Daniel Kelly - US grants

A Physics Laboratory Course improvement program has been developed by Physics faculty at Southwestern Oregon Community College to more closely align laboratory exercises and General Physics and Engineering Physics lecture topics for science and non science majors who transfer to university level programs. The principal goal is to increase opportunities for students to perform a wider variety of experiments using appropriate laboratory equipment and to enhance learning through physics simulations using microcomputers. The focus will be on single topic labs keyed to Physics lecture material. A commitment to new equipment, including computers, will update and enhance existing laboratory equipment. Computer simulations of various physics experiments enhance understanding of more complex general physics concepts which are not possible to investigate in the laboratory - such as the motion of many bodies under gravity, or electric fields in complicated geometrics - while introducing students to computer use in a scientific setting. The college with match the award with an equal amount of funds.

Medium-chain acyl-CoA dehydrogenase (MCAD,EC 1.3.99.3) is a mitochondrial flavoenzyme which catalyzes the initial, rate-limiting, reaction in fatty acid beta-oxidation. Inherited MCAD deficiency, a cause of fasting coma, liver dysfunction, and sudden death in childhood, reflects the importance of this enzyme in energy metabolism. The MCAD gene is highly regulated, in parallel with fatty acid oxidation rates, among tissues and during development. Elucidation of the mechanisms involved in the regulation of the MCAD gene will provide insights relevant to MCAD deficiency and to the understanding of mechanisms involved in the regulation of expression of nuclear genes encoding mitochondrial enzymes involved in fatty acid metabolism under normal conditions and in a variety of physiologic and disease states. The major goals of this proposal include the structural and functional characterization of the promoter and upstream cis-acting regulatory regions of the human MCAD gene including identification of the elements involved in tissue-specific, developmental, and retinoic acid-responsive transcriptional regulation. Ultimately, we hope to identify regulatory sequences and transacting regulatory DNA binding proteins involved in coordinate control of genes encoding metabolic enzymes and mitochondrial proteins. The promoter region and regulatory elements will be characterized by transfecting a variety of chimeric plasmids containing varying lengths of MCAD gene 5'-flanking DNA fused to the bacterial chloramphenicol acetyltransferase (CAT) gene into mammalian cells derived from several tissues with differing MCAD mRNA expression levels. Retinoic acid response elements will be localized by identifying the regulatory sequences of the MCAD gene which confer retinoic acid-responsive transcriptional activation to the MCAD-CAT plasmids in mammalian cells in culture. The regulatory sequences will be compared to sequences of known regulatory elements in other genes, particularly those encoding enzymes involved in metabolism. The regulatory elements will be further characterized by performing DNA-nuclear protein binding assays. Ultimately, the trans-acting regulatory proteins involved in the transcriptional regulation of the MCAD gene will be identified by isolating and characterizing their cDNAs. The role of the regulatory elements in developmental and tissue-specific regulation of the MCAD gene in vivo will be evaluated by employing a transgenic mouse system.

Medium-chain acyl-CoA dehydrogenase (MCAD), a nuclear encoded mitochondrial enzyme, catalyzes the initial step in the fatty acid beta- oxidation cycle. The importance of this enzyme in cellular energy metabolism is underscored by the severe and often fatal consequences of inherited human MCAD deficiency. Expression of the MCAD gene is highly regulated in parallel with fatty acid oxidation rates among tissues, during development and in response to fasting and physiologic stimuli. This competitive renewal is designed to test the hypothesis that human MCAD gene promoter sequences identified in vitro, including several novel nuclear hormone receptor response elements and Sp1 binding sites, confer transcriptional regulation of MCAD gene expression in vivo. This hypothesis will be tested by comparing the transcriptional activity of human MCAD gene promoter fragments containing point mutations targeted to known regulatory elements with that of the wild-type promoter in transgenic mice. Elements involved in tissue- and developmental-stage specific expression of the MCAD gene will be delineated. Following localization, the tissue and developmental regulatory elements will be evaluated upstream of a minimal promoter in transgenic mice. The transgenic mice will also be used to identify and localize cis-acting elements that confer regulation of MCAD gene expression in response to fasting, perturbations in mitochondrial fatty acid beta-oxidation, and chronic dietary changes. Transgenic mice containing the fasting and metabolic response elements upstream of heterologous promoters will be produced and characterized as an initial step in the production of tissue- restricted, expression systems responsive to dietary and physiologic stimuli known to increase cellular demands. Lastly, cDNAs encoding the transcription factors that bind the MCAD gene fasting responsive elements will be cloned and characterized. This work should lead to an improved understanding of the pathogenesis of inborn errors in fatty acid oxidation and the development of expression strategies that should be useful in the study and treatment of a variety of inborn and acquired diseases due to abnormalities in cellular energy metabolism.

