and John Moores University of California at San Diego Cancer Center; receptor mediated endocytosis; Records; recruit; Recruitment Activity; Regulation; Reporting; Research; Research Associate; Research Grants; Research Institute; Research Personnel; Research Project Grants; Research Projects; Research Projects, R-Series; Researchers; response; Ribonucleic Acid; RNA; RNA Expression; RNA Tumor Viruses; RNA, Non-Polyadenylated; Rotation; RPG; Running; SCHED; Schedule; Science; Science of Chemistry; Science of Medicine; Science of Virology; screening; Screening procedure; screenings; Series; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signal Transduction Systems; Signaling; Simulation, Computer based; skills; small molecule; Solid; sound perception; Specialist; Structure; Students; Subcellular Process; Survey Instrument; Surveys; symposium; Techniques; Tertiary Protein Structure; Testing; The UCSD Cancer Center; Therapeutic; Time; Tissue Growth; Training; Training Programs; Training Support; Transcription; transcription factor; Transcription, Genetic; Transforming Genes; Translations; Transphosphorylases; Travel; tumorigenesis; Uncertainty; United States National Academy of Sciences; Viral; Virology; virology; virtual simulation; Virus; Viruses, General; Visit; web site; Work; Workshop
Project start date: 1993-03-01
Project end date: 2012-12-31
Budget start date: 1-JAN-2011
Budget end date: 31-DEC-2011
PFA/PA: PA-06-468
3T32CA009523-27S1 (2011): $59195
5T32CA009523-27 (2011): $574184
3T32CA009523-27S2 (2011): $58365
3T32CA009523-27S3 (2011): $37207
CLINICAL TRANSPLANTATION EXPLOITING NK CELL ACTIVITY
J Daniel
University Of Minnesota Twin Citiescity: Minneapolis country: United States (us)
Abstract: PROJECT SUMMARY (See instructions) In the previous funding period we identified immunogenetic loci that an unrelated donor (URD) KIR B haplotype yields statistically significant and meaningful improvement in protection against relapse and relapse free-survival fpr patients with AML. New analyses have refined the favprable KIR B loci to those which encode donor KIR activating receptors associated with improved clinical outcome. We will further explore the clinical importance of these genetic loci by prospectively evaluating the logistics and clinical impact of donor selection by KIR genotyping for URD hematopoietic cell transplantation (HCT) in AML. We hypothesize that amongst URD with optimal HLA matching, donor KIR genotyping can identify better donors likely to yield improved clinical outcome. In conjunction Project 1, we will explore the functional significance of these KIR B loci, their interaction with HLA Class I and allelic polymorphism in the KIR regions to further refine our understanding and methods for prospective donor selection. With Project 2 we will analyze NK cell functional development over time after URD HCT and correlation of this NK development with the complications following HCT including engraftment, GVHD, infections, relapse and survival. These unique analyses accompany a BMT CTN prospective trial testing blood vs. marrow as a graft source for URD HCT and offer a unique platform for understanding how NK cell development, KIR genotyping and functional immune reconstitution can modify post-transplant complications and outcomes. Additionally, the fundamentals of NK cell therapy to reduce recurrence of resistant leukemia and improve sun/ival after transplantation will.be directly tested. In a multicenter trial of reduced intensity haploidentical HCT plus donor NK infusions for resistant AML we will evaluate the direct impact of NK cell therapy on highly resistant leukemia. Prospective clinical trials will define elements essential for safer application of this treatment. Overall, these studies can improve the survival and relapse protection following allogeneic HCT for AML. Our studies outline new and exciting opportunities to improve donor selection and maximize the safety and effectiveness of allotransplantation
Keywords: Acute leukemia; Adoptive Transfer; Alleles; Allogenic; Allografting; Antithymoglobulin; base; Blood; Blood Tests; Cell physiology; Cell Therapy; Cell Transplantation; Chemotherapy-Oncologic Procedure; Chromosomes, Human, Pair 19; Clinical; Clinical Data; Clinical Trials; Clinical Trials Network; cohort; Cohort Studies; Communities; Complex; Data; Development; direct application; Disease remission; Disease-Free Survival; Donor person; Donor Selection; Effectiveness; Elements; Engraftment; Enrollment; Ensure; Event; fighting; Funding; Genes; Genetic; genetic element; Genetic Polymorphism; Genotype; Goals; Graft-vs-Host Disease; Haplotypes; Hematopoietic; high risk; Immune; Immunogenetics; immunoglobulin receptor; improved; in vivo; Incidence; Individual; Infection; Infusion procedures; Instruction; Interleukin-2; International; Investigation; Knowledge; Lead; leukemia; Location; Logistics; Marrow; Methods; Morbidity - disease rate; Mortality Vital Statistics; Multicenter Trials; Natural Killer Cells; novel; Outcome; Patients; Pattern; peripheral blood; phase 2 study; Phase II Clinical Trials; Population; programs; prospective; Prospective Studies; Protocols documentation; Randomized; receptor; Receptor Gene; reconstitution; Recovery; Recurrence; Recurrent disease; Refractory; Regimen; Regulation; Relapse; Research; Resistance; Risk; Role; Safety; Sampling; Series; Site; Source; success; Testing; The Sun; Time; Translating; Transplantation; Whole-Body Irradiation; Work
Relevance: Patients with advanced acute leukemia can be treated with donor transplantation, but improved means to select donors, enhance immune recovery after transplantation and control the risks of relapse have direct application in improving patient survival. These studies will change transplant practice to utilize NK cells to make transplantation safer and more effective
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
5P01CA111412-07_5847 (2011): $421429
MECHANOBIOLOGY OF LUNG FIBROSIS
J Daniel
Harvard University (sch Of Public Hlth)city: Boston country: United States (us)
Grant 5R01HL092961-03 from National Heart, Lung, And Blood Institute
Abstract: Progressive fibrosis is a hallmark of interstitial lung diseases such as idiopathic pulmonary fibrosis. Fibrosis stiffens the lung parenchyma. Preliminary data provided in this proposal demonstrate that normal lung parenchymal tissue is more compliant than previously predicted, and that highly localized increases in the stiffness of fibrotic lesions are much greater than previously recognized. Fibroblasts grown on substrates as stiff as fibrotic lesions engage in rapid proliferation and abundant matrix synthesis; in marked contrast these behaviors are largely suppressed on substrates as compliant as normal lung tissue. The central hypothesis that we pose based on these data is that the mechanical environment present in lung fibrosis triggers a "fibrogenesis program" in resident fibroblasts that promotes feedback amplification of the disease. We propose four specific aims (1) quantify the stiffness of the parenchyma in normal lung tissue and developing and established fibrotic lesions; (2) test whether the transition in matrix stiffness from normal to fibrotic levels is a necessary precondition for lung fibroblast proliferation and fibrogenic activation; (3) test the role played by cytoskeletal dynamics and serum response factor activation in driving stiffness-dependent fibroblast biology; and (4) identify key transcription factors coordinating fibroblast transitions between quiescent and fibrogenic states when transferred between compliant and stiff matrices. These aims will be carried out in novel 2D and 3D models that allow clear delineation of the effects of lung matrix stiffness on key fibrogenic behaviors of lung fibroblasts. Throughout the experimental plan we will examine the interplay between matrix stiffness and the soluble environment present in fibrosis as it impacts on fibroblast biology. This research will generate novel insights into the molecular mechanisms of stiffness-dependent fibroblast activation in fibrotic lungs, and identify critical regulators of fibrogenesis suitable for therapeutic targeting. Lung fibrosis stiffens affected tissue. The experiments proposed here will test whether pathophysiological changes in stiffness promote fibrosis by stimulating lung fibroblasts. Understanding the role mechanical factors play in fibroblast activation could lead to new strategies to treat fibrosis
