THE ROLE OF GRP AND GRPERGIC CIRCUITRY IN FEAR MEMORY

Title: THE ROLE OF GRP AND GRPERGIC CIRCUITRY IN FEAR MEMORY

Fiscal Year: 2009

Project Number: 1R01MH080328-01A2

Serial Number: 80328

Funding: NIMH

Principal Investigator: SHUMYATSKY, GLEB P

Abstract: DESCRIPTION (provided by applicant): Sensory information reaching the brain is processed by region-restricted neural networks leading to learning and adaptation to a constantly changing environment. Recent studies have elucidated some of the general molecular mechanisms underlying learning and memory; however, it becomes increasingly clear that there are gene networks that uniquely position key neural circuits to control specialized behaviors. The long-term goal of this work is to systematically characterize how gene regulatory networks control neural circuits dedicated to learned fear. More specifically, this proposal is focused on a gene, gastrin-releasing peptide (GRP), that this laboratory has previously identified as expressed in the neural circuits specifically involved in processing fear- related conditioned stimulus (CS) information. Recently published work from this laboratory suggests that GRP and neuronal circuits that express GRP have an important role in fear memory. The hypothesis will be tested that GRPergic neural circuitry is involved in unimodal and multimodal types of fear memory. A multidisciplinary approach that combines mammalian genetics, behavior and electrophysiology will be used to address the following Specific Aims. Aim 1 will ask whether GRP in the amygdala regulates fear memory. GRP knockout mice will be analyzed in amygdala synaptic plasticity and memory in fear conditioning. The anatomic site of GRP function will be examined by GRP administration in the amygdala of the GRP knockout mice. Aim 2 will ask whether selective elimination of the GRPergic neurons in two different areas of the amygdala differentially affects unimodal and multimodal memories of fear. Aim 3 will ask whether there are two GRP expressing neural microcircuits in the amygdala processing differentially unimodal and multimodal fear memories. GRP and GRPR can be used to develop drugs, targeting neural circuits dedicated to fear and anxiety, and thus more effective interventions can be envisioned that lack side effects. PUBLIC HEALTH RELEVANCE Gastrin-releasing peptide (GRP) may be used to design drugs for the selective regulation of learned fear- related behaviors. Moreover, GRP knockout mice can be used to study how memory of fear is involved in borderline personality disorder, PTSD and generalized anxiety disorder.

BEHAVIORAL AND GENETIC BIOMARKER DEVELOPMENT FOR AUTISM AND RELATED DISORDERS

Fiscal Year: 2009

Project Number: 1RC1MH088288-01

Serial Number: 88288

Funding: NIMH

Principal Investigator: BRZUSTOWICZ, LINDA M

Abstract: DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (03) Biomarker Discovery and Validation and specific Challenge Topic 03-MH-101*: Biomarkers in Mental Disorders. The overall goal of this proposal is to advance the development of behavioral and genetic biomarkers for autism and related disorders. While it is clear that autism has a strong inherited genetic component, very large scale genetic studies that have relied only on a general diagnosis of autism (spectrum) disorders (or other information only on affected individuals) have had limited success in identifying risk alleles, leaving a critical issue for the field. Clearly, alternative genetic study designs are needed to complement existing studies. Behavioral biomarkers, especially language ability, have been used with some success to increase power in gene mapping, but to date studies have focused on detailed behavioral assessments only of subjects with autism and not their family members, despite an extensive literature defining increased rates of related phenotypes in family members. This critical gap will be filled by our project. We will use our existing family set with an extensive existing database of clinical and genetic data from all family members, where each family contains at least one proband with autism and at least one proband with a language deficit, to define biomarkers for risk. For Aim 1, Behavioral Biomarker Development, we will develop a set of behavioral biomarkers of genetic risk for autism and related disorders. We will analyze our extensive behavioral testing database to determine which measures have the strongest genetic effects and further examine latent class structures for both data reduction and to reduce measurement error. We will conduct follow-up assessments on a subset of study participants to determine longitudinal stability of selected biomarkers. For Aim 2, Behavioral Biomarker Validation, we will validate inherited components of the behavioral biomarkers through the use of genome-wide analysis. We will use analysis of quantitative and dichotomous behavioral biomarker data using a quasi-Bayesian posterior probability method to elucidate the genetic architecture of risk, providing evidence of the nature of the inherited genetic component. For Aim 3, Genetic Biomarker Identification, we will identify specific DNA variations associated with genetic risk for autism and related disorders. We will test both common and rare variants, SNPs and CNVs, from candidate genes within the regions identified in Aim 2 and evaluate additional variants from the literature as potential factors modulating a network of risk-determining genes. Overall, we plan to combine analysis of behavioral and genetic biomarkers to develop more accurate models for the prediction of risk for autism spectrum disorders. Our existing detailed clinical and behavioral information, as well as plans for follow-up assessments, will also provide important preliminary data for future comparative effectiveness studies on elements of clinical course and treatment response related to specific biomarkers. It is hoped that these studies will provide substantive insights into the causes of, and effective treatments for, autism. Autism is a serious and debilitating disorder. While there is strong evidence supporting a significant genetic component to the disorder, the identification of specific susceptibility genes has been difficult. Identification of susceptibility genes through the approaches proposed could provide important insights into biological basis of this illness, which could result in the development of novel treatments.

EPIGENETIC REGULATION OF MICRORNAS IN NEUROGENESIS

Fiscal Year: 2009

Project Number: 1R21MH085088-01A1

Serial Number: 85088

Funding: NIMH

Principal Investigator: HART, RONALD P

Abstract: DESCRIPTION (provided by applicant): Epigenetic regulation of microRNAs in neurogenesis Histone deacetylase (HDAC) inhibitors such as valproate (VPA) are commonly used to treat epilepsy. One me- chanism of their efficacy may result from epigenetic effects on differentiation of neural precursors. While HDAC inhibitors are likely to affect mRNA-encoding genes, we believe that the rapid regulation of microRNAs by VPA treatment suggests a novel complementary mechanism. Hypothesis: Acetylated histones allow expression of microRNAs that enhance or support neurogenesis from neural precursors. Therefore, adding an HDAC inhibitor allows the accumulation of acetyl marks on microRNA-encoding sites on the chromatin, in turn enhancing expression of the marked microRNAs, which contribute to neural differentiation. We will test the hypothesis by using 'deep sequencing' to identify the full complement of VPA-regulated microRNAs, including hundreds of novel microRNAs that we recently identified in neural differentiation. Finally, we will build on our preliminary studies with microRNA inhibitors and screening techniques to evaluate candidate microRNAs for their requirement for VPA-induced neuronal differentiation. Results from these three aims will identify both known and putatively novel microRNAs that are regulated after HDAC inhibition, the acetylation status of histones located near microRNA promoter regions, and whether the regulated microRNAs contribute to neuronal differentiation. Understanding the regulatory networks required for neurogenesis is important for determining how differentiation of neural precursors could be impaired in conditions such as dementia, Parkinson's, or stroke. Alternatively, the use of HDAC inhibitors in conditions such as epilepsy, bipolar disorder, or migraines may have unexpected effects on neurogenesis from adult precursors, and this potential benefit should be understood. PUBLIC HEALTH RELEVANCE: Epigenetic regulation of microRNAs in neurogenesis. Epilepsy drugs such as valproate (VPA) are known to inhibit histone deacetylases, which control gene expression epigenetically. Since VPA also increases the production of neurons from adult stem cells or other precursors, we believe that VPA may regulate microRNA genes, in turn affecting decisions controlling cell differentiation. We will use 'deep sequencing' of microRNAs and gene regulatory regions to identify novel neurogenesis mechanisms.

EXPERIMENTS & COMPUTATIONS TO FIND AGGREGATION-PRONE ENSEMBLES OF ALPHA-SYNUCLEIN

Fiscal Year: 2009

Project Number: 1R01GM087012-01A1

Serial Number: 87012

Funding: NIGMS

Principal Investigator: BAUM, JEAN S ;LEVY, RONALD ;

Abstract: DESCRIPTION (provided by applicant): a-synuclein (aSyn) is an intrinsically disordered protein that appears in aggregated form in the brains of patients with Parkinson's disease. The conversion of monomer to aggregate is complex. Aggregation rates of aSyn are very sensitive to changes in amino acid sequence and environmental conditions. Understanding aSyn aggregation requires characterizing the ensemble of conformations adopted by the monomer and correlating them to aggregation behavior. Though many hypotheses have been proposed to relate aSyn's aggregation behavior to its interconverting conformational ensembles, a consistent molecular description of the aSyn conformational ensembles and their relationship to aggregation remains elusive. This proposal integrates NMR and computational approaches to characterize and explicitly visualize the intrinsically disordered conformational ensembles of aSyn and the early stages of aggregation under different sequence and environmental conditions. The goal is to identify the elements of transient 2o and/or 3o structure that are key for initiation of aggregation and determine their stabilizing driving forces. Identifying the structural basis of aSyn monomer aggregation propensity may be critical for developing inhibitors for the aggregation steps that precede the toxic aggregation cascade. Once developed, this integrated approach can be applied to other important biological functions or diseases involving intrinsically disordered proteins. PUBLIC HEALTH RELEVANCE: Parkinson's disease is the second most prevalent of the late onset neurodegenerative diseases. a-synuclein, an extremely important protein involved in the etiology of Parkinson's disease will be modeled at the molecular level by integrating NMR and computational approaches. Understanding the role of the monomeric conformational ensembles of a-synuclein may be critical for developing inhibition strategies against amyloid formation.

REGULATION OF APICAL SPECIFIC ENDOCYTOSIS IN THE C. ELEGANS INTESTINE

Fiscal Year: 2009

Project Number: 1R21DK082854-01

Serial Number: 82854

Funding: NIDDK

Principal Investigator: GRANT, BARTH

Abstract: DESCRIPTION (provided by applicant): Our goal in this proposal is to identify and understand the cellular components that internalize and transport endocytic cargo from the apical plasma membrane to the lysosomes of intestinal epithelial cells (enterocytes). Many epithelial cell surface receptors, ion channels, and other integral membrane proteins implicated in human disease are specifically localized to the apical membrane. Their steady-state abundance on the apical surface is largely controlled by their relative rates of insertion and removal from the plasma membrane by secretion and endocytosis. To gain new insight into the mechanisms that drive this pathway, we propose to pioneer the use of the microscopic nematode worm C. elegans for these studies. We will take advantage of the unique experimental features of this system, creating a new, highly simplified, and genetically manipulatable paradigm for this research area. Chief among the features that have made C. elegans a leading model organism in nearly all areas of modern biological research are its highly advanced genetics, including extremely facile gene knockdown, knockout, and transgenic technology, coupled with a transparent body that allows visualization of fluorescently tagged molecules in the physiologically relevant context of the intact living organism. Our preliminary studies have already uncovered a novel regulatory pathway controlling apical endocytosis through Rac1/CED-10. Further analysis will dissect the mechanisms by which this pathway controls apical endocytosis. In addition, we plan to apply the unique genetic tools available in C. elegans to identify additional regulators of apical endocytosis conserved with mammals. As the simplest animal model possessing a true intestinal epithelium, C. elegans has the potential to answer key questions regarding the mechanism of apical intestinal function, and can provide a key framework that catalyzes future investigation into the highly related pathways in mammalian intestinal epithelia. PUBLIC HEALTH RELEVANCE. Our goal in this proposal is to identify and understand the cellular components that internalize and transport macromolecules from the apical plasma membrane to the lysosomes of intestinal epithelial cells (enterocytes). Many important proteins implicated in human disease are specifically taken up from the apical membrane by endocytosis, which is specialized in enterocyte cells, and displays important differences from related processes in other types of cells. Thus this research will be relevant to nutrient uptake, fluid homeostasis and dysentery, and fetal antibody transport.