The aim of this research is to evaluate myocardial fatty acid oxidation in comparison with mitochondrial oxidation using positron emission tomography with 11C-palmitate and 11C-acetate, respectively, in patients with medium- and long chain acyl-CoA dehydrogenase (MCAD and LCAD) deficiency and compare results to those obtained in normal volunteers. The long term goal is to evaluate whether the myocardial manifestations of MCAD and LCAD deficiency can be evaluated with noninvasive interrogation of myocardial metabolism.

DESCRIPTION (provided by applicant): Cardiac hypertrophy is a common condition often predisposing to heart failure and sudden death. Despite recent progress in the identification of cellular signaling pathways involved in cardiac hypertrophic growth, little is known about the downstream molecular events that lead to contractile dysfunction in the hypertrophied heart. The expression of enzymes involved in mitochondrial fatty acid oxidation (FAO), the chief cardiac energy source, is coordinately downregulated in acquired forms of pathologic cardiac hypertrophy. We have recently demonstrated that this metabolic regulatory switch is due to deactivation of a transcriptional regulatory complex comprised of the nuclear receptor, PPARalpha, and its cardiac-enriched, coactivator PGC- 1. This renewal proposal is designed to test the hypothesis that the activity of PPARalpha/PGC-l plays a critical role in determining whether the hypertrophied heart becomes dysfunctional (pathologic remodeling) or maintains normal function as with physiologic forms of hypertrophy. We propose that the activity of PPARalpha is influenced by a balance of upstream signaling pathways linked to growth stimuli. The experiments planned for Specific Aim 1 are designed to characterize the upstream signaling events controlling the activity of the PPARalpha/PGC-1 complex and its target genes involved in mitochondrial metabolism during hypertrophic growth. The goal of Specific Aim 2 will be to evaluate the regulation of this energy metabolic pathway during physiologic hypertrophic growth due to exercise training. The objective of Specific Aim 3 is to systematically evaluate the effects of PPARalpha and PGC-1 in pathologic forms of cardiac hypertrophy using gain-of-function and loss-of-function genetic strategies in mice. The cardiac phenotype of these mice will be evaluated using molecular biologic, physiologic, and metabolic endpoints. Experiments proposed in Specific Aim 4 will evaluate the cardiac metabolic and functional response of mice lacking PPARalpha or PGC- 1 to exercise training. The longterm objective of this project is to identify molecular targets for novel therapeutic strategies aimed at preventing the pathologic metabolic remodeling of the hypertrophied heart.

Inherited defects in cardiac lipid metabolism are an important cause of inherited cardiomyopathy in sudden death in children and young adults. Recent studies also indicated that similar abnormalities in myocardial fatty acid utilization due to alterations in cardiac fatty acid utilization is poorly understood but may involve accumulation of toxic lipid intermediates within the cardiac myocyte. To investigate the molecular pathogenesis of cardiomyopathy due to abnormalities in cardiac lipid metabolism, we have begun to establish and characterize genetically engineered mouse lines with altered expression of enzymes and proteins involved in myocardial fatty acid import and utilization. Characterization of the cardiac phenotype of these mouse lines will allow us to develop murine models of metabolic cardiomyopathy and to test the hypothesis that accumulate of intracellular lipid plays a role in the genesis of heart failure and sudden death. The aims of this proposal include; 1) the development and characterization of mice with reduced expression of cardiac fatty acid oxidation (FAO) by altering the activity of transcription factors known to control the expression of this pathway to heart, 2) to produce and characterize mice with constitutively increased long-chain fatty acyl-CoA synthetase (FACS) in heart, 3) to characterize the cardiac phenotype of mice with diminished capacity for fatty acid utilization and/or increased fatty acid import under dietary and metabolic conditions known to precipitate heart failure in humans with inborn efforts in FAO, and 4) to characterize the cardiac phenotype of the mouse lines in response to physiologic conditions known to increase myocardial fatty acid utilization requirements. The results of these studies should provide significant insight into the role of perturbations in myocardial lipid metabolism and the pathogenesis of inherited and acquired forms of heart failure.

Significant advances in the development of genetically engineered mice lacking or over-expressing specific proteins has proven to be a powerful research tool. As described in this SCOR proposal, a number of interesting transgenic and gene "knockout" models will be developed to explore the role of specific enzymes and proteins in the physiology and pathophysiology of the cardiovascular system. The mouse cardiovascular Physiology Core will assist with the characterization of the mouse models. This Core will provide rigorous physiologic evaluation of mouse models including; 1) characterization of the structure and function of the heart and great vessels using trans-thoracic echocardiography, 2) cardiovascular hemodynamic measurements of the mouse circulatory system using open- and closed-chest cardiac catheterization, 3) identification and characterization of cardiac rhythm disturbances in mice using ambulatory telemetric electrocardiographic monitoring, and 4) to evaluate the responses to short-term and long-term exercise studies in mice. The Mouse Cardiovascular Physiology Core will accomplish these goals in dedicated space within the Clinical Sciences Research Building at Washington University School of Medicine. It is anticipated after the evaluation of the mouse models developed in this proposal will identify cardiovascular abnormalities relevant to the pathogenesis of pediatric cardiovascular disease.