Keywords: Actins; Affect; Apoptosis; Atomic Force Microscopy; Automobile Driving; base; Behavior; Bioinformatics; Biological; Biology; Bleomycin; Bronchoalveolar Lavage Fluid; Cell Culture Techniques; Cells; cytokine; Data; Deposition; Disease; Employee Strikes; Environment; Exhibits; F-Actin; Feedback; Fibroblasts; fibrogenesis; Fibrosis; Focal Adhesions; G Actin; Gases; Gel; Gene Expression; Growth; Hamman-Rich syndrome; Health; Human; Immunofluorescence Immunologic; insight; Interstitial Lung Diseases; Knowledge; Lead; Lesion; Link; Lung; lung injury; Measurement; Mechanics; Modeling; Molecular; Mus; Myofibroblast; novel; Outcome; Phenotype; Play; polyacrylamide gels; polymerization; preconditioning; programs; protein expression; public health relevance; Pulmonary Fibrosis; Regulator Genes; Research; research study; response; RNA Interference; Role; Saline; Serum; Serum Response Factor; Signal Transduction; Stress Fibers; Structure of parenchyma of lung; Testing; therapeutic target; three-dimensional modeling; Tissues; transcription factor
Relevance: Lung fibrosis stiffens affected tissue. The experiments proposed here will test whether pathophysiological changes in stiffness promote fibrosis by stimulating lung fibroblasts. Understanding the role mechanical factors play in fibroblast activation could lead to new strategies to treat fibrosis
Project start date: 2009-08-06
Project end date: 2013-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01HL092961-03 (2011): $413123
NEURONAL INTEGRATION IN THE NEOCORTEX
J Daniel, Professor
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5R01NS019950-27 from National Institute Of Neurological Disorders And Stroke
Abstract: Perception and other cognitive functions such as planning, thought and learning reflect processing of complex information by the cerebral neocortex. Adaptive function depends critically on inputs from subcortical centers and on subsequent signal processing by appropriately organized cortical microcircuits. The proposed work asks how information is faithfully transferred from one part of the brain to the next and how signal-transforming feedforward networks become refined by developmental experience. Using the rodent whisker-to-barrel system, we will examine thalamocortical microcircuits in normally- reared animals and in animals raised with abnormal tactile experience produced by simple trimming of whisker hairs during early postnatal life. Using quantitative sensory physiology methods and single-cell recordings, including anatomical labeling of functionally characterized thalamocortical neurons, we will evaluate hypotheses derived from a long-standing model of thalamocortical function in somatosensory and visual systems. The model postulates that response tuning of cortical layer 4 neurons reflects specific thalamic inputs and that local circuitry dynamically adjusts receptive field properties by regulating overall network excitability. Specific experiments will investigate the innervation pattern of thalamocortical axons, their functional impact, and their experience-dependent developmental plasticity. Results are expected to establish key mechanisms of thalamocortical function and development shared by different systems in different species. Normal brain function depends critically on faithful transmission of information from one part of the brain to the next. Early postnatal experience influences the development of circuits that receive and process the signals. This research plan investigates sensory signaling and experience-dependent plasticity in order to understand how abnormally functioning cortical microcircuits might contribute to impairments in perceptual/motor and other cognitive behaviors
Keywords: Address; Adult; Animals; Axon; base; Behavior; Brain; Brain Part; Cells; Cerebral cortex; Cerebrum; Cognitive; cognitive function; Complex; computerized data processing; Cortical Column; deprivation; Development; developmental plasticity; excitatory neuron; experience; Exploratory Behavior; Growth; Hair; Impairment; Individual; inhibitory neuron; juvenile animal; Label; Learning; Life; Location; Measures; Mediating; Methods; Modeling; Morphology; Motor; Neocortex; Neonatal; nerve supply; Neurons; Pattern; Perception; postnatal; preference; Process; Property; public health medicine (field); public health relevance; receptive field; Research; research study; response; Rodent; Sensory; Sensory Physiology; Signal Transduction; somatosensory; stem; Stimulus; Structure; Synapses; synaptic depression; System; Tactile; Thalamic structure; transmission process; Vibrissae; Visual system structure; Work
Relevance: RELEVANCE TO PUBLIC HEALTH Normal brain function depends critically on faithful transmission of information from one part of the brain to the next. Early postnatal experience influences the development of circuits that receive and process the signals. This research plan investigates sensory signaling and experience-dependent plasticity in order to understand how abnormally functioning cortical microcircuits might contribute to impairments in perceptual/motor and other cognitive behaviors
Project start date: 1983-07-01
Project end date: 2014-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-07-070
5R01NS019950-27 (2011): $298266
Sponsored Links Excellgen http://Excellgen.com
BIOPREP: BIOLOGY PARTNERSHIP IN RESEARCH AND EDUCATION PROGRAM
J Daniel, Lecturer
State University New York Stony Brookcity: Stony Brook country: United States (us)
Grant 5R25GM050070-12 from National Institute Of General Medical Sciences