PHENOTYPIC MARKERS FOR SMOKING CESSATION: IMPULSIVE CHOICE AND IMPULSIVE ACTION

Fiscal Year: 2009

Project Number: 1RC1DA028129-01

Serial Number: 28129

Funding: NIDA

Principal Investigator: MCCARTHY, DANIELLE ERIN

Abstract: DESCRIPTION (provided by applicant): Phenotypic Markers for Smoking Cessation: Impulsive Choice and Impulsive Action This application addresses broad Challenge Area (01): Behavior, Behavioral Change, and Prevention and specific high-priority Challenge Topic 01-AA-101 Identifying Phenotypic Markers for Positive Behavior Change. The proposed project will gather new information about temporal growth patterns in two phenotypic markers of behavior change among smokers engaged in an attempt to quit smoking with individual smoking cessation counseling and nicotine lozenge treatment. The project will also generate real time data about the associations between alcohol use and measures of impulsive choice and action in individuals attempting to stop smoking. Delay discounting is a devaluing of delayed rewards that has been documented in smokers, particularly during acute periods of withdrawal. Delay discounting has important but previously unexplored implications for momentary decisions that smokers attempting to quit must make regarding the value of abstaining now, despite withdrawal discomfort and powerful cravings, to reap health benefits years later. The proposed project will examine day-to-day fluctuations in delay discounting assessed daily by palmtop computer in the 7 days preceding a quit attempt and the first 21 days of a quit attempt and will explore relations between delay discounting and subsequent abstinence. Subjects will also complete a standard, laboratory-based delay discounting task three times in the study (one week pre-quit, on the target quit day, and 3 weeks post-quit) which will be assessed as predictors of later abstinence. Variables that may account for changes in delay discounting over time, such as alcohol use, affective states, access to cigarettes, or perceived certainty of health benefits of quitting, will be examined as well. In addition to the delay discounting measure of impulsive choice, the proposed project will also administer a task assessing impulsive action, Go-no go tasks require that individuals be able to inhibit prepotent responses, an ability with clear relevance for smoking cessation. Subjects are instructed to tap a key when a go stimulus appears and to withhold the response when the signal is not present. To date, real-time data regarding impulsive action have not been collected during the course of a quit attempt. This project will generate new information about changes in impulsive action and factors associated with such changes (e.g., alcohol use, craving) during an attempt to inhibit over-learned smoking behavior. The within-subjects longitudinal design of the proposed project will generate new knowledge about change in impulsive choice and impulsive action, and the associations between these constructs over time, during an assisted behavior change attempt. The project will also generate new information about relations between real-time behavioral measures of impulsivity and affective states, environmental contexts, behaviors (including alcohol use) assessed in real time, and subsequent abstinence. The proposed research may help identify cognitive markers of successful change or risk that could be targeted in future interventions. The proposed research will combine addiction research with cognitive psychology and behavioral economics to fill an important gap in our understanding about dynamic changes in decisions and behaviors that may influence smoking behavior. Tracking preference for immediate versus delayed rewards and impulsive action during an attempt to quit smoking and may help us identify markers of successful cessation with implications for other substances of abuse, including alcohol.

ROLE OF TUMOR AND STROMAL CELL METABOLISM IN STRESS ADAPTATION AND PROGRESSION

Fiscal Year: 2009

Project Number: 1RC1CA147961-01

Serial Number: 147961

Funding: NCI

Principal Investigator: COLLER, HILARY A;RABINOWITZ, JOSHUA D;WHITE, EILEEN ;

Abstract: DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (15) Translational Science and specific Challenge Topic, 15-CA-112: Cancer Cell Energy Metabolism and Cancer Causation The molecular events that covert normal cells into tumor cells alter cellular metabolism to aerobic glycolysis, which favors macromolecular synthesis at the expense of efficient ATP production. This metabolic switch that facilitates tumor growth remains a fundamental distinction between normal and tumor cells that has yet to be effectively exploited of cancer therapy. Deregulated tumor cell growth and altered metabolism are also a source of metabolic stress. Oxygen, nutrient and factor deprivation caused by growth to high density and insufficient angiogenesis, and lactate production from glycolysis, are common features and sources of stress in the tumor microenvironment. How tumor and associated stromal cells respond to this stress effects tumor progression and treatment response. Possible responses include cell death (apoptosis or necrosis), cell cycle exit (quiescence or senescence) or adaptation and survival (autophagy). The catabolic process of autophagy is a lysosomal degradation pathway induced by metabolic stress that confers stress tolerance by maintaining energy homeostasis through cellular self-consumption and recycling, and by mitigating oxidative damage through the degradation of malfunctioning organelles and proteins. Autophagy may be particularly important to cells in a glycolytic state and under conditions of metabolic stress. Thus, the adaptation of tumor and associated stromal cells to metabolic stress is intimately linked to their inherent metabolic activity, but the role of metabolism in the context of tumor-stromal interaction is not known. Moreover, exactly how autophagy contributes to cellular metabolism and survival in the tumor microenvironment is not clear. We hypothesize that metabolic adaptation in tumor tissue allows prolonged survival to stress permitting tumor relapse and that discerning the underlying mechanisms will provide new approaches to cancer therapy. To test this hypothesis, we plan to define tumor and stromal cell metabolism, the role of catabolism through autophagy, and the mechanism of metabolic stress adaptation. Growth to high density induces fibroblasts to alter metabolism, activate autophagy, exit the cell cycle, and enter quiescence. We discovered that induction of quiescence of stromal fibroblasts dramatically alters gene expression, including activation of Notch signaling that is required for cell cycle reentry. Remarkably, glucose consumption and utilization is high in quiescent fibroblasts, which causes lactate secretion which can alter the microenvironment; but how does it influence tumorigenesis is not known. Analogous to quiescence in fibroblasts, apoptosis-defective tumor cells subjected to metabolic stress activate autophagy, exit the cell cycle and enter a state of prolonged dormancy from which they can reenter the cell cycle when growth conditions are favorable. We discovered that tumor cell dormancy dramatically alters the cellular proteome and gene expression with evidence of compensatory nutrient uptake, Foxo and Notch pathway activation, and induction of uncharacterized mammalian homologues of yeast genes required for quiescence induced by carbon source limitation. These findings suggest that major metabolic reprogramming accompanies the transition from proliferation to dormancy to cell cycle reentry. These striking parallels between quiescence and dormancy suggest that they are governed by common metabolic reprogramming events that may be a fundamental aspect of tumor biology that has yet to be explored. We propose to define the metabolic networks and role of metabolism as cells transition from proliferation to dormancy or quiescence to cell cycle reentry and how this alters tumor-stromal interaction. Preventing tumor cells from successful metabolic adaptation to stress enabling sustained dormancy and recovery may provide a novel approach to cancer therapy. Cancer cells acquire mutations that deregulate cell growth that also alter cellular metabolism. It has recently become apparent that the metabolic reprogramming of tumor cells is necessary to provide the building blocks for the macromolecular synthesis of proteins, lipids and nucleic acids necessary to support tumor cell growth. Importantly, these metabolic alterations distinguish tumor cells from normal cells, providing a potential therapeutic window and novel targets for anti-cancer drug discovery. PUBLIC HEALTH RELEVANCE: Cancer cells acquire mutations that deregulate cell growth that also alter cellular metabolism. It has recently become apparent that the metabolic reprogramming of tumor cells is necessary to provide the building blocks for the macromolecular synthesis of proteins, lipids and nucleic acids necessary to support tumor cell growth. Importantly, these metabolic alterations distinguish tumor cells from normal cells, providing a potential therapeutic window and novel targets for anti-cancer drug discovery.

C. ELEGANS AS A MODEL FOR RICIN INTOXICATION

Fiscal Year: 2009

Project Number: 1R21AI077805-01A1

Serial Number: 77805

Funding: NIAID

Principal Investigator: TUMER, NILGUN E

Abstract: DESCRIPTION (provided by applicant): Ricin is a plant toxin produced by the castor bean (Ricinus communis) that is extremely toxic to mammalian cells and is classified as a select agent by the US Department of Health and Human Services (HHS) and a category B priority pathogen by the Center for Disease Control (CDC). Although there have been attempts to generate an effective vaccine against ricin, no recombinant vaccine has yet been approved for human use and there are no effective treatment measures against ricin exposure. Therefore there is an urgent need for therapeutics to protect ricin-exposed individuals. To gain understanding into the mechanism of ricin induced cell death, we will take advantage of the unique experimental features of C. elegans that have made it a leading model organism in nearly all areas of biological research. Our primary objective in this proposal is to establish C. elegans as a new whole animal model to study ricin intoxication and to translate our findings to mammals. In preliminary studies, we fed C. elegans with either E. coli expressing ricin A-chain (RTA) or recombinant RTA purified from E. coli and demonstrated that RTA is lethal to C. elegans at extremely low doses. In agreement with mammalian data, we showed that ricin uses a clathrin and a raft independent endocytosis pathway to enter the C. elegans intestinal epithelial cells and entry to the early endosome is critical for cytotoxicity. We identified a novel role for the two major phagocytosis/engulfment pathways in mediating ricin sensitivity in the C. elegans intestine. These results provided support for the development of C. elegans as a new multicellular model for molecular genetic dissection of the mechanisms of ricin transport and cell death. We will examine transport of ricin in the worm intestine, a classic polarized epithelium, in the context of a living animal. C. elegans mutants defective in intracellular trafficking pathways will be used to validate C. elegans as a relevant model for mechanistic analysis of ricin uptake and transport and to identify critical steps in ricin transport that lead to cytotoxicity. We will exploit powerful features of C. elegans genetics to isolate new mutants defective in ricin intoxication. The proposed studies will lead to elucidation of the molecular details of ricin transport and will lay the foundation for identification of inhibitors that can prevent ricin induced cell death. PUBLIC HEALTH RELEVANCE: Our primary objective in this application is to establish C. elegans as a new model to study the mechanism of ricin intoxication and to translate our findings to humans. These studies are significant because currently there are no vaccines, antidotes or any other treatment measures against ricin exposure. These studies will lay the foundation for identification of inhibitors that can protect ricin exposed individuals.

PAVLOVIAN INFLUENCES ON REWARD-SEEKING BEHAVIOR

Fiscal Year: 2009

Project Number: 1R03DA026559-01

Serial Number: 26559

Funding: NIDA

Principal Investigator: SHIFLETT, MICHAEL WILLIAM

Abstract: DESCRIPTION (provided by applicant): A prevalent form of relapse in addiction is caused by exposure to cues or contexts formerly paired with drug delivery. The ability of drug-paired conditioned stimuli (CSs) to increase incentive motivation, or 'craving', indicates that the rewarding properties of drug exposure can be associated with CSs predicting their delivery through a Pavlovian incentive learning process. Recent research raises the possibility that, as a consequence of neuroadaptations resulting from repeated drug exposure, the ability of CSs to compel reward-seeking behavior is enhanced among former drug users. To fully explore this hypothesis, a greater understanding is necessary of the motivational processes that underlie Pavlovian influences on reward-seeking behavior generally, as well as the effects of repeated drug exposure on these processes. This proposal examines the effect of drug sensitization on the ability of Pavlovian cues to influence reward-seeking behavior, with the overall goal of characterizing the motivational processes that underlie the enhanced responsiveness to reward-associated CSs in amphetamine-sensitized rats. The proposed experiments make use of Pavlovian-instrumental transfer (PIT) paradigm to assess the role of outcome saliency and general arousal in determining the choice and intensity of instrumental actions during PIT. These experiments are important for understanding the process through which drug exposure alters the influence of reward-predictive cues on reward-seeking behavior, and will serve as the basis for further research into the neural systems and molecular mechanisms underlying Pavlovian incentive processes and their modulation by exposure to drugs of abuse. Furthermore, these studies will be important in devising therapeutic strategies to treat relapse based on an understanding of the relevant factors that underlie Pavlovian influences on reward-seeking behavior. PUBLIC HEALTH RELEVANCE: The proposed research addresses behavioral processes that are central to substance abuse disorders, and thus is highly relevant to public health. Furthermore, the research findings may be useful in devising therapeutic strategies to prevent relapse caused by re-exposure to drug-associated cues.

BIOLOGY OF THE NK CELL CYTOLYTIC ACTIVITY RHYTHM

Fiscal Year: 2009

Project Number: 1R01HL088041-01A1

Serial Number: 88041

Funding: NHLBI

Principal Investigator: SARKAR, DIPAK KUMAR

Abstract: Circadian and daily rhythms regulate many aspects of physiology and behavior. Although a growing number of studies suggest that circadian disruptions may render organisms more susceptible to infection and cancer, the molecular links between the circadian system and the immune system are largely unknown. Natural killer (NK) cell cytolytic activity has been shown to follow a daily rhythm, which is under a circadian control. NK cells participate in the immune response against viral infections and cancer. Employing the rat animal model, this proposal determines the mechanisms by which the circadian rhythm of splenic NK cell cytolytic function is regulated. It tests the hypothesis that splenic NK cells are provided with molecular clocks that orchestrate the rhythmic expression of cytolytic factors that drive the circadian rhythm of NK cell cytolytic function. The hypothalamic suprachaismatic nucleus, which is itsetf, entrained by photic input, coordinates the expression of clock regulatory genes by altering the sympathetic outflow to the splenic NK cells. The proposed research uses magnetic separation techniques to isolate the NK cell population from the spleen to determine the changes in cytotoxic factors, cytokines and clock genes in these cells; employs gene knockdown techniques in NK cells to study the influence Of clock genes on cytolytic function; analyzes the circadian variations in the clearance of NK-sensitive tumor cells; determines the susceptibility of NK cell function to jet lag, a common circadian disruption, and uses pharmacological tools to investigate the signaling mechanism critically involved in resetting the circadian clock in splenic and clonal NK cells. The proposed series of studies should enhance our knowledge of the physiological mechanisms orchestrating NK cell circadian function.