This is a new application to acquire a digital high frequency cardiovascular ultrasound system to be used for cardiac imaging in small animals, mainly mice, and studies of vascular structure and function in humans. This system will be shared among a scientifically diverse group of PHS-supported investigators representing 7 departments at Washington University (Internal Medicine, Pediatrics, Pathology, Molecular Biology & Pharmacology, Cell Biology & Physiology, Biomedical Engineering, and Physics). Each member of the group has successfully used this system, which is available on a temporary basis as a loaner, and has demonstrated a significant need in the future. Accordingly, this requested shared instrument will have a major impact on the success of current and suture PHS-supported research activities of this user group. The proposed applications of the cardiovascular ultrasound system are focused on three major target research areas: i) characterization of mouse models of cardiomyopathy due to alterations in myocardiac hypertrophy in vivo using mouse models, and iii) application of high resolution ultrasound to vascular structure and functions in humans. In addition, several users will conduct basic ultrasonic research relevant to the physiology and material properties of the mouse models of cardiovascular disease developed by the user group. The requested instrumentation will be housed in dedicated space within the Small Animal Cardiovascular Physiology and Imaging Core Facility in the Center for Cardiovascular Research at Washington University School of Medicine. The system will be maintained and operated by a technical director and co-director within the Core facility. Validation studies are currently underway and plans have been made to establish an echocardiography database. This outstanding imaging tool will promote the success of the individual user's research program as well as facilitate multi- disciplinary efforts aimed at the characterization of the cardiovascular phenotype of mice generated through gene targeting and transgenic strategies. Acquisition of the requested cardiovascular ultrasound system is pivotal for the success of current and future PHS-funded cardiovascular research programs at Washington University.

The long term objective of this Pediatric SCOR is to determine the molecular bases of defective human cardiac morphogenesis and myocardial function which results in congenital heart disease (CHD) and pediatric cardiomyopathy (CM). The underlying hypothesis is that single gene defects at multiple loci in the human genome cause most pediatric cardiac structural and myopathic diseases. Two corollaries will also be explored, that (i) genetic abnormalities at the same locus have variable expressivity and can result in different phenotypes and (ii) genotype- phenotype correlations exist. A multi-disciplinary approach encompassing 14 investigators, 6 clinical and laboratory projects, and 4 core units at three locations is proposed. Molecular genetic studies of a St. Louis family with dominantly-inherited dilated CM; CM or sudden death secondary to mutations in mitochondrial fatty acid oxidation enzymes; patients with CHD associated with at the human at the human 8p23 locus encompassing the GATA-4 gene, a critical transcription factor in heart; elastin in heart; elastin mutations in supravalvar aortic stenosis; and dominantly-inherited atrial septal defects (ASD) mapped to 5p and 5q are the clinical focus. Mouse gene ablation and transgenic models to delineate (i) the pathogenesis of CM and sudden death in fatty acid oxidation defects, (ii) the essential role and downstream targets of GATA-4 expressed in mouse embryonic endoderm for mesoderm-mediated and downstream morphogenesis, (iii) the mechanisms by which elastin mutations disrupt elastic fiber assembly and vasculogenesis, and (iv) the mechanisms by which mutant mouse homologs of genes defective in human familial ASD result in CHD will be created. The molecular role of syndecans in determination of left-right asymmetry will be studied in the uniquely-manipulable Xenopus embryo system as a model for mechanisms underlying human heterotaxy syndromes and CHD. Characterization of defective human genes causing pediatric heart disease and the mechanisms through which mutations alter cardiac development and myocardial function is necessary before manipulations to prevent CHD and inherited CM are feasible.