Abstract: Our commitment is to increase the number of students from two-year institutions belonging to underrepresented or health disparities populations who obtain undergraduate degrees in the sciences and pursue graduate education and careers in the biomedical sciences. This will be accomplished by continuing to develop the partnership between Stony Brook University and Nassau, Suffolk, and Queensborough Community Colleges. The goals of our program are 1) to identify eligible students at the two-year institutions interested in the biomedical sciences; 2) to motivate these students to seek careers in the biomedical sciences; 3) to help them gain confidence in their abilities by exposure to research scientists and laboratory techniques; 4) to create a supportive environment, through mentoring, counseling and group activities; 5) to promote an awareness of the opportunities available in the biomedical sciences; and 6) to create a model for partnerships between two-year institutions and research universities that will outline how other institutions may meet these objectives. Our objectives are being met by recruiting up to twenty-four eligible students into the program each year and 1) providing these students with the techniques and skills to understand and use the basic tools of molecular biology; 2) providing students with research projects; 3) providing students with the opportunity for internships with research faculty; 4) providing students with up-to-date knowledge from current researchers; 5) providing students with tutoring from more advanced undergraduate and graduate students; 6) providing mentors for all accepted students; 7) providing students with opportunities to present their work at local and national meetings; 8) educating the students about and easing the transition process from a two-year to a four-year institution; 9) providing financial support for BioPREP students who transfer to Stony Brook; and 10) assisting students in applying to graduate school or in obtaining employment. This project encourages outstanding underrepresented two-year college students to pursue careers in biomedical research. This will increase the numbers of underrepresented students in the research pipeline. This supports the national goal of bringing a more diverse population into biomedical research careers
Keywords: Awareness; Bachelor`s Degree; base; Biology; Biomedical Research; career; college; Communities; County; Educational aspects; Educational Background; Employment; Environment; Exposure to; Face; Faculty; Financial Support; Goals; Graduate Education; graduate student; group counseling; health disparity; Health Personnel; Healthcare Systems; innovation; Institution; interest; Internships; Journals; Knowledge; Laboratories; Laboratory Scientists; Learning; Medicine; meetings; Mentors; Modeling; Molecular Biology; New York; Population; Population Heterogeneity; Process; programs; Recruitment Activity; Research; Research Personnel; Research Project Grants; Schools; Science; skills; Students; Techniques; tool; United States; Universities; university student; Work
Project start date: 1993-09-01
Project end date: 2012-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PAR-07-039
5R25GM050070-12 (2011): $228018
DESCENDING NORADRENERGIC AND SEROTONERGIC ANALGESIC ACTIVITY IN IC/PBS
J Daniel, Professor
University Of Michigan At Ann Arborcity: Ann Arbor country: United States (us)
Abstract: Our central hypothesis in the Michigan MAPP Discovery Site is that a subset of women with IC/PBS has a "central" problem in pain or sensory processing, as occurs in fibromyalgia (FM), rather than a disorder confined to the bladder. Project 3 will further explore the precise reason(s) for the augmented pain and sensory processing seen in IC/PBS. In this study, a specific molecular probe will be administered to IC/PBS patients to test whether a subset of these individuals has attenuated descending analgesic activity that is restored when they are administered a selective noradrenergic/serotonergic reuptake inhibitor (NSRI), milnacipran. We hypothesize that a subset of IC/PBS patients has attenuated noradrenergic and serotonergic activity in descending analgesic pain processing pathways, and that this will be identifiable both by experimental pain testing and fMRI, and that this will be reversed when these individuals are administered a specific norepinephrine-serotonin reuptake inhibitor, milnacipran. The specific aims of this study are to 1) To demonstrate that some 1C patients have attenuated descending noxious inhibitory control (DNIC), as has been identified in FM, 2) To show that when 1C patients are given milnacipran, a norepinephrine-serotonin reuptake inhibitor, these individuals will have restoration of DNIC, 3) To demonstrate that the improvements of DNIC identified by experimental pain testing can also be shown using fMRI, 4) To show that the improvements in DNIC with the administration of milnacipran to IC/PBS patients are accompanied by improvements in clinical symptoms, and 5) To show that genetic polymorphisms involving catachol-O-methyl-transferace (COMT) and beta 2 and -3 adrenoreceptors. This study will not only probe a specific mechanism in 1C that has been shown to be dysfunctional in FM and IBS (the absence of appropriate descending analgesic activity), but also demonstrate "proof-of-concept" that this abnormality is due to a specific molecular mechanism. Although dual reuptake inhibitors, e.g., tricyclics and SNRI/ NSRIs are agreed to be efficacious in central pain states and this activity has been suspected to be due to augmented descending analgesic activity, this mechanism has never been definitively supported, thus these studies will be important in the broader pain field as well as advancing knowledge in 1C
Keywords: Adverse drug effect; Adverse effects; Affect; Amitriptyline; Analgesics; Attenuated; Binding (Molecular Function); Biological; biomarker; Bladder; central pain; cholinergic; Cholinergic Receptors; chronic pain; Clinical; clinical effect; clinical practice; Clinical Trials; Cognitive; cohort; Complex; Data; design; Disease; drug efficacy; Drug usage; Fatigue; Fibromyalgia; Functional Magnetic Resonance Imaging; Future; Genetic; Genetic Polymorphism; Hypertension; improved; Individual; inhibitor/antagonist; Knowledge; Measures; Medicine; Methods; Michigan; midalcipran; Molecular; Molecular Probes; Multicenter Studies; Nature; neuroimaging; noradrenergic; Norepinephrine; Outcome; Outcome Measure; Pain; Pain Threshold; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; pressure; Process; Process Measure; Property; psychologic; Research; Research Methodology; restoration; reuptake; Role; Selective Serotonin Reuptake Inhibitor; Sensory Process; Serotonin; Site; Sodium Channel; Symptoms; Techniques; Testing; Toxic effect; Universities; Woman; Work
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5U01DK082345-04_0003 (2011): $423921
CORNEAL LYMPHATICS & ADAPTIVE IMMUNITY
J Daniel
University Of Oklahoma Hlth Sciences Ctrcity: Oklahoma City country: United States (us)
Grant 1R01EY021238-01 from National Eye Institute
Abstract: Neovascularization in the normally avascular cornea can lead to a compromised visual axis. Within the term "neovascularization" is a blood component referred to as hemangiogenesis and a lymphatic component referred to as lymphangiogenesis. Relative to herpes simplex virus type 1 (HSV-1) infection, only hemangiogenesis has been investigated. Recently, we have initiated a study on lymphangiogenesis following corneal infection with HSV-1 and discovered this process precedes hemangiogenesis. More importantly, we have discovered the genesis of lymphatic vessels in the cornea proper in response to HSV-1 infection operates thru a unique pathway distinct from what has been described following corneal transplantation. Specifically, we have found a robust vascular endothelial growth factor (VEGF) A response by corneal epithelium infected with HSV-1 elicits lymphangiogenesis thru a VEGF receptor 2 (VEGFR2)-mediated pathway. This process is independent of VEGF C, VEGF receptor 3, or monocytes/macrophages. We have also found the newly created lymphatic vessels are capable of transporting soluble antigen to the draining lymph node independent of hemangiogenesis. However, what remains unknown is the contribution of the newly created lymphatic conduit to the host immune