PERSISTENCE AND DESISTENCE IN HEAVY DRINKING AND VIOLENCE

Fiscal Year: 2009

Project Number: 1R01AA016798-01A1

Serial Number: 16798

Funding: NIAAA

Principal Investigator: WHITE, HELENE R.

Abstract: The present application takes advantage of the high quality data that already exist and proposes a studythat can be completed within 2 years (at a cost of $737,367). It will combine data from the youngest PYScohort from ages 7 through 25 with data from the oldest cohort from ages 13 to 25 to examinepersistence and desistance in heavy drinking and violence from childhood into young adulthood among ahigh-risk group of White and African American men. Data from the youngest cohort at age 25 have justrecently been collected and there are no funds available to combine those data with the data from theoldest cohort through age 25 to examine the co-occurrence of heavy drinking and violence from childhoodthrough age 25. Within a 2-year period, we will be able to make a significant contribution to the scholarlyliterature and provide data to be used in the development of targeted pre,vention programs.

NANOSCALE RECEPTOR BLOCKERS FOR INHIBITION OF ATHEROSCLEROSIS

Fiscal Year: 2009

Project Number: 1R21HL093753-01A2

Serial Number: 93753

Funding: NHLBI

Principal Investigator: MOGHE, PRABHAS V

Abstract: DESCRIPTION (provided by applicant): Pathologies in blood vessels arising from the uncontrolled build-up of oxidized lipids contribute to atherosclerosis, a severe cardiovascular disease, which underlies the most common cause of adult death in the U.S. (exceeding one million patients yearly). Few existing therapeutic strategies address the local management of atherogenesis (build-up of oxidized lipids in the blood vessel walls) and related inflammation. The overall goals of this study are to rationally design and characterize nanoscale biomaterials as a novel cell-targeted materials platform for investigating strategies to de-escalate the onset of atherogenesis and reduce accompanying inflammation. The proposed NIH R21 study involves three specific objectives to investigate nanoassembled amphiphilic polymers (NAPs) to maximally inhibit oxidized LDL uptake in human macrophages under physiologic conditions and exhibit potential for specific targeting to inflamed endothelia. Efforts in Aim 1 will involve investigation of innovative designs of nano-assembled amphiphilic polymers (NAP) composition and architecture to promote NAP binding to both SRA-1 and CD36 scavenger receptors on human THP-1 macrophages and thus inhibit uptake of oxidized low-density lipoproteins (oxLDL). New configurations of NAP will tested for improved lipid uptake inhibition in the presence of serum. Studies in Aim 2 will investigate the effect of NAP-scavenger receptor (SR) interactions on the downstream intracellular and cell-secreted intermediates regulating atherogenesis in macrophages, including cytokine secretion; cholesterol ester accumulation; matrix metalloproteinase secretion; and expression analysis of genes involved in pro-atherogenic signaling pathways. Aim 3 is concerned with design and evaluation of the potential of biofunctionalized NAPs to bind to and transport across activated endothelial cell cultures in vitro, thereby creating a simplified in vitro model of the rescue of macrophage cells involved in atherogenesis within the vascular intima. PUBLIC HEALTH RELEVANCE: The excessive uptake of modified forms of LDL in immune blood cells macrophages is one of the hallmarks of fat build-up and vascular disease within blood vessel walls, which can lead to blockage of blood flow, and cause heart disease or stroke. This study will investigate the design of nanoscale assembled polymers with specific architectures, charge displays, and chemistry so as to reduce the uptake of the most damaging forms of lipoproteins within macrophages. The goals of the study are to identify the most effective 'nanolipoblocker' configurations that may prevent atherogenesis by targeting activated blood vessel cells and blocking foam cell formation.

MULTIPLEX NANOCARRIER-BASED HYDROGELS FOR PREVENTION OF VAGINAL HIV TRANSMISSION.

Fiscal Year: 2009

Project Number: 1R01AI084137-01

Serial Number: 84137

Funding: NIAID

Principal Investigator: SINKO, PATRICK J.

Abstract: DESCRIPTION (provided by applicant): With the incidence of HIV infection on the rise, the development of vaccines and topical microbicides has been a major worldwide priority. However, the results of recent trials have been disappointing. As such, the induction of sterilizing immunity and protection against HIV infection continues to be a major public health goal. 'Microbicides', topically applied agents that prevent HIV transmission from person to person, are still believed to hold considerable promise. In fact, it has been suggested that a microbicide with only 60% efficacy could prevent about 1 million HIV infections per year. Given recent clinical failures, there is an urgent need to rethink the concept of microbicides. Therefore, the long-term objective of the proposed research is to design, construct and evaluate a multiplex nanocarrier-based polyethylene glycol (PEG) vaginal hydrogel for preventing HIV transmission. PEG is nontoxic and biocompatible. Hydrogels resemble living tissue due to their high-water content and soft/rubbery characteristics. The hydrogel is a liquid upon instillation allowing for high vaginal dispersion/mucosal coverage where it then undergoes a rapid phase transition to form a visco-elastic gel. The proposed gel must be multifunctional since it has been shown that (1) sexually transmitted and genital infections such as bacterial vaginosis (BV) increase the risk of HIV transmission by weakening mucosal barriers and by stimulating an inflammatory response that may activate or recruit HIV target cells to the portals of viral entry, (2) low vaginal pH (<4.5) is essential for the prevention of vaginal infections but is not sufficient to inhibit vaginal pathogens and to prevent infection, and (3) cell-associated HIV breaches the normal vagina stratified squamous epithelial barrier but with low frequency. The gel matrix will be formed by crosslinking various PEG nanocarriers each of which plays a unique role in the functional properties of the hydrogel (e.g., promoting mucosal adhesion, maintaining mildly acidic pH, releasing microbicide and spermicides, and preventing HIV virion binding). We will design, synthesize, characterize, and evaluate a series of crosslinking nanocarriers that impart a variety of functional properties to the microbicide hydrogel. Aim 1: To construct an effective physical viral barrier using a fast forming, degradable hydrogel with high vaginal dispersion, high mechanical strength, and viscoelastic properties. Aim 2: To create nanocarriers possessing acidifying agents using natural acids and/or the microbicide/spermicide subtilosin. Aim 3: To fabricate polyanionic or RGD nanocarriers to prevent free or cell-associated HIV binding. Aim 4: To evaluate the various crosslinking nanocarriers and hydrogels in cell, tissue and animal models. If successful, the proposed research will result in a novel multifunctional hydrogel technology that possesses the ideal properties of an anti-HIV microbicide: it will be colorless, odorless, inexpensive to manufacture, safe to use more than once a day and for long periods of time, fast-acting, undetectable to either partner, and available in contraceptive and noncontraceptive forms. With the incidence of HIV infection on the rise, the development of vaccines and topical microbicides has been a major worldwide priority but the results of recent trials have been disappointing. 'Microbicides', topically applied agents that prevent HIV transmission from person to person, are still believed to hold considerable promise. The proposed research seeks to design, construct and evaluate an instantly-forming multifunctional vaginal hydrogel to prevent the initial infection and dissemination of HIV through the vaginal mucosa to distant tissues in the body.

PHENOTYPIC MARKERS FOR SMOKING CESSATION: IMPULSIVE CHOICE AND IMPULSIVE ACTION

Fiscal Year: 2009

Project Number: 1RC1DA028129-01

Serial Number: 28129

Funding: NIDA

Principal Investigator: MCCARTHY, DANIELLE ERIN

Abstract: DESCRIPTION (provided by applicant): Phenotypic Markers for Smoking Cessation: Impulsive Choice and Impulsive Action This application addresses broad Challenge Area (01): Behavior, Behavioral Change, and Prevention and specific high-priority Challenge Topic 01-AA-101 Identifying Phenotypic Markers for Positive Behavior Change. The proposed project will gather new information about temporal growth patterns in two phenotypic markers of behavior change among smokers engaged in an attempt to quit smoking with individual smoking cessation counseling and nicotine lozenge treatment. The project will also generate real time data about the associations between alcohol use and measures of impulsive choice and action in individuals attempting to stop smoking. Delay discounting is a devaluing of delayed rewards that has been documented in smokers, particularly during acute periods of withdrawal. Delay discounting has important but previously unexplored implications for momentary decisions that smokers attempting to quit must make regarding the value of abstaining now, despite withdrawal discomfort and powerful cravings, to reap health benefits years later. The proposed project will examine day-to-day fluctuations in delay discounting assessed daily by palmtop computer in the 7 days preceding a quit attempt and the first 21 days of a quit attempt and will explore relations between delay discounting and subsequent abstinence. Subjects will also complete a standard, laboratory-based delay discounting task three times in the study (one week pre-quit, on the target quit day, and 3 weeks post-quit) which will be assessed as predictors of later abstinence. Variables that may account for changes in delay discounting over time, such as alcohol use, affective states, access to cigarettes, or perceived certainty of health benefits of quitting, will be examined as well. In addition to the delay discounting measure of impulsive choice, the proposed project will also administer a task assessing impulsive action, Go-no go tasks require that individuals be able to inhibit prepotent responses, an ability with clear relevance for smoking cessation. Subjects are instructed to tap a key when a go stimulus appears and to withhold the response when the signal is not present. To date, real-time data regarding impulsive action have not been collected during the course of a quit attempt. This project will generate new information about changes in impulsive action and factors associated with such changes (e.g., alcohol use, craving) during an attempt to inhibit over-learned smoking behavior. The within-subjects longitudinal design of the proposed project will generate new knowledge about change in impulsive choice and impulsive action, and the associations between these constructs over time, during an assisted behavior change attempt. The project will also generate new information about relations between real-time behavioral measures of impulsivity and affective states, environmental contexts, behaviors (including alcohol use) assessed in real time, and subsequent abstinence. The proposed research may help identify cognitive markers of successful change or risk that could be targeted in future interventions. The proposed research will combine addiction research with cognitive psychology and behavioral economics to fill an important gap in our understanding about dynamic changes in decisions and behaviors that may influence smoking behavior. Tracking preference for immediate versus delayed rewards and impulsive action during an attempt to quit smoking and may help us identify markers of successful cessation with implications for other substances of abuse, including alcohol.

INTEGRATING QUANTITATIVE HISTOLOGICAL IMAGE AND VASCULAR DENSITY PATTERNS FOR PRO

Fiscal Year: 2009

Project Number: 1R03CA143991-01

Serial Number: 143991

Funding: NCI

Principal Investigator: MADABHUSHI, ANANT

Abstract: DESCRIPTION (provided by applicant): With increasing detection of early CaP with improved diagnostic methodologies, it has become important to predict biologic behaviors and 'aggressivity' to identify patients who might benefit from a 'wait and watch policy' as opposed to those who need more aggressive strategies. Traditionally, T-stage, amount of cancer in the core biopsy, the Gleason grade, and PSA at diagnosis has been used to evaluate the prognosis in localized CaP. While the Gleason score is currently assumed to be the strongest prognostic marker for CaP, there is often considerably high inter-, intra-observer variability associated with Gleason grade determination by pathologists. While some newer markers have recently shown promise, none of these methods have individually proven to be accurate enough to serve routinely as a prognostic marker for CaP. Recently, there has been a call to combine multiple prognostic markers to create an integrated meta-marker, with potentially greater accuracy in predicting CaP recurrence compared to any individual marker. While it is apparent that prognostic information resides in histopathology imagery in terms of the arrangement of nuclei and glands, sophisticated graph, and computerized image analysis algorithms are required to quantitatively model and characterize the architectural appearance of prostate cancer histopathology and thus provide a marker that is accurate and reproducible (unlike Gleason grade). In addition, while tumor micro-vascular density has been correlated to CaP outcome, prognostic information may also potentially reside in the specific spatial architectural arrangement of the micro-vascular network. The objective of the proposed work is to develop an integrated quantitative prognostic marker that combines information based on architectural arrangement of nuclear, glandular, and micro-vasculature network patterns on whole mount histology sections (WMHS) obtained via radical prostatectomy (RP) to predict prostate cancer recurrence. The proposed work comprises a total of 3 specific aims. For this project we will digitize approximately 100 annonymized WMHS obtained via RP that have been matched for Gleason score, stage, PSA, but with different clinical outcomes (half the patients having undergone cancer recurrence and the other not, following RP). Under Aim 1, segmentation algorithms will be developed to automatically identify cancerous nuclei, glands and tumor microvasculature (MV), stained immuno-histochemically via CD31. Under Aim 2 we will apply graph based image analysis algorithms to quantitatively characterize the architectural arrangement of CaP nuclei, glands and the MV network. These graph-based features will be integrated via a computerized machine learning algorithm to yield a numerical image based risk score (IbRiS) reflecting the CaP prognosis (disease recurrence or non-recurrence) of the patient. IbRiS will be evaluated in terms of its ability to distinguish between CaP progressors and non-progressors (matched for stage, Gleason grade, PSA), in a cohort of 50 independent studies (test set) for which survival and outcome data is available. This project will be a collaboration between investigators at Rutgers University (RU) and the University of Pennsylvania (UPENN). Data accrual will be done at UPENN while algorithmic development for computerized image analysis and classification will be carried out at RU.