DESCRIPTION (provided by applicant): We are witnessing an emerging epidemic of obesity and type 2 diabetes. The peroxisome proliferators-activated receptors (PPARs) are a family of nuclear receptors that control the expression of genes involved in cellular fatty acid metabolism. Using genetically-modified mice, we have recently unveiled exciting links between the muscle PPARalpha pathway and obesity-related insulin resistance. PPARalpha deficiency protects against insulin resistance and diabetes despite the development of an obese phenotype. Conversely, transgenic mice with muscle-specific overexpression of PPARalpha develop diet-induced glucose intolerance and insulin resistance despite maintaining a lean phenotype. This renewal proposal is designed to test the hypothesis that in states of caloric excess, re-direction of lipid to extra-adipose tissues, such as skeletal muscle, triggers PPARalpha-driven increases in mitochondrial fatty acid oxidation and reciprocal reduction in glucose utilization through gene regulatory mechanisms independent of the insulin signaling machinery. It is also proposed that over the long-term, increased fatty acid flux through oxidative pathways leads to muscle mitochondrial dysfunction. The objectives of this proposal will be achieved using genetically-modified mice and UCP-DTA mice, a murine model of Metabolic Syndrome and type 2 diabetes. Experiments proposed in Specific Aim 1 are designed to characterize the molecular regulatory mechanisms involved in the glucose intolerance and insulin resistance related to PPARalpha-driven increases in skeletal muscle fatty acid oxidation. The goal of Specific Aim 2 is to compare the effects of PPARbeta/delta and PPARalpha on gene regulation, muscle metabolism, obesity, and insulin resistance. Specific Aim 3 is designed to characterize the effects of the PPAR coactivators, PGC-1alpha and PGC-1beta, on muscle metabolism, and insulin resistance. Specific Aim 4 involves a series of intervention experiments aimed at modulating PPARalpha-driven skeletal muscle metabolic derangements. In the short-term we seek to develop a clear conceptual framework for the interaction between derangements in muscle lipid metabolism, and the development of diabetes. A long-term goal is to identify novel therapeutic targets relevant to Metabolic Syndrome.

Cardiac dysfunction is a common and important manifestation of diabetes mellitus. It is well recognized that cardiomyopathy occurs frequently in diabetic patients in the absence of known cardiac risk factors. Although little is known about the pathogenesis of diabetic cardiomyopathy, evidence is emerging that cardiac dysfunction in the diabetic heart is related to perturbations in myocardial metabolism caused primarily or secondarily by insulin deficiency or resistance. In uncontrolled diabetes, the myocardial extraction and utilization of fat is markedly increased such that the diabetic heart relies almost exclusively on mitochondrial fatty acid oxidation (FAO) for its ATP requirements. Recent studies have defined an important role for the lipid-activated transcription factor, the peroxisome proliferator-activated receptor alpha (PPARalpha), in the control of cardiac fatty acid utilization pathways. Our preliminary data indicates that the activation of cardiac fatty acid utilization in the diabetic heart is mediated by the PPARalpha gene regulatory pathway. Our preliminary data indicates that the activation of cardiac fatty acid utilization in the diabetic heart is mediated by the PPARalpha gene regulatory pathway. This proposal is designed to test the hypothesis that lipid metabolic alterations secondary to increased activity of PPARalpha lead to pathologic remodeling in the diabetic heart. Such pathologic remodeling could occur due top increased oxygen consumption or through toxic lipid intermediates generated by peroxisomal or mitochondrial pathways. This hypothesis will be tested by the phenotypic characterization of mice with cardiac-specific over- expression of PPARalpha (MHC-PPAR mice). First, the lipid metabolic and cardiac functional phenotypic of multiple independent lines of MHC-PPARalpha transgenic mice will be evaluated and compared with that of mice rendered diabetic via administration of streptozotocin. Second, the role of PPARalpha in the expression and severity of diabetic cardiomyopathy will be determined by altering its activity via genetically engineered loss-of-function (PPARalpha null mice) and gain- of-function (MHC-PPAR mice) in the context of three different murine models of diabetes. Lastly, the role of altered peroxisomal function in MHC-PPAR mice compared to diabetic mice. The long term goal of this project is to delineate the precise molecular and metabolic bases for diabetic cardiomyopathy including identification of specific lipid mediators of cardiac dysfunction. This work should lead to the development of novel therapeutic strategies aimed at modulating cardiac lipid metabolism to reduced the cardiovascular morbidity and morality in diabetic patients.

human therapy evaluation; hypopituitarism; somatotropin; hormone therapy; hypoadrenalism; brain injury; disease /disorder proneness /risk; muscle strength; quality of life; body composition; socioeconomics; neuropsychology; patient oriented research; clinical research; human subject; neuropsychological tests;

experimental designs; heart; diabetes mellitus; interdisciplinary collaboration;

heart; diabetes mellitus; experimental designs; interdisciplinary collaboration;

interdisciplinary collaboration; heart; diabetes mellitus;

heart; diabetes mellitus; experimental designs; interdisciplinary collaboration; technology /technique development;

heart; diabetes mellitus; experimental designs; interdisciplinary collaboration;