response within the draining lymph node as well as other inflammatory mediators that contribute to corneal lymphangiogenesis. We hypothesize lymphatic vessel development driven principally by VEGF A and contributing pro-inflammatory molecules generated locally in response to infection are critical for the induction of the adaptive immune response found in the draining lymph node reflected by the severity in the development of herpetic stromal keratitis of HSV-1 infected mice. To address this hypothesis, two specific aims are proposed Specific aim 1 will address the impact of viral replication and pro- inflammatory cytokine expression on corneal lymphangiogenesis following HSV-1 infection. In addition, trafficking of leukocyte subpopulations and antigen will be monitored as well. Specific aim 2 will address the significance of lymphangiogenesis relative to the genesis of the adaptive immune response in the draining lymph node and development of stromal keratitis following HSV-1 infection. It will further address the role of virus-induced VEGF A production on the production of CD4+ and CD8+ T effector cells that contribute in viral surveillance and corneal pathogenesis. It is anticipated in accomplishing these goals, we will eludicate the contribution of pro-lymphangiogenesis factors in the generation of the adaptive immune response critical for the ocular pathology that includes the debilitating and sometimes blinding stromal keratitis. The role of lymphangiogenesis as a central force driving the adaptive immune response to ocular herpes simplex virus type 1 (HSV-1) infection has not been explored. In combination with HSV-1-induced vascular endothelial growth factor (VEGF)-A, a potent immunomodulatory molecule, it is anticipated the study will identify how HSV-1-induced lymphangiogenesis and VEGF-A production influence the immune response to the infection and in so doing, lead to a treatment strategy to alter the host response, attenuate the development of herpetic stromal keratitis, and preserve the visual axis
Keywords: Acute; adaptive immunity; Address; Antibodies; Antigens; ATGN; Attenuated; Blood; Blood monocyte; CD8; CD8B; CD8B1; CD8B1 gene; Cells; chemoattractant cytokine; chemokine; Cornea; corneal; corneal epithelial; corneal epithelium; corneal keratoplasty; corneal transplant; Corneal Transplantation; cytokine; Cytokines, Chemotactic; Development; driving force; Effector Cell; Epithelium, Anterior Corneal; Epithelium, Corneal; Fetal Liver Kinase-1; Flk-1 Protein; Flk-1 Receptor Tyrosine Kinase; Flt-4; FLT4; FLT4 Ligand; FLT4 Protein; FLT4-L; FMS-Related Tyrosine Kinase 4; Generations; Goals; Grafting, Corneal; HBGF; heavy metal lead; heavy metal Pb; herpes simplex i; Herpes Simplex Keratitis; Herpes Simplex Virus 1; Herpes Simplex Virus Type 1; Herpes Simplex, Ocular; herpes virus 1, human; Herpesvirus 1 (alpha), Human; Herpesvirus 1, Human; Herpetic Keratitis; HHV-1; Homologous Chemotactic Cytokines; host response; HSV-1; HSV1; human alphaherpesvirus 1; Human herpes simplex virus type 1; Human herpesvirus 1; Human herpesvirus type 1; Immune; Immune response; immunogen; immunoresponse; Infection; Inflammation Mediators; Inflammatory; Intercrines; intervention therapy; KDR Tyrosine Kinase; Keratitis; Keratitis, Herpetic; Keratoplasty; Kinase Insert Domain Receptor; Lead; Leukocyte Trafficking; lymph gland; Lymph node proper; lymph nodes; Lymphangiogeneses; Lymphangiogenesis; Lymphatic; Lymphatic vessel; LYT3; macrophage; Maintenance; Maintenances; Mammals, Mice; Marrow monocyte; Mediating; Mice; Monitor; monocyte; mouse model; Murine; Mus; neovascularization; ocular herpes; Ocular Pathology; Pathogenesis; Pathology; pathway; Pathway interactions; Pb element; Process; Production; public health relevance; Relative; Relative (related person); response; Reticuloendothelial System, Blood; Reticuloendothelial System, Lymph Node; Role; Severities; SIS cytokines; social role; Structure of corneal epithelium; T cell response; Testing; Therapeutic Intervention; trafficking; Transplantation, Cornea; treatment strategy; Tyrosine-Protein Kinase Receptor FLT4; Vascular Endothelial Cell Growth Factor Receptor; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factor C; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factor Receptor-3; Vascular Endothelial Growth Factor Related Protein; Vascular Permeability Factor Receptor; Vasculotropin; VEGF Receptor Flk-1; VEGF Receptor KDR; VEGF Receptors; VEGF-C; VEGFC; VEGFR; VEGFR-2; VEGFR-3; VEGFR2; VEGFR3; Viral; Virus; Viruses, General; Visual; VPF Receptor; VRP
Relevance: The role of lymphangiogenesis as a central force driving the adaptive immune response to ocular herpes simplex virus type 1 (HSV-1) infection has not been explored. In combination with HSV-1-induced vascular endothelial growth factor (VEGF)-A, a potent immunomodulatory molecule, it is anticipated the study will identify how HSV-1-induced lymphangiogenesis and VEGF-A production influence the immune response to the infection and in so doing, lead to a treatment strategy to alter the host response, attenuate the development of herpetic stromal keratitis, and preserve the visual axis
Project start date: 2010-12-01
Project end date: 2015-11-30
Budget start date: 1-DEC-2010
Budget end date: 30-NOV-2011
PFA/PA: PA-10-067
1R01EY021238-01 (2011): $367939
FERROXIDASE (FET3) AND PERMEASE (FTR1) IN IRON UPTAKE
J Daniel
State University Of New York At Buffalocity: Buffalo country: United States (us)
Grant 5R01DK053820-12 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Multicopper oxidases (MCOs) couple the 4e- reduction of dioxygen to 2H2O with the oxidation of 4 equivalents of a 1-electron donor. A sub-family of these ubiquitous enzymes possesses specificity towards FeII that makes them essential to iron homeostasis in their respective organisms. Our hypothesis is that these ferroxidases function by channeling their FeIII product to a down-stream partner in their iron metabolic pathways. This program has delineated the structure and function in the Fet3p, Ftr1p high-affinity Fe-uptake complex in the Saccharomyces cerevisiae (Sc) plasma membrane (PM). The two fundamental questions addressed in this work are 1) what structural motifs confer on an MCO this specificity for FeII as substrate; and 2) how is the Fet3p ferroxidase reaction coupled kinetically and physically to the membrane permeation of FeIII by Ftr1p. In this application we propose three specific aims. In Aim I we will test our hypothesis of what structural motifs define a ferroxidase, specifically that a cohort of carboxylate side chains maximize three factors that determine e- transfer from FeII FeII binding, FeII redox potential and electronic matrix coupling of the FeII and the type 1 CuII (T1Cu) in the ferroxidase. We will do this by interconverting laccase and ferroxidase enzymes based on our design principles. In addition, we will test our hypothesis that another cohort of carboxylate side chains stabilizes the increasing negative charge on dioxygen as it is reduced in 2, 2e- steps at the trinuclear cluster (TNC). Last we will test our hypothesis that the coordination changes associated with O2 reduction at the TNC trigger e- transfer from the T1 Cu via the canonical MCO His-Cys-His motif that connects the two. In Aim II we propose to test our hypothesis that Fox1 in Chlamydomonas reinhardtii (Cr) is a human ceruloplasmin-like ferroxidase. We will characterize the Fox1 protein, demonstrating that it has the