REGULATION OF APICAL SPECIFIC ENDOCYTOSIS IN THE C. ELEGANS INTESTINE

Fiscal Year: 2009

Project Number: 1R21DK082854-01

Serial Number: 82854

Funding: NIDDK

Principal Investigator: GRANT, BARTH

Abstract: DESCRIPTION (provided by applicant): Our goal in this proposal is to identify and understand the cellular components that internalize and transport endocytic cargo from the apical plasma membrane to the lysosomes of intestinal epithelial cells (enterocytes). Many epithelial cell surface receptors, ion channels, and other integral membrane proteins implicated in human disease are specifically localized to the apical membrane. Their steady-state abundance on the apical surface is largely controlled by their relative rates of insertion and removal from the plasma membrane by secretion and endocytosis. To gain new insight into the mechanisms that drive this pathway, we propose to pioneer the use of the microscopic nematode worm C. elegans for these studies. We will take advantage of the unique experimental features of this system, creating a new, highly simplified, and genetically manipulatable paradigm for this research area. Chief among the features that have made C. elegans a leading model organism in nearly all areas of modern biological research are its highly advanced genetics, including extremely facile gene knockdown, knockout, and transgenic technology, coupled with a transparent body that allows visualization of fluorescently tagged molecules in the physiologically relevant context of the intact living organism. Our preliminary studies have already uncovered a novel regulatory pathway controlling apical endocytosis through Rac1/CED-10. Further analysis will dissect the mechanisms by which this pathway controls apical endocytosis. In addition, we plan to apply the unique genetic tools available in C. elegans to identify additional regulators of apical endocytosis conserved with mammals. As the simplest animal model possessing a true intestinal epithelium, C. elegans has the potential to answer key questions regarding the mechanism of apical intestinal function, and can provide a key framework that catalyzes future investigation into the highly related pathways in mammalian intestinal epithelia. PUBLIC HEALTH RELEVANCE. Our goal in this proposal is to identify and understand the cellular components that internalize and transport macromolecules from the apical plasma membrane to the lysosomes of intestinal epithelial cells (enterocytes). Many important proteins implicated in human disease are specifically taken up from the apical membrane by endocytosis, which is specialized in enterocyte cells, and displays important differences from related processes in other types of cells. Thus this research will be relevant to nutrient uptake, fluid homeostasis and dysentery, and fetal antibody transport.

BIOPHYSICAL MODELS FOR PREDICTION AND DESIGN OF EUKARYOTIC CHROMATIN STRUCTURE AN

Fiscal Year: 2009

Project Number: 1R01HG004708-01A1

Serial Number: 4708

Funding: NHGRI

Principal Investigator: MOROZOV, ALEXANDRE V

Abstract: DESCRIPTION (provided by applicant): Our long-term objective is to predict how molecular interactions are translated into gene expression in eukaryotic cells. In eukaryotic genomes, transcriptional regulation is strongly affected by nucleosomes which function to compact DNA and to regulate access to it by simple physical occlusion and by providing the substrate for numerous covalent epigenetic tags. We have recently developed a DNA mechanics-based nucleosome model capable of reproducing in vitro free energies of nucleosome formation with high accuracy. We propose to apply this model to predicting nucleosome positions genome-wide, in S.cerevisiae and other model organisms. We will develop descriptions of chromatin structure that incorporate histone octamers competing with other DNA-binding factors for regulatory sequence. Our preliminary results indicate that this competition may be as important for shaping in vivo chromatin structure as intrinsic nucleosome sequence preferences. We will investigate the accuracy of our predictions by exploring the link between nucleosome positions and gene expression in model systems. We propose to construct promoter sequences that incorporate transcription factor binding sites into a computationally designed nucleosome occupancy profile. These constructs will be assayed for levels of gene expression, providing direct insight into the regulatory role played by nucleosomes. In addition, we propose to carry out high-throughput sequencing of nucleosomes reconstituted in vitro on both genomic and chemically synthesized sequences. This data set will allow us to disentangle intrinsic sequence preferences from in vivo effects, and will enable us to improve the purely structure-based DNA mechanics model in a systematic way. Finally, we propose to carry out microarray and large-scale sequencing studies of the dynamic response of chromatin structure to environmental and genetic perturbations. The proposed studies will significantly enhance our understanding of the connection between regulatory DNA sequence, chromatin, and gene expression. PUBLIC HEALTH RELEVANCE: The ability to predict how chromatin structure affects gene expression will open a novel pathway towards numerous applications in biology and medicine, including rational drug design and new, chromatin-based approaches to rewiring cellular networks. The ability to exercise precise transcriptional control over the amount and the type of proteins produced by the cell through making directed changes in chromatin structure will find many uses in bioengineering and synthetic biology, including production of synthetic hormones, enzymes, and therapeutic agents.

COCAINE DISRUPTION OF MATERNAL MOTIVATION: PREFERENCE FOR PUPS VS. COCAINE

Fiscal Year: 2009

Project Number: 1R03DA027945-01

Serial Number: 27945

Funding: NIDA

Principal Investigator: PEREIRA ARBOLEYA, MARIANA

Abstract: DESCRIPTION (provided by applicant): Cocaine abuse in women during the postpartum period is associated with dysfunctional maternal care, which can have severe lifelong consequences for the child. The preclinical model used in the following experiments employs a dual-choice paradigm based on the conditioned place preference procedure to offer postpartum maternal rats a choice between environments associated with pups and cocaine. In some females, cocaine is reinforcing enough to compete with maternal motivation for pups, whereas in others maternal motivation is the stronger, more resilient force. We identified neurons within discrete brain regions, including the medial prefrontal cortex (mPFC), nucleus accumbens (NA) and medial preoptic area/ventral bed nucleus of the stria terminalis (mPOA/vBST) that are differentially activated when postpartum rats express a motivated preference for cocaine- or pup-associated environments. The proposed research project is part of an ongoing research project funded by a NARSAD Young Investigator Award, designed to test the hypothesis that preference for pups versus cocaine-associated environments is regulated by the concerted activity of regionally distributed networks of neurons in the regions mentioned above. We will determine the functional role of these sites in the expression of motivated choices by transiently inactivating (bupivacaine infusion) each of them and then testing stimulus-associated environment preference (Specific Aim I). Immunocytochemical analysis of the early immediate gene product cFos will reveal regional specificity of neuronal inactivation, and consequent changes in neuronal activity within the remaining key components of the motivational circuitry (Specific Aim II). Further, the causal contribution of dopamine (DA) and glutamate (GLU) neurotransmission within key regions to the choice behavior will be determined with site- specific infusion of selective DA and GLU agonists and/or antagonists (Specific Aim III). These experiments will uncover which regions are causally involved in the motivated choice responses of pup- and cocaine-seeking behaviors, and further reveal how these CNS sites interact to promote the choice for one stimulus over the other. PUBLIC HEALTH RELEVANCE: The 2007 National Survey on Drug Use and Health (NSDUH) estimated the rate of cocaine use among pregnant women at 5.2%. Cocaine abuse in women during pregnancy and the postpartum period severely disrupts mother-infant interactions, which can have negative lifelong consequences for both members of the dyad, often including severe mental health consequences. Substance abuse remains a pressing public health problem that affects millions of women and their babies, and imposes enormous financial and social burdens on society. This application is focused in the understanding of the neural substrate that mediates the motivational aspects of maternal behavior during the postpartum period, particularly within the context of the disruptive effects of drugs with high abuse potential such as cocaine. Through a combination of neurobiological and behavioral techniques, this project will uncover which brain regions are causally involved in the motivated choice responses of pup- and cocaine-seeking behaviors, and further reveal how they interact to promote the preference for one stimulus over the other. Stimulant abuse is an insidious and dangerous chronic recurring and relapsing disease. While the human condition is vastly more complex than our preclinical model, our data offer critical information of the fundamental neurobiological underpinning of the disruptive effect of cocaine on maternal motivation. This preclinical information might contribute to the development of strategies for treatments or methods of prevention of this tragic human condition, with particular emphasis in maintaining or restoring mother- infant bonding.

ROLE OF TUMOR AND STROMAL CELL METABOLISM IN STRESS ADAPTATION AND PROGRESSION

Fiscal Year: 2009

Project Number: 1RC1CA147961-01

Serial Number: 147961

Funding: NCI

Principal Investigator: COLLER, HILARY A;RABINOWITZ, JOSHUA D;WHITE, EILEEN ;

Abstract: DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (15) Translational Science and specific Challenge Topic, 15-CA-112: Cancer Cell Energy Metabolism and Cancer Causation The molecular events that covert normal cells into tumor cells alter cellular metabolism to aerobic glycolysis, which favors macromolecular synthesis at the expense of efficient ATP production. This metabolic switch that facilitates tumor growth remains a fundamental distinction between normal and tumor cells that has yet to be effectively exploited of cancer therapy. Deregulated tumor cell growth and altered metabolism are also a source of metabolic stress. Oxygen, nutrient and factor deprivation caused by growth to high density and insufficient angiogenesis, and lactate production from glycolysis, are common features and sources of stress in the tumor microenvironment. How tumor and associated stromal cells respond to this stress effects tumor progression and treatment response. Possible responses include cell death (apoptosis or necrosis), cell cycle exit (quiescence or senescence) or adaptation and survival (autophagy). The catabolic process of autophagy is a lysosomal degradation pathway induced by metabolic stress that confers stress tolerance by maintaining energy homeostasis through cellular self-consumption and recycling, and by mitigating oxidative damage through the degradation of malfunctioning organelles and proteins. Autophagy may be particularly important to cells in a glycolytic state and under conditions of metabolic stress. Thus, the adaptation of tumor and associated stromal cells to metabolic stress is intimately linked to their inherent metabolic activity, but the role of metabolism in the context of tumor-stromal interaction is not known. Moreover, exactly how autophagy contributes to cellular metabolism and survival in the tumor microenvironment is not clear. We hypothesize that metabolic adaptation in tumor tissue allows prolonged survival to stress permitting tumor relapse and that discerning the underlying mechanisms will provide new approaches to cancer therapy. To test this hypothesis, we plan to define tumor and stromal cell metabolism, the role of catabolism through autophagy, and the mechanism of metabolic stress adaptation. Growth to high density induces fibroblasts to alter metabolism, activate autophagy, exit the cell cycle, and enter quiescence. We discovered that induction of quiescence of stromal fibroblasts dramatically alters gene expression, including activation of Notch signaling that is required for cell cycle reentry. Remarkably, glucose consumption and utilization is high in quiescent fibroblasts, which causes lactate secretion which can alter the microenvironment; but how does it influence tumorigenesis is not known. Analogous to quiescence in fibroblasts, apoptosis-defective tumor cells subjected to metabolic stress activate autophagy, exit the cell cycle and enter a state of prolonged dormancy from which they can reenter the cell cycle when growth conditions are favorable. We discovered that tumor cell dormancy dramatically alters the cellular proteome and gene expression with evidence of compensatory nutrient uptake, Foxo and Notch pathway activation, and induction of uncharacterized mammalian homologues of yeast genes required for quiescence induced by carbon source limitation. These findings suggest that major metabolic reprogramming accompanies the transition from proliferation to dormancy to cell cycle reentry. These striking parallels between quiescence and dormancy suggest that they are governed by common metabolic reprogramming events that may be a fundamental aspect of tumor biology that has yet to be explored. We propose to define the metabolic networks and role of metabolism as cells transition from proliferation to dormancy or quiescence to cell cycle reentry and how this alters tumor-stromal interaction. Preventing tumor cells from successful metabolic adaptation to stress enabling sustained dormancy and recovery may provide a novel approach to cancer therapy. Cancer cells acquire mutations that deregulate cell growth that also alter cellular metabolism. It has recently become apparent that the metabolic reprogramming of tumor cells is necessary to provide the building blocks for the macromolecular synthesis of proteins, lipids and nucleic acids necessary to support tumor cell growth. Importantly, these metabolic alterations distinguish tumor cells from normal cells, providing a potential therapeutic window and novel targets for anti-cancer drug discovery. PUBLIC HEALTH RELEVANCE: Cancer cells acquire mutations that deregulate cell growth that also alter cellular metabolism. It has recently become apparent that the metabolic reprogramming of tumor cells is necessary to provide the building blocks for the macromolecular synthesis of proteins, lipids and nucleic acids necessary to support tumor cell growth. Importantly, these metabolic alterations distinguish tumor cells from normal cells, providing a potential therapeutic window and novel targets for anti-cancer drug discovery.