[unreadable] DESCRIPTION (provided by applicant): [unreadable] The overall goal of this proposal is to establish a planning process to develop novel interdisciplinary strategies to rapidly translate discovery to reduce the burden of diabetic cardiovascular disease, a serious and common medical problem with profound implications for world health. This planning proposal will involve the collective efforts of an interdisciplinary team spanning across 13 departments and 4 colleges at Washington University in St. Louis in partnership with a national population outcomes group. The planning process will be conducted within the framework of the following three elements: Fundamental Discovery to address the pathobiology of diabetic heart and vascular disease; a Technological Platform to support interdisciplinary research activities; and Translational Studies and Clinical Implementation to move discovery to new paradigms in patient care. The process will begin as an open, university-wide dialogue to address existing barriers to transforming "multidisciplinary" research to true "interdisciplinary" units. Planning groups linked to Fundamental Discovery and Translation Pathways will be formed. The Planning groups will come together to develop research strategies that integrate disciplines, establish novel cross-disciplinary training approaches, and evaluate and validate candidate biomarkers aimed at the early detection and risk stratification of myocardial and vascular disease in the diabetic. Rigorous phenotyping strategies will be devised for studies in small groups of humans to assess the potential utility of molecular, biochemical, and imaging markers to detect disease. Plans to validate biomarkers will be developed with population outcomes collaborators. An important long-term goal of the planning process is to outline strategies aimed at the development of a panel of biomarkers that will revolutionize the way we care for diabetic patients through early detection of myocardial and vascular disease, risk stratification, and therapeutic decision-making. It is envisioned that principles established by the fundamental paradigm shift developed through our efforts will be applicable to other complex disease states in the future. [unreadable] [unreadable]

human therapy evaluation; hypopituitarism; somatotropin; subarachnoid hemorrhage; cerebral aneurysm; disease /disorder proneness /risk; hormone therapy; brain injury; hypoadrenalism; quality of life; neuropsychology; patient oriented research; neuropsychological tests; human subject; clinical research;

DESCRIPTION (provided by applicant): To decrease the morbidity and mortality of cardiovascular disease in patients with diabetes, we propose to establish a Specialized Center for Clinically-Oriented Research (SCCOR) in Cardiac Function and Disease at Washington University. The central unifying theme of the proposed SCCOR is to eliminate the excess burden of myocardial disease in people with diabetes. The program is designed to test the hypothesis that derangements in myocardial fatty acid metabolism leads to cardiac dysfunction and increased susceptibility to ischemic insult. We propose that metabolic (diabetes), genetic, racial, and clinical determinants influence the outcome of patients at risk for an acute coronary ischemic event. This SCCOR proposal envisions a multidisciplinary approach involving five Research Projects and four Core Units. The focus of this highly interactive proposal will span from fundamental studies of mouse models of the diabetic heart to outcomes studies in humans. Our approach will combine molecular genetics, development and characterization of genetically modified mice, mechanism-based cardiovascular phenotyping in humans, and population outcomes research. We will examine the role of alterations in myocardial metabolism related to the PPARalpha gene regulatory pathway in response to ischemic insult in mouse models of the diabetic heart (Project 1); define the role of myocardial lipotoxicity in the development of diabetic cardiac dysfunction in mice and humans (Project 2); delineate the contribution of increased myocardial fatty acid metabolism to pathologic ventricular remodeling in patients with diabetes mellitus following coronary ischemia/reperfusion or myocardial infarction using innovative metabolic imaging approaches (Project 3); investigate the basis of racial disparities, lipid metabolic derangements, and genetic factors in outcomes following myocardial infarction (Project 4); and define pharmacogenetic predictors of outcome in diabetic and non-diabetic patients following acute coronary syndrome (Project 5). The long-term objective of this SCCOR is to develop a rigorously defined risk-stratifying panel of imaging, biochemical, genetic, and clinical determinants comprising a phenotypic profile of the patient with diabetes at risk for a cardiovascular event. INDIVIDUAL PROJECTS AND CORE UNITS: PROJECT 1: Altered PPARa Signaling in the Ischemic Diabetic Heart (Kelly, D.) DESCRIPTION (provided by applicant): This SCCOR project will focus on abnormalities of myocardial lipid metabolism in the diabetic patient. Chronically increased rates of fatty acid utilization in the diabetic heart predispose to cardiotoxic effects related to increased oxygen consumption and accumulation of intracellular lipids ("lipotoxicity"). In the setting of myocardial ischemia, high rates of mitochondrial fatty acid oxidation (FAO) may lead to increased myocyte injury and death. Recently, we have found that the nuclear receptor, peroxisome proliferators activated receptor a or PPARa drives increased fatty utilization in the diabetic heart. This project will test the hypothesis that metabolic derangements due to chronic activation of the cardiac PPARa pathway are a major determinant of heart failure and death in diabetics following acute coronary isehemic insult. We have developed mouse models to reproduce the lipid metabolic derangements of the diabetic heart. Transgenic mice with cardiac-specific overexpression of PPARa (MHC-PPAR) exhibits a metabolic phenotype remarkedly similar to the diabetic heart. A second model involves cardiac-specific overexpression of lipoprotein lipase to increase delivery of fatty acids to the heart. We will study the response of the mouse models to ischemic insult. First, the mouse models will be used to evaluate the metabolic and functional response to myocardial infarction and ischemia/reperfusion. Second, we will evaluate the contributory role of cardiac lipotoxicity in the diabetic cardiomyopathic phenotype by evaluating the influence of dietary fat content and the effects of increased or decreased delivery of lipoprotein-derived fatty acid using LPL transgenics and "knockouts", respectively. Third, we will evaluate the influence of pharmacologic agents targeted at the PPAR pathway and its target genes. Lastly, we will perform functional studies of the effects of common single nucleotide polymorphisms (SNPs) within genes of the PPARa complex to compliment the results of population studies planned in Projects 4 and 5. The long-term goal of this project, in collaboration with Projects 2-5, is to identify novel lipid biochemical, metabolic imaging, and genetic determinants predictive of outcome in an individual diabetic patient at risk for acute cornary ischemic insult. (End of Abstract)