ferroxidase-specificity motifs that support both FeII oxidation and FeIII trafficking in a Fox1, Ftr1 complex in the Cr PM. We will construct mutants of both Fox1 and Ftr1 that we predict will be sensitive to a FeIII-chelator acting as a metabolite trap in Fe-trafficking between the two proteins in Fe-uptake; we have used this classic test of channeling in the Sc Fet3, Ftr1p complex. We propose that the Fe-trafficking between Fox1 and Ftr1 in Cr provides a realistic model of the putative FeIII-trafficking between hCp and transferrin. In Aim III, we will test our hypothesis that the two human fungal pathogens, Candida albicans and Cryptococcus neoformans express an equivalent PM Fet, Ftr high-affinity Fe-uptake complex. We will quantify the 59Fe-uptake kinetics via these complexes both in situ and in recombinant form in Sc. Targeting specific ferroxidase and Fe-trafficking residues in the Ca and Cn proteins, we will test our hypothesis that these mutants exhibit the channeling defect exhibited by the Sc homologues. We propose that strains expressing these channeling mutants will exhibit a reduced virulence in vitro and in vivo. Last, using these chelator-sensitive clones, the NCI diversity collection will be mined for compounds with the potential as inhibitors of Fet, Ftr Fe-uptake in these fungal pathogens. All oxygen-utilizing organisms from fungi to humans require the activity of a copper oxidase enzyme - a multicopper oxidase - to manage their metabolism of the essential nutrient, iron. Fungi from baker´s yeast to the human pathogens C. albicans and C. neoformans use these enzymes to acquire the iron they need to thrive and survive. The goal of this research is to take a snap-shot of how these enzymes work and then to use this knowledge to block their trafficking of iron as way of suppressing the virulence of pathogenic fungi in both otherwise healthy and immunocompromised patients
Keywords: Address; Aerobiosis; Affinity; Archives; autooxidation; base; Binding (Molecular Function); Biology; Candida albicans; carboxylate; Cell membrane; Ceruloplasmin; Charge; Chelating Agents; Chlamydomonas reinhardtii; cohort; Collection; Complex; copper oxidase; Coupled; Coupling; Cryptococcus neoformans; cytotoxicity; Databases; Defect; design; Dioxygen; electron donor; Electron Transport; Electronics; Electrons; Environment; Enzymes; Eukaryota; Exhibits; Family; fitness; fungus; Generations; Genome; Goals; Health; Homeostasis; Homologous Gene; Human; Hydrolysis; Immunocompromised Host; In Situ; In Vitro; in vivo; inhibitor/antagonist; insight; Ions; Iron; Kinetics; Knowledge; Laccase; Maps; Membrane; Metabolic Pathway; Metabolism; Metals; Mining; Modeling; mutant; Nutrient; Organism; Oxidases; oxidation; Oxidation-Reduction; Oxygen; pathogen; Pathway interactions; permease; Physiology; Plants; Play; Positioning Attribute; prevent; Process; programs; Protein Conformation; Proteins; Protons; Reaction; Reactive Oxygen Species; Recombinants; Reducing Agents; repository; Research; Resistance; Role; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Shipping; Ships; Side; Solvents; Specificity; Stream; Structure; Structure-Activity Relationship; System; Testing; Thermodynamics; Time; trafficking; Transferrin; uptake; Virulence; Work; Yeasts
Project start date: 1999-05-01
Project end date: 2012-03-31
Budget start date: 1-APR-2011
Budget end date: 31-MAR-2012
PFA/PA: PA-07-070
5R01DK053820-12 (2011): $281802
ENHANCING THE SAFETY OF ALLOGENEIC TRANSPLANTATION
J Daniel
University Of Minnesota Twin Citiescity: Minneapolis country: United States (us)
Grant 2U10HL069290-11 from National Heart, Lung, And Blood Institute
Abstract: Multicenter collaboration to reduce the risks inherent in allogeneic hematopoietic cell transplantation (HCT) is essential to dampen the dangers of regimen toxicity, infection, transfusion dependence, graft vs. host disease (GVHD) and malignant relapse. Ongoing efforts to address all these risks have been a focus of Network participation for investigators at the University of Minnesota. Our commitment to the Network has included participation as Steering Committee chair, national PI of four protocols, leading the first four Network publications and committing institutional resources to develop and successfully execute Network trials. Because umbilical cord blood (UCB) transplantation is still limited by delayed or failed engraftment, we initiated studies to augment the homing and hematopoietic recovery of UCB grafts based on preclinical data suggesting that complement fragment C3a can accomplish these goals. An ongoing University of Minnesota phase 1 trial testing C3a priming of one of a pair of UCB units to augment engraftment and enhance predominance of the primed unit provides the clinical background for a proposed multicenter phase 11. We propose a protocol testing whether C3a priming of one UCB unit can increase the likelihood of that unit´s predominance as the engrafting hematopoietic source; can augment and accelerate multilineage hematopoietic recovery; and can enhance immune reconstitution thereby lessening risks of infection without excess graft vs. host disease. We offer our continued commitment to the Network´s success in developing new protocols, identifying patients suitable for Network trial enrollment and performing protocol specific procedures. We will try to exceed the Network´s standards in our commitment to enhance multicenter trial success and advance scientific progress within the Network. Multicenter collaborative trials are essential to formally test new ways to enhance the safety of allotransplantation and improve outcomes for our patients. PUBLIC RELEVANCE Improvement in the outcomes of allergenic transplantation requires control of early toxicity, risks of infection, transfusion dependence, graft vs. host disease and relapse. Multicenter trials within the BMT CTN have tackled all these areas and at the University of Minnesota we are committed to advancing the development and execution of high-quality Network trials. We propose augmenting the success of umbilical cord blood engraftment by priming one of a pair of UCB units with complement fragment C3a, to extend preliminary University of Minnesota data to a multicenter phase II trial. Improving engraftment and post transplant immune recovery may limit these hazards and augment the success of all transplantation
Keywords: Address; Advanced Development; Allogenic; Area; base; Cell Transplantation; Clinical; Collaborations; collaborative trial; Commit; Complement; Complement 3a; Data; Dependence; Disease; disorder risk; Engraftment; Enrollment; Goals; Graft-vs-Host Disease; hazard; Hematopoietic; Hemorrhage; Homing; Homologous Transplantation; Immune; improved; Infection; Malignant - descriptor; Minnesota; Multicenter Trials; Outcome; Pancytopenia; Patients; Phase; Phase I Clinical Trials; Phase II Clinical Trials; pre-clinical; Procedures; Protocols documentation; Publications; reconstitution; Recovery; Recurrent disease; Regimen; Relapse; Research Personnel; Resources; Risk; Safety; Scientific Advances and Accomplishments; Source; success; Testing; Toxic effect; Transfusion; Transplantation; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; Universities
Project start date: 2001-09-30
Project end date: 2017-06-30
Budget start date: 8-AUG-2011
Budget end date: 30-JUN-2012
PFA/PA: RFA-HL-11-013
2U10HL069290-11 (2011): $149490
IS INSOMNIA A RISK FACTOR FOR DECREASED INFLUENZA (E.G., H1N1) VACCINE RESPONSE?