BEHAVIORAL AND GENETIC BIOMARKER DEVELOPMENT FOR AUTISM AND RELATED DISORDERS

Fiscal Year: 2009

Project Number: 1RC1MH088288-01

Serial Number: 88288

Funding: NIMH

Principal Investigator: BRZUSTOWICZ, LINDA M

Abstract: DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (03) Biomarker Discovery and Validation and specific Challenge Topic 03-MH-101*: Biomarkers in Mental Disorders. The overall goal of this proposal is to advance the development of behavioral and genetic biomarkers for autism and related disorders. While it is clear that autism has a strong inherited genetic component, very large scale genetic studies that have relied only on a general diagnosis of autism (spectrum) disorders (or other information only on affected individuals) have had limited success in identifying risk alleles, leaving a critical issue for the field. Clearly, alternative genetic study designs are needed to complement existing studies. Behavioral biomarkers, especially language ability, have been used with some success to increase power in gene mapping, but to date studies have focused on detailed behavioral assessments only of subjects with autism and not their family members, despite an extensive literature defining increased rates of related phenotypes in family members. This critical gap will be filled by our project. We will use our existing family set with an extensive existing database of clinical and genetic data from all family members, where each family contains at least one proband with autism and at least one proband with a language deficit, to define biomarkers for risk. For Aim 1, Behavioral Biomarker Development, we will develop a set of behavioral biomarkers of genetic risk for autism and related disorders. We will analyze our extensive behavioral testing database to determine which measures have the strongest genetic effects and further examine latent class structures for both data reduction and to reduce measurement error. We will conduct follow-up assessments on a subset of study participants to determine longitudinal stability of selected biomarkers. For Aim 2, Behavioral Biomarker Validation, we will validate inherited components of the behavioral biomarkers through the use of genome-wide analysis. We will use analysis of quantitative and dichotomous behavioral biomarker data using a quasi-Bayesian posterior probability method to elucidate the genetic architecture of risk, providing evidence of the nature of the inherited genetic component. For Aim 3, Genetic Biomarker Identification, we will identify specific DNA variations associated with genetic risk for autism and related disorders. We will test both common and rare variants, SNPs and CNVs, from candidate genes within the regions identified in Aim 2 and evaluate additional variants from the literature as potential factors modulating a network of risk-determining genes. Overall, we plan to combine analysis of behavioral and genetic biomarkers to develop more accurate models for the prediction of risk for autism spectrum disorders. Our existing detailed clinical and behavioral information, as well as plans for follow-up assessments, will also provide important preliminary data for future comparative effectiveness studies on elements of clinical course and treatment response related to specific biomarkers. It is hoped that these studies will provide substantive insights into the causes of, and effective treatments for, autism. Autism is a serious and debilitating disorder. While there is strong evidence supporting a significant genetic component to the disorder, the identification of specific susceptibility genes has been difficult. Identification of susceptibility genes through the approaches proposed could provide important insights into biological basis of this illness, which could result in the development of novel treatments.

OPTIMIZING BIOACTIVE HYDROGELS TO CONTROL ANGIOGENESIS AND INFLAMMATION IN FUNCTION

Fiscal Year: 2009

Project Number: 1R21HL091465-01A2

Serial Number: 91465

Funding: NHLBI

Principal Investigator: SUNG, HAK-JOON

Abstract: DESCRIPTION (provided by applicant): Biomedical implants that facilitate communication/interaction with the surrounding tissue and/or circulatory system are rendered ineffective by the huge diffusion barrier and increased electrical resistance presented by the fibrous capsule. Examples of these devices include biomaterial implants, biosensors, implantable drug-delivery devices and tissue-engineering scaffolds. The foreign body response is characterized by enhanced recruitment of inflammatory cells. For successful implants, it is ideal to have the device surrounded and penetrated by highly vascularized tissue. Angiogenesis is the formation of new blood vessels from the existing vascular system. Both angiogenesis and inflammation are inescapable in vivo responses to all biomaterial implants. Most biomaterial implants are designed either to reduce inflammation or to improve vascularization. Although progress has been made, many studies overlook the important interconnectivity of inflammation with angiogenesis and focus on only simple in vitro outcomes. Indeed, eventual clinical success of biomaterials will require coping with the interconnectivity of the complex in vivo environment. There is emerging evidence that inflammatory cells regulate the functions of endothelial cells related to angiogenesis. However, the signals initiating angiogenesis in inflammation are complex and difficult to define. The proposed research addresses the hypothesis: the biomaterial-induced inflammatory response may be critical to control angiogenesis. Elucidating a clear physiological mechanism for angiogenesis in biomaterial-induced inflammation will provide new paradigms of biomaterial design and fabrication for the next generation of biomaterials. In order to test this hypothesis, a new class of biomaterials, hydrogels-fabricated from Polyethylene glycol (PEG)-cross-linked tyrosine-derived polycarbonate, has been synthesized and characterized. The hydrogel scaffolds will be made more bioactive to control inflammation and angiogenesis by introducing functional peptides on the polymers. A series of studies will be performed to investigate the role(s) of inflammation in angiogenesis on the hydrogel scaffolds. This study will have high impact on implantation-targeted biomaterial research, because elucidating a mechanism that initiates host inflammatory responses and the subsequent vascularization of biomaterial implants is high risk but very important. The identification of a clear mechanism will provide an efficient and realistic paradigm of the interconnectivity of inflammation with angiogenesis for the functional survival of biomaterial implants. This study will involve sophisticated bioengineering-based technical challenges, such as development of a new class of biomaterial scaffolds, fabrication of scaffold materials to be bioactive, and a quantitative imaging approach using multiphoton microscopy to measure multiple cell functions. The high risk nature of this work has necessitated several different approaches to generate the scaffolds and modify them for testing of the role of inflammation in angiogenesis and of angiogenesis in implant function and survival. PUBLIC HEALTH RELEVANCE: This project will optimize the inherent ability of the inflammatory process present in all biomaterial implant applications to 1) promote angiogenesis by utilizing bioactive molecules in the implant scaffold and 2) enhance design of the implant to release degradation products that direct angiogenesis. To overcome the difficulties in studying inflammatory and angiogenic responses to biomaterials, several high risk in vitro and in vivo methods will be utilized.

MODULATION OF HUMAN REWARD CIRCUITRY BY SOCIAL FACTORS

Fiscal Year: 2009

Project Number: 1R01MH084081-01A1

Serial Number: 84081

Funding: NIMH

Principal Investigator: DELGADO, MAURICIO R.

Abstract: DESCRIPTION (provided by applicant): Motivated or goal-directed behaviors often abide by simple rules of reinforcement. That is, actions that lead to positive consequences tend to be repeated at a greater frequency, while actions that lead to negative consequences tend to be avoided. Fundamental research in non-human animals, complemented by an array of human neuroimaging studies, has delineated a basic neurocircuitry underlying reward-related learning and motivated behaviors. Such research has specifically identified circuits linking cortical structures and the basal ganglia, particularly its input unit - the striatum, a heterogeneous structure in terms of connectivity and functionality - as an interface for the processing of motor and motivational information. However, it is unclear how these basic mechanisms characterized by simple behaviors (e.g., learning that a button press leads to a reward) extend to more complex motivated behaviors typically displayed in society (e.g., learning that an individual is trustworthy and interactions will lead to rewards). The availability of social information and the prospects of social interactions are factors that challenge current thinking of how the brain processes reward and punishment feedback and how it uses such information to make decisions. The goal of this proposal is to use fMRI to investigate how social factors modulate the basic behavioral and neural mechanisms underlying reward-related processing as a precursor to understanding how social influences on motivated behavior impact well-being and mental health. Specifically, the proposed studies attempt to build on a solid and existing research foundation on the neural circuitry of reward-related processing by first: establishing the behavior and neural correlates of a simple task modeled after animal research and second: adding a social component to a similar task to investigate overlap in mechanisms. This approach allows for a translational method that, in future research, can also be further extended to developmental (e.g., the role of social feedback from a social network during adolescence) and clinical settings (e.g., social feedback during observational learning in autism). The proposed studies will investigate two aims. Specific Aim 1 - We will investigate how socially relevant feedback, such as reward feedback from a social network of peers, is processed in the brain and how it compares to non-social feedback. Specific Aim 2 - We will investigate how socially relevant feedback creates social expectations that influence mechanisms of reward-related learning and if such mechanisms are unaffected by non-social feedback. The studies outlined in the proposal will build on our knowledge of the role of corticostriatal systems during simple motivated behavior to probe the antecedents and consequences of social motivation, as a platform to understand how social behavior breaks down in clinical disorders in the future.

THE ROLE OF GRP AND GRPERGIC CIRCUITRY IN FEAR MEMORY

Fiscal Year: 2009

Project Number: 1R01MH080328-01A2

Serial Number: 80328

Funding: NIMH

Principal Investigator: SHUMYATSKY, GLEB P

Abstract: DESCRIPTION (provided by applicant): Sensory information reaching the brain is processed by region-restricted neural networks leading to learning and adaptation to a constantly changing environment. Recent studies have elucidated some of the general molecular mechanisms underlying learning and memory; however, it becomes increasingly clear that there are gene networks that uniquely position key neural circuits to control specialized behaviors. The long-term goal of this work is to systematically characterize how gene regulatory networks control neural circuits dedicated to learned fear. More specifically, this proposal is focused on a gene, gastrin-releasing peptide (GRP), that this laboratory has previously identified as expressed in the neural circuits specifically involved in processing fear- related conditioned stimulus (CS) information. Recently published work from this laboratory suggests that GRP and neuronal circuits that express GRP have an important role in fear memory. The hypothesis will be tested that GRPergic neural circuitry is involved in unimodal and multimodal types of fear memory. A multidisciplinary approach that combines mammalian genetics, behavior and electrophysiology will be used to address the following Specific Aims. Aim 1 will ask whether GRP in the amygdala regulates fear memory. GRP knockout mice will be analyzed in amygdala synaptic plasticity and memory in fear conditioning. The anatomic site of GRP function will be examined by GRP administration in the amygdala of the GRP knockout mice. Aim 2 will ask whether selective elimination of the GRPergic neurons in two different areas of the amygdala differentially affects unimodal and multimodal memories of fear. Aim 3 will ask whether there are two GRP expressing neural microcircuits in the amygdala processing differentially unimodal and multimodal fear memories. GRP and GRPR can be used to develop drugs, targeting neural circuits dedicated to fear and anxiety, and thus more effective interventions can be envisioned that lack side effects. PUBLIC HEALTH RELEVANCE Gastrin-releasing peptide (GRP) may be used to design drugs for the selective regulation of learned fear- related behaviors. Moreover, GRP knockout mice can be used to study how memory of fear is involved in borderline personality disorder, PTSD and generalized anxiety disorder.

BIOLOGY OF THE NK CELL CYTOLYTIC ACTIVITY RHYTHM

Fiscal Year: 2009

Project Number: 1R01HL088041-01A1

Serial Number: 88041

Funding: NHLBI

Principal Investigator: SARKAR, DIPAK KUMAR

Abstract: Circadian and daily rhythms regulate many aspects of physiology and behavior. Although a growing number of studies suggest that circadian disruptions may render organisms more susceptible to infection and cancer, the molecular links between the circadian system and the immune system are largely unknown. Natural killer (NK) cell cytolytic activity has been shown to follow a daily rhythm, which is under a circadian control. NK cells participate in the immune response against viral infections and cancer. Employing the rat animal model, this proposal determines the mechanisms by which the circadian rhythm of splenic NK cell cytolytic function is regulated. It tests the hypothesis that splenic NK cells are provided with molecular clocks that orchestrate the rhythmic expression of cytolytic factors that drive the circadian rhythm of NK cell cytolytic function. The hypothalamic suprachaismatic nucleus, which is itsetf, entrained by photic input, coordinates the expression of clock regulatory genes by altering the sympathetic outflow to the splenic NK cells. The proposed research uses magnetic separation techniques to isolate the NK cell population from the spleen to determine the changes in cytotoxic factors, cytokines and clock genes in these cells; employs gene knockdown techniques in NK cells to study the influence Of clock genes on cytolytic function; analyzes the circadian variations in the clearance of NK-sensitive tumor cells; determines the susceptibility of NK cell function to jet lag, a common circadian disruption, and uses pharmacological tools to investigate the signaling mechanism critically involved in resetting the circadian clock in splenic and clonal NK cells. The proposed series of studies should enhance our knowledge of the physiological mechanisms orchestrating NK cell circadian function.