[unreadable] DESCRIPTION (provided by applicant): [unreadable] The overall goal of this proposal is to establish a planning process to develop novel interdisciplinary strategies to rapidly translate discovery to reduce the burden of diabetic cardiovascular disease, a serious and common medical problem with profound implications for world health. This planning proposal will involve the collective efforts of an interdisciplinary team spanning across 13 departments and 4 colleges at Washington University in St. Louis in partnership with a national population outcomes group. The planning process will be conducted within the framework of the following three elements: Fundamental Discovery to address the pathobiology of diabetic heart and vascular disease; a Technological Platform to support interdisciplinary research activities; and Translational Studies and Clinical Implementation to move discovery to new paradigms in patient care. The process will begin as an open, university-wide dialogue to address existing barriers to transforming "multidisciplinary" research to true "interdisciplinary" units. Planning groups linked to Fundamental Discovery and Translation Pathways will be formed. The Planning groups will come together to develop research strategies that integrate disciplines, establish novel cross-disciplinary training approaches, and evaluate and validate candidate biomarkers aimed at the early detection and risk stratification of myocardial and vascular disease in the diabetic. Rigorous phenotyping strategies will be devised for studies in small groups of humans to assess the potential utility of molecular, biochemical, and imaging markers to detect disease. Plans to validate biomarkers will be developed with population outcomes collaborators. An important long-term goal of the planning process is to outline strategies aimed at the development of a panel of biomarkers that will revolutionize the way we care for diabetic patients through early detection of myocardial and vascular disease, risk stratification, and therapeutic decision-making. It is envisioned that principles established by the fundamental paradigm shift developed through our efforts will be applicable to other complex disease states in the future. [unreadable] [unreadable]

disease /disorder proneness /risk; pathologic process

disease /disorder proneness /risk; pathologic process

disease /disorder proneness /risk; pathologic process

disease /disorder proneness /risk; pathologic process

This SCCOR project will focus on abnormalities of myocardial lipid metabolism in the diabetic patient. Chronically increased rates of fatty acid utilization in the diabetic heart predispose to cardiotoxic effects related to increased oxygen consumption and accumulation of intracellular fipids ("lipotoxicity"). In the setting of myocardial ischemia, high rates of mitochondrial fatty acid oxidation (FAO) may lead to increased myocyte injury and death. Recently, we have found that the nuclear receptor, peroxisome proliferator-activated receptor alpha or PPARalpha, drives increased fatty utilization in the diabetic heart. This project will test the hypothesis that metabolic derangements due to chronic activation of the cardiac PPARalpha pathway are a major determinant of heart failure and death in diabetics following acute coronary isehemia insult. We have developed mouse models to reproduce the lipid metabolic derangements of the diabetic heart. Transgenic mice with cardiac-specific overexpression of PPARalpha (MHC-PPAR) exhibits a metabolic phenotype remarkedly similar to the diabetic heart. A second model involves cardiac-specific overexpression of lipoprotein lipase to increase delivery of fatty acids to the heart. We will study the response of the mouse models to ischemic insult. First, the mouse models will be used to evaluate the metabolic and functional response to myocardial infarction and ischemia/reperfusion. Second, we will evaluate the contributory role of cardiac lipotoxicity in the diabetic cardiomyopathic phenotype by evaluating the influence of dietary fat content and the effects of increased or decreased delivery of lipoprotein-derived fatty acid using LPL transgenics and "knockouts", respectively. Third, we will evaluate the influence of pharmacologic agents targeted at the PPAR pathway and its target genes. Lastly, we will perform fimctional studies of the effects of common single nucleotide polymorphisms (SNPs) within genes of the PPARalpha complex to compliment the results of population studies planned in Projects 4 and 5. The long-term goal of this project, in collaboration with Projects 2-5, is to identify novel lipid biochemical, metabolic imaging, and genetic determinants predictive of outcome in an individual diabetic patient at risk for acute cornary ischemic insult.