J Daniel, Asst. Professor
University Of North Texascity: Denton country: United States (us)
Grant 1R15AI085558-01A2 from National Institute Of Allergy And Infectious Diseases
Abstract: Influenza and insomnia are significant public health problems. The 2009 H1N1 pandemic had greater clinical impact on children and young adults; prompting the CDC to issue an H1N1 Infections Alert for Institutions of Higher Education. Although the influenza vaccine is recommended for everyone, the vaccination is ineffective in a modest percent of cases. One possibility is that concomitant health problems (e.g., insomnia) serve as stressors which can reduce the effectiveness of the influenza vaccination via suppressed antibody responses to the viral strains. Insomnia plays an integral role in host defense and development of inflammatory diseases, and is regulated by and may be altered by a number of immune messengers (e.g., cytokines). These insomnia- immune relationships need translation to the more clinically relevant area of vaccination response. Considering the prevalence of insomnia (46%-69% of primary care patients; 10-15% of college students), it is critical to determine if insomnia is a risk factor for reduced response to the influenza and other vaccinations, while controlling for confounding medical conditions and medications. Major theories suggest that once insomnia becomes chronic, it becomes a major stressor, and is no longer a result of stress. If this is true, then it would follow that insomnia would result in reduced effectiveness of the influenza vaccination via suppressed antibody responses to the viral strains. The proposed R15 (AREA) pilot study moves beyond the paradigm of measuring isolated immune factors to a more integrated, systemic view of immune function, and is the first exploration of in vivo immune response in a healthy young adult insomnia population. The primary aim of the proposed study is to determine if insomnia is a risk factor for lower influenza vaccine antibody response. To test this aim, a repeated-measures between subjects design will be used to assess groups differences (people with insomnia vs. people without insomnia) pre-vaccination and 4 weeks post-vaccination on measures of antibody titers for the specific influenza strains (including H1N1) used in the yearly vaccine. Insomnia diagnoses will be determined with clinical interview. Antibody amounts, antibody sub-class and quality will be measured utilizing hemagglutination inhibition (HI), serum anti-influenza IgG and IgG1, and a virus neutralization assay. Recruitment will occur over 2 years/influenza seasons (32 subjects per group each year), with a final sample size of N=128. It is likely that each yearly vaccine will be comprised of different viral strains (determined yearly by the World Health Organization). The current design will allow assessment of antibody response to the influenza vaccine as a whole (i.e., including all possible virus strains over 2 years) using MANOVA, as well as each individual virus strain per year using follow-up ANOVA. All immune assays (HI, ELISA, viral neutralization) will be performed at the Iowa State University Health and Human Performance research laboratory. Additional analyses will examine a meditational role between insomnia and stress, as well as determining if other measures of sleep disturbance are better predictors of antibody response. Influenza and other communicable diseases pose a significant health risk to the American public. Vaccines are available for many of these diseases, but they are not always effective. Insomnia is a highly prevalent disease that may suppress immune functioning and reduce the effectiveness of these vaccines. If evidence supports this, then treating insomnia may become an important public health initiative to help improve the prevention of communicable diseases
Keywords: 0-11 years old; Active Follow-up; adult youth; advanced age; Affect; Aged 65 and Over; American; anti-flu; anti-influenza; Antibodies; antibody biosynthesis; Antibody Formation; Antibody Production; Antibody Response; antiflu; Area; Assay; B cell differentiation factor; B cell stimulating factor 2; B-Cell Differentiation Factor-2; B-Cell Stimulatory Factor-2; BCDF; Bioassay; Biologic Assays; Biological Assay; Blood Serum; BSF-2; BSF2; BSF2 (B cell stimulating factor 2); CDC; Centers for Disease Control; Centers for Disease Control (U.S.); Centers for Disease Control and Prevention; Centers for Disease Control and Prevention (U.S.); Child; Child Youth; children; Children (0-21); Chronic; Chronic Insomnia; Clinical; clinical relevance; clinical significance; clinically relevant; clinically significant; college student; Communicable Diseases; cytokine; Cytotoxic cell; design; designing; Development; Diagnosis; Differentiation Factor, B-Cell; Disease; Disease Outbreaks; disease/disorder; Disorder; drug/agent; Drugs; Education; Educational aspects; Effectiveness; Elderly; Elderly, over 65; elders; ELISA; Enzyme-Linked Immunosorbent Assay; flu immunisation; flu infection; Flu vaccination; Flu vaccine; follow-up; Gamma Globulin, 7S; geriatric; Grant; Grippe; H1N1 vaccine; H1N1 Virus; Health; Hemagglutination; Hepatitis B; Hepatitis B Infection; Hepatocyte-Stimulating Factor; Host Defense; host response; HPGF; Human; Human, Child; Human, General; Hybridoma Growth Factor; IFN-beta 2; IFNB2; IgG; IgG1; IL-6; IL6 Protein; Immune; immune function; Immune Function, Cellular; Immune response; immunoglobulin biosynthesis; Immunoglobulin G; immunoresponse; improved; in vivo; Individual; Infection; Infectious Disease Pathway; Infectious Diseases; Infectious Diseases and Manifestations; Infectious Disorder; Inflammatory; Influenza; Influenza A Virus, H1N1 Subtype; Influenza immunization; influenza infection; Influenza vaccination; Influenza Vaccines; Influenza Virus; Influenza virus vaccine; influenzavirus; influenzavirus (unspecified); Insomnia; Insomnia Disorder; Institution; interferon beta 2; Interleukin 6 (Interferon, Beta 2); Interleukin-6; Intervention; Intervention Strategies; interventional strategy; Interview; Investigation; Investigators; Iowa; K lymphocyte; Laboratory Research; late life; later life; Man (Taxonomy); Man, Modern; Measures; Medical; Medication; MGI-2; Myeloid Differentiation-Inducing Protein; Natural Killer Cells; NK Cells; older adult; older person; Outbreaks; pandemic; pandemic disease; pathway; Pathway interactions; Patients; Performance; Pharmaceutic Preparations; Pharmaceutical Preparations; Pilot Projects; pilot study; Plasmacytoma Growth Factor; Play; Population; Prevalence; Prevention; Primary Care; Primary Health Care; Primary Healthcare; Prophylactic vaccination against influenza; Public Health; public health medicine (field); public health relevance; Research Personnel; Researchers; response; Risk; Risk Factors; Role; Sample Size; Sampling; Seasons; senior citizen; Serum; serum hepatitis; Sleep; Sleep disturbances; Sleeplessness; social; social role; Stress; stressor; Testing; theories; Translations; United States; United States Centers for Disease Control; United States Centers for Disease Control and Prevention; Universities; university student; Vaccinated; Vaccination; vaccine effectiveness; Vaccines; Viral; Viral Hepatitis B; Virus; Viruses, General; WHO; World Health Organization; young adult; youngster