EXPERIMENTS & COMPUTATIONS TO FIND AGGREGATION-PRONE ENSEMBLES OF ALPHA-SYNUCLEIN

Fiscal Year: 2009

Project Number: 1R01GM087012-01A1

Serial Number: 87012

Funding: NIGMS

Principal Investigator: BAUM, JEAN S ;LEVY, RONALD ;

Abstract: DESCRIPTION (provided by applicant): a-synuclein (aSyn) is an intrinsically disordered protein that appears in aggregated form in the brains of patients with Parkinson's disease. The conversion of monomer to aggregate is complex. Aggregation rates of aSyn are very sensitive to changes in amino acid sequence and environmental conditions. Understanding aSyn aggregation requires characterizing the ensemble of conformations adopted by the monomer and correlating them to aggregation behavior. Though many hypotheses have been proposed to relate aSyn's aggregation behavior to its interconverting conformational ensembles, a consistent molecular description of the aSyn conformational ensembles and their relationship to aggregation remains elusive. This proposal integrates NMR and computational approaches to characterize and explicitly visualize the intrinsically disordered conformational ensembles of aSyn and the early stages of aggregation under different sequence and environmental conditions. The goal is to identify the elements of transient 2o and/or 3o structure that are key for initiation of aggregation and determine their stabilizing driving forces. Identifying the structural basis of aSyn monomer aggregation propensity may be critical for developing inhibitors for the aggregation steps that precede the toxic aggregation cascade. Once developed, this integrated approach can be applied to other important biological functions or diseases involving intrinsically disordered proteins. PUBLIC HEALTH RELEVANCE: Parkinson's disease is the second most prevalent of the late onset neurodegenerative diseases. a-synuclein, an extremely important protein involved in the etiology of Parkinson's disease will be modeled at the molecular level by integrating NMR and computational approaches. Understanding the role of the monomeric conformational ensembles of a-synuclein may be critical for developing inhibition strategies against amyloid formation.

NEURAL MECHANISMS OF AVOIDANCE LEARNING AND ACTIVE COPING VIA EMOTION REGULATION

Fiscal Year: 2009

Project Number: 1R01DA027764-01

Serial Number: 27764

Funding: NIDA

Principal Investigator: DELGADO, MAURICIO R.

Abstract: DESCRIPTION (provided by applicant): Avoiding potentially negative outcomes is a common practice in human society. For instance, a person may drive to work earlier to avoid the negative consequences and stress associated with rush-hour traffic. In such circumstances, the traffic acts as a negative reinforcer, eliciting negative affect and strengthening the action of driving earlier. Negative reinforcers can have beneficial short-term influences on behavior by prompting one to actively cope with negative affect (e.g. avoiding traffic stress). However, an avoidance response can also become maladaptive if it prevents an individual from directly addressing the stressful event, causing long-term consequences such as fostering an addictive behavior (e.g., avoiding work deadlines and stress with drug use). These responses are difficult to extinguish and can become habitual, creating a burden for the individual and society. Thus, it is imperative to understand how aversive outcomes influence human brain and behavior. Non-human animal models of aversive learning highlight the role of striatal circuits, commonly implicated in reinforcement learning, in the acquisition of avoidance or coping responses (e.g., Salamone, 1994; LeDoux & Gorman, 2001). Yet, less is known about 1) human mechanisms of avoidance learning; 2) alternative means of coping with negative affect that are dependent on higher-order cognition and control and; 3) the influence of a pre-existing negative emotional state or stress on active coping strategies. The goal of this application is to investigate how negative reinforcement influences human brain and behavior as a precursor to understanding how humans learn to cope with potential negative outcomes that can influence decision-making in maladaptive ways (e.g., drug abuse to alleviate negative affect). The proposed studies will build on a solid and existing research foundation on affective learning in animals, which links the striatum and avoidance behavior, by first: establishing in humans the behavior and neural correlates of simple negative reinforcement tasks modeled after animal research; and second: examining alternative means of active coping with negative reinforcement that are more common to humans, such as higher-order cognitive strategies. Finally, proposed studies will examine the influence of a pre-existing acutely stressful state on active coping through avoidance or emotion regulation strategies. This approach allows for a translational method that can also be further extended to developmental (e.g., avoidance learning during adolescence) and clinical settings (e.g., mechanisms of active coping via emotion regulation during drug craving) in future research. PUBLIC HEALTH RELEVANCE: The proposed studies will build on a solid and existing research foundation on affective learning in animals by first: establishing in healthy human brains the behavior and neural correlates of simple negative reinforcement tasks modeled after animal research; and second: examining alternative means of active coping with negative reinforcement that are more common to humans, such as higher-order cognitive strategies. This approach allows for a translational method that can be applied to understanding the relationship between negative reinforcement and substance abuse, setting up future clinical investigations in substance-abusing populations (e.g., mechanisms of active coping via emotion regulation during drug craving).

CRNS: MODEL OF HIPPOCAMPAL-AMYGDALA INTERACTION: IMPLICATIONS FOR PTSD

Fiscal Year: 2009

Project Number: 1R01AA018737-01

Serial Number: 18737

Funding: NIAAA

Principal Investigator: MYERS, CATHERINE E

Abstract: DESCRIPTION (provided by applicant): In response to NSF 08-514 (CRCNS), we propose a new collaborative project to develop a computational model of the interaction of hippocampus, amygdala, and ventromedial prefrontal cortex in conditioning, extinction, and contextual processing. The model will be applied to data collected from patients with post-traumatic stress disorder (PTSD), in the hopes of elucidating the brain substrates of this disorder. Parallel empirical studies, in healthy adults and in patients with PTSD, will be conducted to generate further data to constrain the model, while the model itself will generate new predictions that may drive further empirical studies. We thus anticipate an ongoing process in which the model is revised to account for new data while producing new predictions to guide further empirical studies, which in turn may lead to new diagnostic tools. In addition, the computational model will allow us to investigate the possibility that there may be different subtypes of PTSD that involve different nodes of brain dysfunction contributing to a common symptomatology. The inability to conduct controlled studies of the disorder in humans (e.g. via experimentally-induced trauma), and the resulting focus on individuals who have already developed the disorder, have to some extent hindered research into causes vs. effects of the disorder. Although animal models have had some success, there is also a potential here for computational modeling to examine how damage to or dysfunction of various brain systems (alone or in combination) might contribute to PTSD symptomatology. The project represents a new collaboration among experts on computational neuroscience of the hippocampus in conditioning and contextual processing (Myers), on the structural, functional, and behavioral abnormalities in PTSD (Gilbertson, Orr), and on classical fear conditioning in humans and animal models of anxiety (Servatius). Intellectual Merit The outcome of this research will provide the field with a deeper understanding of the role of several critical brain structures in normal learning and memory, as well as in PTSD. The empirical work will expand our understanding of generalized learning deficits in PTSD, and allow us to test model predictions, as well as producing new data to constrain the model. The project brings together researchers from several disciplines - computational neuroscience, experimental neuropsychology, clinical psychology, psychophysiology, and animal models of human anxiety disorders -- and builds on the expertise of each. The senior personnel have significant prior experience in the research methodologies involved and have strong publication records as well as commitments to teaching and mentoring. Broader Impact PTSD may affect over 6% of the US population at some point during their lifetimes, with accompanying health care burdens as well as societal costs due to lost participation in professional and personal activities. The proposed work will increase our understanding of PTSD and examine the idea that it may not be a unified disorder, but a family of pathologies that share clusters of common symptoms with each other and also with the broader spectrum of anxiety disorders. There will be implications for prevention, through better understanding of pre-existing risk factors, and for optimizing treatment that targets possible PTSD subtypes. Computational modeling and pilot empirical work will take place at Rutgers-Newark, which is consistently ranked as the #1 most diverse public university in the nation. Myers trains 4-6 undergraduate interns each semester, many of whom are underrepresented minorities, often the first in their families to go to college. Three of her current undergraduate interns have presented their research at national scientific conferences. Myers herself is one of the few female researchers represented in the field of computational neuroscience. The program will also fund a graduate student and 1-2 undergraduate researchers, who will play key roles in experimental design, empirical testing, data analysis, and report writing.

VIBRATIONAL SPECTROSCOPY: COLLECTIN INTERACTIONS WITH PHYSIOLOGICAL LIGANDS

Fiscal Year: 2009

Project Number: 1P01AI083222-01A2

Serial Number: 83222

Funding: NIAID

Principal Investigator: MENDELSOHN, RICHARD

Abstract: The long term objective of this proposal is to elucidate the molecular basis by which collectins in the lungprovide the first challenge to airborne pathogens in host defense. The surfactant specific collectins SP-Aand SP-D, through their interactions with lipopolysaccharides(LPS), are responsible in part for this activity.With the technological expertise in novel vibrational spectroscopic appraoches developed in this lab, wepropose two specific aims to characterize the interaction of both proteins with physiologocal LPSderivatives. With the expertise of the other investigators in the program project, we will have access to alarge number of genetic variants of these proteins, permitting us to pinpoint the primary interaction sites in avariety of biologically relevant physical preparations.Two specific aims are proposed. First, since both collectins recognize carbohydrate and other polar ligandsites, we will determine those collectin structural factors important for binding to LPS and its variants inLangmuir films (i.e. monolayers) at the air/water interface using a unique vibrational spectrscopy experiment(Infrared Reflection Absorption Spectroscopy- IRRAS) that extracts molecular structural and orientationalinformation from the monolayer constituents. These films mimic polar regions of bacterial Gram negativeouter membrane monolayers. The integrating hypothesis for this Aim is that at the level of molecularstructure, the interaction between collectins and LPS depends on particular elements of protein structure andemphasizes specific regions of the collectin-carbohydrate recognition domain. Raman microcrystallographycomplements IRRAS and provides very specific structural information about protein side chains involved inthe initial recognition event. In the second Aim, we will complement studies of protein structural changeswith IR experiments that track structural changes both in the acyl chains and in the polar regions of the LPSderivatives. We will monitor collectin/LPS interaction under conditions where Langmuir films are not the onlyreasonable experimental paradigm. Physical states of lipid and lipid/protein complexes from which structuralinformation will be acquired include vesicles, monolayers, supported oriented multibilayers, and micelles.RELEVANCE (See instructions):The collectins play a vital role in host defense against microorganisms, although little is known of themolecular basis for microbial recognition. Following the planned novel vibrational spectroscopic studies, amore detailed understanding of this mechanism of host defense will emerge. Since threats to personalsecurity are currently presumed significant, understanding the recognition mechanisms are important foreffective design of agents for therapeutic intervention

COMBINATORIAL APPROACHES FOR STUDYING MULTIPLE CUES REGULATING HUMAN PLURIPOTENT

Fiscal Year: 2009

Project Number: 1DP2OD006462-01

Serial Number: 6462

Funding: OD

Principal Investigator: LEE, KIBUM

Abstract: DESCRIPTION (Provided by the applicant) Abstract: Human pluripotent stem cells (hPSCs) are promising resources as cell-based therapies for the debilitating injuries caused by many neuro-degenerative diseases. However, controlling hPSC differentiation into lineage-specific neural cells is one of the most important problems needed to be addressed before their potential for neuro-regenerative medicine can be fully realized. A detailed insight into the functions of extracellular microenvironments and intrinsic cellular regulators which dynamically regulate the hPSC neurogenesis into neural/neuronal cells is a prerequisite for addressing the aforementioned challenges. However, functions of hPSC microenvironments are much more complicated to investigate because of our lack of knowledge about the multiple signals inducing differentiation and limited methods available for investigation. Therefore, the primary focus of our study is to develop innovative methods to identify optimal cues for hPSC differentiation into subtype specific neurons and genetic manipulation of hPSCs using non-viral siRNA based transfection tools. Our innovative approaches will allow for the establishment of novel cell-based assay tools and siRNA based genetic manipulation tools for selective and efficient neuro-differentiation of hPSCs. Moreover, efforts will be made to integrate these studies into one multianalytic microfluidics platform for synchronized control of microenvironmental cues and intrinsic cellular regulators synergistically. The PI's research experiences in nanoscale biomaterials, functional genomics, and stem cell biology and current interdisciplinary research programs aiming at investigating cellular interactions within microenvironments would be critical to develop the aforementioned/innovative tools. Public Health Relevance: Neuro-degenerative diseases (e.g. Alzheimer's disease and Parkinson's disease) and spinal cord injury, affects about a few million Americans, who experience life-long debilitating paralysis or even death due to the injury. Therefore, there is an urgent need for the development of cell-based therapies for neuro-regenerative medicine, where human pluripotent stem cells (hPSCs) are extremely promising resources for transplantation therapies as they possess the unique ability to self-renew and give rise to all somatic cell lineages. Goals of this proposed study is to develop innovative methods for identifying as well as understanding the temporal/spatial effects from microenvironmental cues and intrinsic cellular programs on growth, differentiation, and molecular specification of human pluripotent stem cell (hPSC) differentiation into sub-type specific neurons.