This workshop at Purdue University draws together a global group of interdisciplinary scholars who, together with Purdue faculty, offer analyses of a series of pressing global challenges. The workshop focuses on finding new perspectives on seemingly "intractable" global policy problems by examining a cutting edge area of research in the social sciences: the role of social norms, considered as a kind of informal institution, in shaping policy design, adoption, and implementation. The workshop aims to apply these theoretical insights on multipe difficult problems that have faced policy makers internationally. The workshop involves a series of substantive issue panels focusing on the actual and/or potential role of informal institutions in offering solutions. It also includes an intergrative, cross-issue panel drawing out broader lessons about informal institutions as they relate to multiple diverse issues. The presentations and broader conclusions of this work will be disseminated in both scholarly and non-scholarly outlets.

With regards to intellectual merit, informal institutions (or norms) are increasingly recognized as vital determinants of human behavior in multiple contexts and disciplines. Where simpler models of rationality once dominated our understanding of human behavior, more complicated models emphasizing the influence of norms are now recognized as playing a critical, but poorly understood role. While much progress has been made in the past decade in studying these informal institutions in multiple disciplines and as they operate in many different contexts, the intellectual challenges of understanding the impact of these norms remain daunting and the integration of this work remains limited. By convening this interdisciplinary, cross-national group of scholars, the organizers aim to contribute to the understanding of solutions to policy challenges as well as to advance theoretical understanding of the ways that informal institutions and social norms affect global problems.

With regards to broader impacts, the workshop will offer new solutions to serious problems, promote the professional and intellectual development of graduate students working on vital scientific questions that affect serious policy problems, and build international, interdisciplinary networks and partnerships that include scholars and students from underrepresented gender, racial, and geographic groups.

During the course of this project Indiana University (IU)-Northwest will provide scholarship and educational support to 26 academically talented students, financially disadvantaged students, pursuing Bachelor of Science degrees in the disciplines of biology, chemistry, computer information systems, geosciences, and mathematics. Students will be drawn from two groups: 1) two-thirds of the scholars will be freshmen recruits from local high schools with high average SAT and ACT scores who are in a Dual Credit/Early College attendance program; and 2) the balance of the scholars will be transfer students from area community colleges in Indiana and neighboring Illinois. In addition to receiving financial support, students will also have internship opportunities with area industrial, government, and non-profit entities, thus helping to answer local and regional STEM workforce needs. The university administration and local and regional business communities have indicated strong support for sustaining the program beyond the term of this grant funding.

Beyond the provision of financial assistance, the project's principal activities will be guided by four objectives: effective recruitment; support for high academic performance; persistence and retention in the major; and excellent preparation for jobs or graduate school. To this end the project will initiate Seminars toward Effective Placements (STEM STEPS) to help students advance through their selected program at the highest possible academic level using a combination of cohort style activities, remediation as appropriate, workshops, and peer-led instructional and leadership opportunities. Each year specific classes will be designated as cohort classes (though open to all students) to provide a common educational experience for the scholarship recipients and to build upon the camaraderie established during STEM STEPS. Each cohort will become the peer leaders for the next cohort group, thus providing a continuing cycle of academic and social support during each year of the program. Evaluation of the project will be conducted by an experienced evaluator with a first-hand understanding of academic programs on a regional urban commuter campus. Direct measures of student outcomes will include student performance in STEM courses on various assignments and tests that are related to student learning outcomes as stated on the course syllabi. Evaluation will also examine the results of involvement in internship and faculty mentoring and research activities. The scholarship recipients will form an experimental group whose outcomes will be compared to those of a matched group of students who do not participate in the activities provided through this project. Indirect measures of program success will include anonymous surveys, interviews, and focus groups with students and other stakeholders. Findings will help to advance understanding of what program activities work and the conditions and circumstances under which they work, to improve the success and persistence in STEM learning of the scholarship recipients. As appropriate, student research results from faculty mentored research will be presented at discipline specific scientific meetings at the local, regional, or national level. In addition findings of the program evaluation will be disseminated to the larger scientific community through broadly attended venues such as the annual meeting of the American Association for the Advancement of Science.