Relevance: NARRATIVE Influenza and other communicable diseases pose a significant health risk to the American public. Vaccines are available for many of these diseases, but they are not always effective. Insomnia is a highly prevalent disease that may suppress immune functioning and reduce the effectiveness of these vaccines. If evidence supports this, then treating insomnia may become an important public health initiative to help improve the prevention of communicable diseases
Project start date: 2011-02-15
Project end date: 2014-01-31
Budget start date: 15-FEB-2011
Budget end date: 31-JAN-2014
PFA/PA: PA-10-070
1R15AI085558-01A2 (2011): $442838
CONTROL OF REGULATORY T CELL HOMEOSTASIS AND FUNCTION BY THE TH1
J Daniel
Benaroya Research Inst At Virginia Masoncity: Seattle country: United States (us)
Abstract: Immune-mediated infiammatory diseases are a major public health issue. In the lungs, these can be triggered by exposure to environmental anfigens and toxins, or by infection, and can result in severe respiratory disease and death. Defining the immunoregulatory mechanisms that normally function to prevent pulmonary inflammafion is therefore key to understanding the efiology of these diseases, and for developing therapeutic strategies to boost these acfivities in pafients. Regulatory T cells (TR) expressing the transcripfion factor Foxp3 play a critical role in prevenfing autoimmunity and limifing immune-mediated inflammafion. We have shown that during type-1 inflammatory responses, Foxp3+ TR upregulate the Thlspecifying transcripfion factor Tbx21 (T-bet), and that T-bet expression is crifical for proper TR homeostasis and funcfion during Thi-mediated inflammation. Therefore, the goals of this proposal are to determine in detail how loss of T-bet specifically within Foxp3+ TR impacts the initiafion, progression and terminafion of Thi responses in models of acute and persistent lung infection in vivo (Specific Aim 1), define the cytokines and cellular signals that direct TR expression of T-bet (Specific Aim 2), and analyze at the molecular level how FoxpS and T-bet combine to control the expression of genes involved in JhlfTR differenfiafion, homeostasis and funcfion (Specific Aim 3). Together, these experiments will generate an unprecedented understanding of the molecular specializafion of TR subsets during type-1 inflammafion, and provide a new framework in which to understand how so-called ´master transcription factors´ direct the funcfional differenfiation of CD4+ T cell subsets
Keywords: Acute; Address; Adoptive Transfer; Antigen-Presenting Cells; Antigens; Autoimmunity; base; CD4 Positive T Lymphocytes; cell type; Cells; Cessation of life; chemokine receptor; Chronic; Chronic Obstructive Airway Disease; clinical application; CXCR3 gene; cytokine; Disease; Etiology; Exposure to; Extracellular Matrix; Gene Expression; Genes; Goals; Granulomatous; Homeostasis; Immune; Immune response; in vivo; Infection; Inflammation; Inflammatory; Inflammatory Response; Interferons; Leukocytes; Lung; Lung diseases; Lymphoid; Mediating; microbial; Modeling; Molecular; Mus; Mycobacterium tuberculosis; Patients; Peripheral; Play; Pneumonia; prevent; public health medicine (field); Regulation; Regulatory T-Lymphocyte; research study; response; Role; Signal Transduction; Site; STAT1 gene; Stimulus; T-Lymphocyte Subsets; Testing; Th1 Cells; Therapeutic; TNFRSF5 gene; Toxin; transcription factor; Tuberculosis; Work
Relevance: Understanding how TR modulate Thi-mediated immune responses has clear and direct implicafions in the clinical application of TR for the treatment of immunolgic pulmonary diseases caused by dysregulated Th1 cell responses, such as granulomatous infiammation associated with persistent Mycobacterium tuberculosis infecfion, hypersensifivity pneumonitis, and chronic obstructive pulmonary disease
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
5P01HL098067-02_6080 (2011): $280641
FUNCTIONAL SPECIALIZATION OF FOXP3+ REGULATORY T CELLS
J Daniel
Benaroya Research Inst At Virginia Masoncity: Seattle country: United States (us)
Grant 5R01AI085130-02 from National Institute Of Allergy And Infectious Diseases
Abstract: Immune-mediated autoimmune and inflammatory diseases are a major public health issue. Defining the regulatory mechanisms that normally function to prevent pulmonary inflammation is therefore a key to understanding the etiology of these diseases, and for developing therapeutic strategies to boost these activities in patients. Regulatory T cells (TR) expressing the transcription factor Foxp3 play a critical role in preventing autoimmunity and limiting immune-mediated inflammation. We have shown that during type-1 inflammatory responses, Foxp3+ TR upregulate the Th1-specifying transcription factor Tbx21 (T-bet), and that T-bet expression is critical for proper TR homeostasis and function during Th1-mediated inflammation. Therefore, the goals of this proposal are to determine in detail how loss of T-bet specifically within Foxp3+ TR impacts the initiation, progression and termination of Th1 responses in vivo (Specific Aim 1); analyze at the molecular level how Foxp3 and T-bet combine to control the expression of genes involved in Th1/TR differentiation, homeostasis and function (Specific Aim 2); and to identify the cytokines and cellular signals that control the phenotypic and functional differentiation of different TR subsets (Specific Aim 3). Understanding how regulatory T cells modulate Th1- and Th17mediated immune responses has clear and direct implications in the clinical application of these cells for the treatment of immune-mediated inflammatory and autoimmune diseases caused by dysregulated Th1 cell responses, such as granulomatous inflammation associated with persistent Mycobacterium tuberculosis infection, hypersensitivity pneumonitis, psoriasis, rheumatoid arthritis, type-1 diabetes and multiple sclerosis
Keywords: Address; Adopted; Adoptive Transfer; Antigen-Presenting Cells; Autoimmune Diseases; Autoimmune Process; Autoimmunity; base; CD4 Positive T Lymphocytes; cell motility; Cell physiology; cell type; Cells; chemokine receptor; Chronic; clinical application; CXCR3 gene; cytokine; Development; Disease; Etiology; Extrinsic allergic alveolitis; Gene Expression; Genes; Goals; Granulomatous; Helper-Inducer T-Lymphocyte; Homeostasis; Immune; Immune response; in vivo; Infection; Inflammation; Inflammatory; Inflammatory Response; Insulin-Dependent Diabetes Mellitus; Lung; Lymphoid Tissue; Mediating; microbial; Molecular; Multiple Sclerosis; Mus; Mycobacterium tuberculosis; Patients; Peripheral; Phenotype; Play; Pneumonia; Population; prevent; Production; programs; Psoriasis; public health medicine (field); public health relevance; Regulatory T-Lymphocyte; response; Rheumatoid Arthritis; Role; Signal Transduction; Site; Specific qualifier value; Stimulus; T-Lymphocyte; Testing; Th1 Cells; Therapeutic; TNFRSF5 gene; transcription factor; Tuberculosis; Work
Relevance: Understanding how regulatory T cells modulate Th1- and Th17mediated immune responses has clear and direct implications in the clinical application of these cells for the treatment of immune-mediated inflammatory and autoimmune diseases caused by dysregulated Th1 cell responses, such as granulomatous inflammation associated with persistent Mycobacterium tuberculosis infection, hypersensitivity pneumonitis, psoriasis, rheumatoid arthritis, type-1 diabetes and multiple sclerosis
Project start date: 2010-06-01
Project end date: 2015-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-07-070
5R01AI085130-02 (2011): $427507
THE NEUROIMMUNOLOGY OF VIRAL INFECTION
J Daniel