SUMMARY Current therapies for heart failure (HF) are largely directed at maladaptive extra-cardiac neurohormonal circuits in a ?one size fits all? approach. There is a significant unmet need for mechanism-based therapies directly targeting the heart during early stages of HF. Increasing evidence has shown that during the development of heart failure, mitochondrial generation of ATP becomes dysregulated. A well-established metabolic signature of the failing heart is a shift from using fatty acids as the chief fuel source of the normal heart, to other fuels such as glucose. This fuel shift occurs early in the development of cardiac hypertrophy and failure. However, the potential linkage of this cardiac fuel switch to the progressive diminution in mitochondrial respiratory function and ATP producing capacity during the development of HF has not been established beyond a mere association. During the current funding period, we have made a series of discoveries that support the premise that disturbances in cardiac fatty acid oxidation (FAO) contribute to mitochondrial energetic dysfunction and the development of HF including: 1) identification of distinct ?bottlenecks? in the terminal steps of the FAO pathway setting the stage for depletion of key cofactors such as Coenzyme A (CoA) and diversion of reducing equivalents away from complex I of the electron transport chain; 2) the ketone body, 3-hydroxybutryate (3OHB), an efficient cardiac fuel that bypasses long-chain FAO, reduces cardiac remodeling and ventricular dysfunction in small and large animal models of HF; and 3) increasing cardiac mitochondrial oxidative capacity, including FAO flux, by cardiac-specific deletion of the gene encoding RIP140 (Nrip1) prevents cardiac hypertrophic growth and reduces cardiac remodeling and dysfunction caused by pressure overload in mice. These findings have led to the central hypotheses of this multi-PI R01 renewal proposal: Downregulation of FAO in the hypertrophied heart results in bottlenecking within the ?-oxidation spiral leading to reduced capacity for mitochondrial ATP production and; reduced FAO flux sets the stage for utilization of carbon sources from glucose and other sources in anabolic pathways necessary for cardiac hypertrophic growth. These hypotheses will be tested by two aims. In Aim 1, we will conduct in-depth assessment of the cardiac functional, mitochondrial, proteomic and genomic response of wild-type, csRIP140-/- (high FAO), and csPPAR?-/- (low FAO) mice during development of HF in mice. Aim 2 is designed to determine the mechanisms whereby RIP140 deficiency defends against pathological cardiac hypertrophic growth. The long-term objectives of the proposed work are to define the mechanistic events leading to mitochondrial energetic collapse in the failing heart and to identify nodal regulatory points that could serve as candidate therapeutic strategies aimed at re-balancing fuel utilization and enhancing mitochondrial ATP-producing capacity aimed at the early stages of heart failure.

SUMMARY The incidence of heart failure (HF), a global health threat, is growing. Current therapies for HF are largely directed at maladaptive extra-cardiac neurohormonal circuits in a ?one size fits all? approach. Evidence has emerged that myocardial fuel and energy metabolic disturbances contribute to the early stages of HF leading to a vicious cycle of energy starvation and contractile dysfunction. We have conducted myocardial metabolomic and proteomic profiling in well-defined mouse models of early stage HF and in the end-stage failing human heart. The results of these profiling studies have identified protein and metabolite signatures in HF that are indicative of bottlenecks in cardiac fatty acid oxidation (FAO), the chief source of acetyl-CoA for the TCA cycle along with evidence for increased ketone body oxidation in the hypertrophied and failing heart. More recently, we have found that increasing delivery of the ketone body, 3-hydroxybutyrate (3OHB) to heart, retards the development of HF in mice and in a canine tachypacing model. However, the biological mechanisms accounting for the cardioprotective effect of 3OHB are unknown. For example, are these beneficial effects cardiac autonomous? Is 3OHB providing a fuel or does it act via other mechanisms? This MPI proposal is designed to test the hypothesis that increasing myocardial ketones reduces pathological cardiac remodeling by providing a more readily oxidizable fuel for mitochondrial ATP production. To address this hypothesis: 1) we will probe the cardiac-specific beneficial actions of 3OHB during the development of HF. The efficacy of a panel of orally administered ketone esters on cardiac function and pathological remodeling will be assessed in wild-type and cardiac-specific Bdh1-deficient mice (unable to oxidize 3OHB in heart) during development of HF. In addition, the cell-autonomous impact of 3OHB on contractility and calcium transients will be assessed in cardiac myocytes isolated from humans with HF; 2) we will investigate the role of 3OHB as a myocardial fuel in its beneficial actions on pathologic cardiac remodeling by assessing the efficacy of R-3OHB (oxidized) vs S-3OHB (unoxidized) ketone esters on the development of HF in mice; and 3) we will develop and validate strategies to increase myocardial 3OHB delivery as a therapeutic strategy for HF in a large animal model by assessing the effects of R-3OHB and S-3OHB on cardiac hemodynamics and substrate metabolism in a canine tachypacing model of progressive HF. Lastly, the potential of 3OHB as a therapeutic agent will be explored by comparing the impact of administration before and after the onset of HF as well as testing oral 3OHB esters. The planned studies will provide important new insight into the mechanisms whereby 3OHB ameliorates HF and will provide in-depth pre- clinical assessment of increasing delivery of ketone bodies to heart as a novel therapeutic.