University Of Oklahoma Hlth Sciences Ctrcity: Oklahoma City country: United States (us)
Grant 2R01AI053108-08 from National Institute Of Allergy And Infectious Diseases
Abstract: Herpes simplex virus type 1 (HSV-1) is one of the most common neurotropic pathogens in humans with a seroprevalence rate increasing up to 60-80% by the 5th decade of life. Clinical diseases associated with HSV-1 include herpetic stromal keratits, the leading cause of infectious corneal blindness in the industrialized world, and frank sporadic encephalitis, a rare but debilitating disease that can result in death. Resistance to infection includes the innate immune system. Central to the innate immune response to virus infection is the type I interferon (IFN) pathway which upon activation by a variety of means including pattern recognition receptors results in the expression of potent anti-viral pathways that block virus replication at the transcriptional and translational levels. Recently, we have begun to explore the relationship between virus infection in the nervous system (peripheral and central) and the role of the type I IFN system in local host resistance and the ensuing adaptive immune response to the insult. Mice deficient in a functional type I IFN pathway (i.e., absence of the alpha chain of the type I IFN receptor, CD118-/-) are found to be highly sensitive to HSV-1 succumbing to infection within 6 days associated with an elevation in virus titer in the peripheral and central nervous systems. Moreover, there is a massive loss of cells residing in the draining lymph node associated with an increase in T cell apoptosis and necrosis. There are also notable changes in recruitment of leukocytes residing in the nervous system post infection of CD118-/- mice. Changes in leukocyte recruitment to the nervous system are reflected by specific changes in select chemokines including CXCL1, CXCL10 and CCL2 in the infected tissue. However, the contribution of resident cells (including astrocytes and microglia) versus infiltrating leukocyte populations within the nervous system or the organized lymphoid tissue (i.e., draining lymph node) in the context of a functional type I IFN pathway in response to infection and the ensuing adaptive immune response and neuropathology is unexplored. We propose to employ mice deficient in select genes as well as mouse chimeras to address this question. Preliminary, unpublished data suggest the resident cells residing in the brain stem with an intact type I IFN system are critical to control viral infection whereas in the peripheral nervous system (i.e., trigeminal ganglion) or cornea, both resident and bone marrow-derived cells with a functional type I IFN pathway are required to maximize resistance to infection. We propose two specific aims within this competitive renewal. The first aim will test the hypothesis microglia with a functional type I IFN receptor maintain resistance to HSV-1 following ocular infection. The second aim will test the hypothesis that both leukocytes and resident cells within the TG, cornea, and draining lymph node with a functional type I IFN receptor are required to maximize resistance to HSV-1 infection. It is anticipated the outcome of this project will identify key central components of innate and adaptive immunity within the CNS and peripheral tissues (including sensory ganglia and cornea) that are critical for viral surveillance unique to each tissue. In so doing, a focus on the development of vaccines that utilize (via expansion or activation) such components will provide superior efficacy compared to current strategies that utilize viral encoded proteins that typically generate an immune response incorporating only antibody production or T effector cells. Alternatively, mediators of neuropathogenesis can be targeted to alleviate collateral damage and thus, preservation of neurons in patients that suffer from acute or recurrent HSV-1 CNS or peripheral nerve infection. Herpes simplex virus type 1 (HSV-1) is a human pathogen that can lead to sporadic encephalitis or other neuropathological conditions in patients that suffer reactivation episodes. The role of the principal innate immune response associated with the control of viral replication and spread, the type I interferons (IFNs), as it relates to the reduction of CNS inflammation and development of the adaptive immune response within the nervous system will be investigated using whole animal, cellular, and molecular techniques. It is anticipated the outcome of the study will identify novel mediators of inflammation responsible for tissue destruction and neuropathology but irrelevant in the control of virus infection. Consequently, by targeting the inflammatory mediators, it may be possible to contain the severity of neuroinflammation without compromising the host ability to control virus replication and spread
Keywords: Acute; adaptive immunity; Address; Animals; anterograde transport; Antibody Formation; Apoptosis; Astrocytes; base; Biological Preservation; Blindness; Bone Marrow; Brain Stem; CCL2 gene; Cells; Cessation of life; chemokine; Chimera organism; Clinical; comparative efficacy; Cornea; CXCL1 gene; CXCL10 gene; Data; defined contribution; Development; Disease; Effector Cell; Encephalitis; Epithelium; Eye Infections; Fiber; Genes; Herpesvirus 1, Human; Herpetic Keratitis; Host resistance; Human; Immune response; Immune system; Immunohistochemistry; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory Response; Interferon Type I; Lead; Leukocytes; Life; lymph nodes; Lymphoid Tissue; Magnetic Resonance Imaging; Mediator of activation protein; Microglia; Molecular; Mus; Natural Immunity; Necrosis; Nervous system structure; Neuraxis; neuroimmunology; neuroinflammation; Neurons; Neuropathogenesis; neuropathology; neurotropic; novel; Outcome; Outcome Study; pathogen; Pathology; Pathway interactions; Patients; Pattern recognition receptor; Peripheral; Peripheral Nerves; Peripheral Nervous System; Population; Proteins; Recurrence; Relative (related person); Resistance; Resistance to infection; response; Role; Sensory; Sensory Ganglia; Seroprevalences; Severities; Structure of trigeminal ganglion; System; T-Lymphocyte; Techniques; Testing; Tissues; Toll-like receptors; type I interferon receptor; vaccine development; Viral; Viral Load result; Virion; Virus; Virus Diseases; Virus Replication
Relevance: Herpes simplex virus type 1 (HSV-1) is a human pathogen that can lead to sporadic encephalitis or other neuropathological conditions in patients that suffer reactivation episodes. The role of the principal innate immune response associated with the control of viral replication and spread, the type I interferons (IFNs), as it relates to the reduction of CNS inflammation and development of the adaptive immune response within the nervous system will be investigated using whole animal, cellular, and molecular techniques. It is anticipated the outcome of the study will identify novel mediators of inflammation responsible for tissue destruction and neuropathology but irrelevant in the control of virus infection. Consequently, by targeting the inflammatory mediators, it may be possible to contain the severity of neuroinflammation without compromising the host ability to control virus replication and spread
Project start date: 2003-04-01
Project end date: 2016-03-31
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-10-067
2R01AI053108-08 (2011): $367939
5R01AI053108-07 (2009): $279097
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