EXPLORING GENETIC INFLUENCES ON ALCOHOL USE USING NOVEL STATISTICAL METHODS

PI: BUCKMAN, JENNIFER jbuckman@rci.rutgers.edu

7/10/09

Abstract: The training and research plans of this second revised K01 application will directly advance the candidate's long term career goal of developing an independent, transdisciplinary line of research that investigates subtypes of risk for alcohol-related problems and of alcohol use disorders based on phenotypic and genotypic information. Career development is sought in statistical genetics (related to hypothesis generation and testing), molecular genetics, and their integration with alcohol studies. These training objectives will be accomplished through coursework, workshops, seminars, and conferences; through extensive mentoring and consulting with senior investigators whose research is directly relevant to this application; and through development of collaborations. These activities will provide the foundation for the research plan, which combines nontraditional quantitative methods with an intermediate phenotype approach to generate conceptually- and empirically-based hypotheses about neurogenetic influences on physiological processes using highly informative yet small scale.

NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM Grant Number: 1K01AA017473-01A209

CENTER OF ALCOHOL STUDIES

Rutgers, The State University of New Jersey

Regulation of Lens Fiber Cell Organization

PI: ZHOU, RENPING rzhou@rci.rutgers.edu
1-Jul-09


Abstract: Lens transparency is made possible by a combination of the highly ordered organization of the lens fiber cells, their unique refractive index, and the lack of organelles in the fiber cells. The highly ordered arrangement of the lens fiber cells is critical for proper light transmission, and disruption of this structure by alterations of cell-cell interactions is likely to lead to cataracts. However, signals that regulate lens fiber cell interaction remain largely unknown. Our preliminary studies have identified a new class of molecules, the Eph tyrosine kinase receptor family that regulates lens cell organization. Inactivation of ephrin-A5, a ligand of the Eph receptors, leads to the disruption of N-cadherin localization, change in lens fiber cell shape, disorganization of lens cells, and the development of cataracts. We hypothesize that ephrin-A5, interacting with its receptor(s), regulates N-cadherin-mediated fiber cell adhesion to maintain proper lens cell organization. To test this hypothesis, we will: (1) Examine the spatial and temporal characteristics of the ephrin-A5-null lens, determine when and where during development the lens defects first occur, and whether the loss of ephrin-A5 results in disruptions of lens fiber cell differentiation. The morphology of the lens at different developmental stages will be analyzed using both light and electron microscope techniques. Antibodies against markers of lens cell differentiation will be used in immunohistochemical experiments to study the expression of differentiation markers. (2) Elucidate receptor mechanisms of ephrin-A5 in lens development by examining which specific Eph receptors are expressed in the developing lens and where they are expressed, using Real-Time PCR, in situ hybridization, and immunohistochemistry. Since the interaction between Eph receptors and ligands leads to bidirectional signaling, we plan to analyze whether receptor- mediated signaling, the ligand-mediated reverse signaling or both are required for lens development using selective inactivation of different receptor domains. (3) Study the molecular alterations that lead to cataracts in ephrin-A5-null mice. Preliminary studies have revealed a disruption of N-cadherin distribution in the lens fiber cells. We will determine whether ephrin-A5 receptors interact physically with adherens junction molecules, and analyze effects of the ligand on N-cadherin functions. To critically evaluate roles of N-cadherin in mediating ephrin-A5 function and lens cell adhesion, we plan also to examine expression of a N-cadherin-2-catenin fusion protein in a phenotypic rescue experiment. The proposed studies will establish roles of a previously unsuspected family of molecules in lens development and reveal novel regulations of N-cadherin functions. These studies will enhance our understanding of how lens cell interaction is regulated to ensure lens transparency and provide insights into the mechanisms of cataractogenesis. PUBLIC HEALTH RELEVANCE: Cataracts are a leading cause of blindness. The molecular mechanisms underlie cataractogenesis are incompletely understood. The proposed studies will elucidate molecular mechanisms by which defects in ephrin-A5 signaling lead to cataracts and provide insights into future prevention and treatment of human cataracts.



NATIONAL EYE INSTITUTE Grant Number: 1R03CA141756-0109


CHEMICAL BIOLOGY


Rutgers, The State University of New Jersey

Tocopherol-mediated inhibition of breast carcinogenesis

PI: SUH, NANJOO nsuh@rci.rutgers.edu
1-Jul-09

Abstract: Recent biomedical technology advances have made it possible to detect breast cancer earlier, but options currently available to help patients with primary breast cancer or women at high risk of breast cancer are very limited. The long-term objective of our project is to identify and develop safe and efficacious agents, such as tocopherols, for prevention of breast cancer. Tocopherols are lipophilic phenolic antioxidants present in significant amounts in vegetable oils, such as soybean, corn, canola and cottonseed. Our laboratory has found that mixed tocopherols containing mainly gamma- and delta-tocopherols prevent an estrogen receptor (ER)- positive breast cancer. Based on our preliminary studies, we hypothesize that gamma- and delta-tocopherols, not alpha-tocopherol (known as a classic vitamin E), prevent breast cancer by inhibiting cell proliferation, inflammation, and oxidative stress, and by regulating receptor signaling pathways including peroxisome proliferator activated receptor (PPAR), ER and Her-2. The major specific aims of this project are to characterize the cancer preventive activities of mixed tocopherols as well as pure isoforms of tocopherols since it is not clear which tocopherol isomers are responsible for chemopreventive effects against breast cancer. Using two distinct preclinical models of breast cancer, we will determine the preventive efficacy of mixed tocopherols as well as the pure alpha-, gamma- and delta-tocopherols in an estrogen receptor (ER)-positive breast cancer and in an ER-negative/Her-2 positive breast cancer. Using immunohistochemistry, ELISA, Western Blot analysis and real time RT-PCR, we will investigate the efficacy and safety of tocopherols in animal models of breast cancer, analyze molecular markers and identify the mechanism of action of tocopherols in vivo. The proposed study will determine (a) the preventive efficacy of mixed tocopherols and the pure alpha-, gamma- and delta tocopherols individually in an ER-positive breast cancer model, (b) the preventive efficacy of mixed tocopherols and the pure alpha-, gamma- and delta tocopherols individually in a Her-2 positive breast cancer model using MMTV-Her-2/neu transgenic mice, and (c) the molecular mechanisms of the tocopherols in receptor signaling pathways such as PPAR, ER and Her-2. This project will provide a thorough understanding of cancer preventive activities of different tocopherols and their combinations, elucidate the detailed molecular mechanisms of cancer preventive actions on tocopherols, and provide fundamental information of tocopherols for future human breast cancer prevention studies.

Relevance to public health" In year 2008 alone, approximately 180,000 women are being diagnosed with breast cancer in the United States. Although significant advances in technology have made it possible to detect breast cancer earlier, women at high risk or breast cancer patients after mastectomy or lumpectomy have very few options currently available for the prevention of the disease. Our project is focused on the identification of chemopreventive components available in the diet. The major task of the project is to characterize the cancer preventive activities of tocopherol isomers in an estrogen receptor (ER)-positive breast cancer and an ER-negative/Her-2 positive breast cancer. Our study attempts to understand the mechanism of action of pure tocopherol isomers in the prevention of the disease. The successful achievement of this project may contribute to an understanding of how breast cancer can be prevented by tocopherols before it progresses to its terminal invasive and metastatic stages. Since preclinical data are required for further translation studies to develop new agents for prevention and treatment of breast cancer in humans, our proposed study will provide valuable data to benefit patients with primary breast cancer and women at high risk of developing breast cancer. We believe that the outcome from our research will help reduce breast cancer incidence and mortality in the next decade.


NATIONAL CANCER INSTITUTE Grant Number: 1R03CA141756-0109

CHEMICAL BIOLOGY
Rutgers, The State University of New Jersey

Benzene 2009: Health Effects and Mechanisms of Bone Marrow Toxicity. Implications

PI: SNYDER, ROBERT

7/1/09

Abstract: This application requests partial support from the National Institute for Environmental Health Sciences (NIEHS) for an international symposium entitled BENZENE 2009: Health effects, Mechanisms of Toxicity, Implications for t-AML and Application of the Mode of Action Framework, which will be held at the Technical University of Munich, Germany, September 7-11, 2009. Since its inception the NIEHS has been concerned with benzene because of both its occupational and its environmental toxicity. This symposium will devote its discussion to new findings on the mechanism of benzene-induced bone marrow damage. Benzene is a bone marrow toxin and carcinogen (leukemia) and despite attempts to control exposure in more highly developed countries in Europe and in the US, excessive benzene exposure persists in China and other areas of Asia. Reports have been invited on recent epidemiological studies of benzene-induced bone marrow damage. The investigators will explore advances in benzene-related bone marrow patho-physiology, mechanisms of leukemogenesis; the similarities between benzene-induced leukemia and anticancer treatment related leukemia; recent studies on the effects of benzene on the molecular biology of the bone marrow; benzene exposure; biomarkers such as chromosome damage; and risk assessment. They will make use of the Mode of Action framework when evaluating recent data on benzene and suggest data gaps which when filled will enhance the process of risk assessment. A panel discussion will deal with the issues raised at the meeting and special attention will be paid to the question of the adequacy of current occupational standards to protect workers. Speakers from 13 countries have been invited and special arrangements are being made to bring students from underdeveloped countries to participate. The proceedings will be published as a special issue of Chemico-Biological Interactions. The Symposium will explore recent advances in benzene-related bone marrow patho-physiology, mechanisms of leukemogenesis, epidemiology of benzene-induced bone marrow damage, recent studies on the effects of benzene on the molecular biology of the bone marrow, benzene exposure and biomarkers and risk assessment. The primary interests of NIEHS is in the environmental impacts of exposure to benzene and in fundamental mechanisms of benzene toxicity and how these data can be utilized in benzene risk assessment. Much of the discussion will relate directly to interests of the NIEHS.


NATIONAL INSTITUTE OF ENVIRONMENTAL HEALTH SCIENCES Grant Number: 1R13ES017587-0109


PHARMACOLOGY AND TOXICOLOGY

Rutgers, The State University of New Jersey

Persistence and Desistence in Heavy Drinking and Violence

PI: WHITE, HELENE hewhite@rci.rutgers.edu

6/1/2009

Abstract: The proposed longitudinal study focuses on persistence and desistance in heavy drinking and violence from early adolescence into adulthood among a high-risk group of White and African American men from the Pittsburgh Youth Study (PYS). We plan to collect one additional wave of data in adulthood (approximate age 33-35) from a sample of 476 men who had been interviewed at least annually from approximately age 13 to age 25. The additional assessment will provide needed data to identify which participants desist in violence and heavy drinking and to examine the consequences of early patterns of violence and heavy drinking on adult intimate partner violence and other forms of criminal violence (both under and not under the influence of alcohol) and on positive and negative adult adjustment. Compared to other longitudinal community studies, the PYS has several advantages: sufficient numbers of individuals who have engaged in violence and heavy drinking, official reports of violence and diagnostic measures of alcohol use disorders in addition to self and collateral reports, a large sample of African Americans to allow for racial comparisons, and regular assessments with low attrition. The specific aims are: 1) To delineate the developmental courses of heavy drinking and violence and their co-occurrence from early adolescence to adulthood; 2) To test a model of desistance from heavy drinking and/or violence based on the dynamic interplay among structural and demographic factors, adolescent risk and protective factors, factors in adulthood that hinder or promote desistance, and adult opportunities and life events; and 3) To determine how developmental pathways of heavy drinking and violence from adolescence through young adulthood affect positive and negative outcomes in adulthood. This will be one of the rare studies that will be able to link prospectively assessed developmental patterns of violence and heavy drinking to proximal alcohol-related violence in adulthood. Participants and their intimate partners will be interviewed. We will use growth mixture modeling to examine trajectories of violence and heavy drinking. The groups derived from these analyses, as well as conceptually-developed groups, will be used as outcomes in multinomial logistic regression analyses to examine mediation and moderation models of the desistance process. These groups will also be used as predictors of adult outcomes. Growth mixture modeling will also be used to examine whether there are differences across Whites and African Americans in heavy drinking and violence trajectories. In addition, African Americans and Whites will be tested as a priori groups in multi-group analysis to see if race moderates the proposed causal pathways to desistance. To reduce the costs of alcohol-related violence to society, and to develop interventions that address a broader range of critical factors, it is important that we better understand the processes and mechanisms that lead to desistance from both heavy drinking and violence. PUBLIC HEALTH RELEVANCE: The proposed study has high social relevance in that to reduce the costs of alcohol- related violence to society, it is important that we understand the risk and protective factors that lead to desistance from heavy drinking and violence. If we identify early risk and protective factors that differentiate those who will mature out from those who will not, then we will be better able to target the most high-risk youths for early interventions and to develop more effective and efficient interventions at appropriate developmental stages.

NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM Grant Number: 1R01AA016798-01A1

Thesaurus Terms:

CENTER OF ALCOHOL STUDIES

Rutgers, The State University of New Jersey

Neurophysiology of cocaine-seeking behavior

PI: ROOT, DAVID

5/25/2009

Abstract:

NATIONAL INSTITUTE ON DRUG ABUSE Grant Number: 1F31DA026252-01

Thesaurus Terms:

PSYCHOLOGY

Rutgers, The State University of New Jersey

Pavlovian influences on reward-seeking behavior

PI: SHIFLETT, MICHAEL

5/15/2009

Abstract:

NATIONAL INSTITUTE ON DRUG ABUSE Grant Number: 1R03DA026559-01

Thesaurus Terms:

PSYCHOLOGY

Rutgers, The State University of New Jersey

C. elegans as a model for ricin intoxication

PI: TUMER, NILGUN tumer@aesop.rutgers.edu

5/5/2009

Abstract: Ricin is a plant toxin produced by the castor bean (Ricinus communis) that is extremely toxic to mammalian cells and is classified as a select agent by the US Department of Health and Human Services (HHS) and a category B priority pathogen by the Center for Disease Control (CDC). Although there have been attempts to generate an effective vaccine against ricin, no recombinant vaccine has yet been approved for human use and there are no effective treatment measures against ricin exposure. Therefore there is an urgent need for therapeutics to protect ricin-exposed individuals. To gain understanding into the mechanism of ricin induced cell death, we will take advantage of the unique experimental features of C. elegans that have made it a leading model organism in nearly all areas of biological research. Our primary objective in this proposal is to establish C. elegans as a new whole animal model to study ricin intoxication and to translate our findings to mammals. In preliminary studies, we fed C. elegans with either E. coli expressing ricin A-chain (RTA) or recombinant RTA purified from E. coli and demonstrated that RTA is lethal to C. elegans at extremely low doses. In agreement with mammalian data, we showed that ricin uses a clathrin and a raft independent endocytosis pathway to enter the C. elegans intestinal epithelial cells and entry to the early endosome is critical for cytotoxicity. We identified a novel role for the two major phagocytosis/engulfment pathways in mediating ricin sensitivity in the C. elegans intestine. These results provided support for the development of C. elegans as a new multicellular model for molecular genetic dissection of the mechanisms of ricin transport and cell death. We will examine transport of ricin in the worm intestine, a classic polarized epithelium, in the context of a living animal. C. elegans mutants defective in intracellular trafficking pathways will be used to validate C. elegans as a relevant model for mechanistic analysis of ricin uptake and transport and to identify critical steps in ricin transport that lead to cytotoxicity. We will exploit powerful features of C. elegans genetics to isolate new mutants defective in ricin intoxication. The proposed studies will lead to elucidation of the molecular details of ricin transport and will lay the foundation for identification of inhibitors that can prevent ricin induced cell death. PUBLIC HEALTH RELEVANCE: Our primary objective in this application is to establish C. elegans as a new model to study the mechanism of ricin intoxication and to translate our findings to humans. These studies are significant because currently there are no vaccines, antidotes or any other treatment measures against ricin exposure. These studies will lay the foundation for identification of inhibitors that can protect ricin exposed individuals.

NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 1R21AI077805-01A1

Thesaurus Terms:

BIOTECH CTR/AGRIC & ENVIRON

Rutgers, The State University of New Jersey

Role of Nitric Oxide in Radiation-Induced Lung Injury

PI: MALAVIYA, RAMA rmalaviya@verizon.net

5/1/2009

Abstract: This mentored career development plan will help the P.I. re-enter the field of biomedical research after a 3- year hiatus, and will prepare her to conduct independent research focusing on pulmonary disease pathogenesis and treatment. The plan includes training in pulmonary toxicity, inflammation, and drug delivery through mentored research and educational activities. This experience will enable the P.I. to achieve her overall goal of becoming an independent investigator. The overall goal of the research is to elucidate inflammatory mechanisms mediating lung disease with a particular emphasis on the role of reactive nitrogen species (RONS) in disease pathogenesis. Although RONS have been shown to play a key role in acute lung injury and disease, their role in pneumonitis and consequent fibrosis is unknown and represents the focus of the research. To induce pneumonitis, a mouse model of radiation-induced lung injury will be used. The research plan will test the hypothesis that inhibiting production of RONS immediately following radiation exposure, while not reducing acute injury during the early latent phase of radiation induced injury, will significantly mitigate pneumonitis and consequent fibrosis. For these studies 1400W, a highly specific inhibitor of inducible nitric oxide synthase (iNOS), which has previously been shown to be effective in ameliorating acute lung injury in rodent models will be used. In aim 1, the efficacy of 1400W administered by Alzet micro-osmotic pumps in mitigating radiation induced iNOS activation and RONS generation will be assessed. Measurements will be made of markers of acute lung injury and oxidative/nitrosative stress. Aim 2 will be focused on assessing the effectiveness of 1400W in reducing consequent pneumonitis and fibrosis. In Aim 3, we will optimize a novel nanogel microparticle system to selectively deliver 1400W to the pulmonary vascular compartment of mice after radiation exposure. The results of these studies will provide important mechanistic information on the role of RONS in radiation induced lung injury and may provide clues on a novel efficacious treatment protocol for mitigating lung pneumonitis and fibrosis. RELEVANCE (See instructions): Pneumonitis and fibrosis consequent to acute lung injury are of major health concern. The identification of mechanisms leading to these pathologies and effective treatment protocols is key to minimizing chronic lung injury. (End of Abstract)

NATIONAL HEART, LUNG, AND BLOOD INSTITUTE Grant Number: 1K01HL096426-01

Thesaurus Terms:

PHARMACOLOGY AND TOXICOLOGY

Rutgers, The State University of New Jersey

Creating a Tool to Analyze Protein Sequences Based on Important Amino Acids

PI: PALENCHAR, PETER peterpal@crab.rutgers.edu

4/3/2009

Abstract: Understanding enzyme function is a key component to many public health issues. These include identifying new drug targets for infectious diseases, understanding how mutations in enzyme encoding genes lead to diseases, and identifying enzymes involvedin producing natural products of interest. One limitation to genome-wide analysis (e. g. microarrays) and many biological databases is that they are dependent upon the correct annotation of protein function. In fact, whole systems have been designed to analyze microarray data based mainly on predicted protein function, and while gene annotation has improved, annotation errors still exist. Efforts to improve gene annotation will improve the quality and efficiency of research in the biomedical sciences. This research will generate an ontology to describe how enzymes interact with their substrates and other small molecules (e.g. allosteric regulators). A database will store the functionally important amino acids of specific enzymes and how those amino acids are involved in substrate binding and other types of interactions based on the created ontology. The objective of the database is to create a system that will allow the function of all of the known important amino acids in an enzyme to be described in a detailed but organized manner. Queries of the database then can be performed in a number of methods: 1. If given a protein sequence with a proposed function, it can determine which of the amino acids known to be critical for that function are conserved. 2. Identify all of the enzymes that use a specific amino acid to interact with a specific part of a given molecule (e.g. all of the enzymes that use a lysine to interact with the phosphates of ATP). 3. Using basic statistical approaches, queries such as "Do a significant number of enzymes that bind to ATP use a lysine to interact with the ATP?" can be performed. These types of queries have been employed to mine data from gene ontologies and have helped to revolutionize biology. This proposed research will apply these powerful principles to biochemistry and enzymology for the first time. In addition, this database system will allow the broader biomedical research community to easily identify and understand the importance of critical amino acids in enzymes of interest. PUBLIC HEALTH RELEVANCE: The biochemical understanding of how enzymes function is a key component to public health as mutations in enzyme encoding genes can cause diseases and enzymes are frequently drug targets. Creation of the Functional amino Acid Navigator (FAN) database will allow complex queries to be performed regarding how enzymes function, and in general, will allow more efficient use of time as information from decades of research on enzymes will be collected and stored in an easily queried format.

NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES Grant Number: 1R15GM079792-01A1

Thesaurus Terms:

CHEMISTRY

Rutgers, The State University of New Jersey

2009 Summer Bioengineering Conference

PI: LANGRANA, NOSHIR langrana@rutgers.edu

4/1/2009

Abstract: DESCRIPTION (provided by the applicant): The ASME Summer Bioengineering Conference brings together researchers and educators from a wide variety of biomechanics disciplines to share research progress and stimulate new developments. Topics range from musculoskeletal to cardiovascular, and include studies that go down to the cellular and molecular level. A strong component of this conference is the emphasis on student participation, including specific oral and poster sessions, and encouragement of formal and informal interaction with more senior researchers. The American Society of Mechanical Engineers Bioengineering Division hosts the annual Summer Bioengineering Conference with the general aim of providing a forum for bioengineering researchers and educators to present and discuss current trends in bioengineering over a wide range of subtopics. The uniting theme of the conference is biomechanics, encompassing molecular to tissue and organism-level approaches. The 2009 Summer Bioengineering Conference, to be held June 17-21 in Lake Tahoe, California, will be the 11th installment of this conference, and will feature two invited speakers, numerous workshops, podium sessions, poster sessions, a sponsor/exhibitor area, career fair, and specially highlighted student paper competitions at the BS, MS, and PhD levels. Student participation at this meeting has historically been excellent. Over half of the attendee population at the 2008 conference was students. Our Specific Aim is to maintain or increase student participation through a separate Call for Papers, and by reducing registration fees by $100 for 350 students. A distinguishing feature of this conference is its relaxed atmosphere, promoted through our venue selection, casual dress, and some unscheduled time to permit informal interactions and exchange of ideas among new and experienced participants. By judicious selection of keynote speakers and workshops, we will offer students opportunities to experience emerging biomedical technologies that apply across disciplines. The ASME Summer Bioengineering Conference brings together researchers and educators from a wide variety of biomechanics disciplines to share research progress and stimulate new developments. Topics range from musculoskeletal to cardiovascular, and include studies that go down to the cellular and molecular level. A strong component of this conference is the emphasis on student participation, including specific oral and poster sessions, and encouragement of formal and informal interaction with more senior researchers.

NATIONAL INSTITUTE OF BIOMEDICAL IMAGING AND BIOENGINEERING Grant Number: 1R13EB009564-01

Thesaurus Terms:

BIOMEDICAL ENGINEERING

Rutgers, The State University of New Jersey

Optimizing Bioactive Hydrogels to Control Angiogenesis and Inflammation in Functi

PI: SUNG, HAK-JOON sung@biology.rutgers.edu

4/1/2009

Abstract: Biomedical implants that facilitate communication/interaction with the surrounding tissue and/or circulatory system are rendered ineffective by the huge diffusion barrier and increased electrical resistance presented by the fibrous capsule. Examples of these devices include biomaterial implants, biosensors, implantable drug-delivery devices and tissue-engineering scaffolds. The foreign body response is characterized by enhanced recruitment of inflammatory cells. For successful implants, it is ideal to have the device surrounded and penetrated by highly vascularized tissue. Angiogenesis is the formation of new blood vessels from the existing vascular system. Both angiogenesis and inflammation are inescapable in vivo responses to all biomaterial implants. Most biomaterial implants are designed either to reduce inflammation or to improve vascularization. Although progress has been made, many studies overlook the important interconnectivity of inflammation with angiogenesis and focus on only simple in vitro outcomes. Indeed, eventual clinical success of biomaterials will require coping with the interconnectivity of the complex in vivo environment. There is emerging evidence that inflammatory cells regulate the functions of endothelial cells related to angiogenesis. However, the signals initiating angiogenesis in inflammation are complex and difficult to define. The proposed research addresses the hypothesis: the biomaterial-induced inflammatory response may be critical to control angiogenesis. Elucidating a clear physiological mechanism for angiogenesis in biomaterial-induced inflammation will provide new paradigms of biomaterial design and fabrication for the next generation of biomaterials. In order to test this hypothesis, a new class of biomaterials, hydrogels-fabricated from Polyethylene glycol (PEG)-cross-linked tyrosine-derived polycarbonate, has been synthesized and characterized. The hydrogel scaffolds will be made more bioactive to control inflammation and angiogenesis by introducing functional peptides on the polymers. A series of studies will be performed to investigate the role(s) of inflammation in angiogenesis on the hydrogel scaffolds. This study will have high impact on implantation-targeted biomaterial research, because elucidating a mechanism that initiates host inflammatory responses and the subsequent vascularization of biomaterial implants is high risk but very important. The identification of a clear mechanism will provide an efficient and realistic paradigm of the interconnectivity of inflammation with angiogenesis for the functional survival of biomaterial implants. This study will involve sophisticated bioengineering-based technical challenges, such as development of a new class of biomaterial scaffolds, fabrication of scaffold materials to be bioactive, and a quantitative imaging approach using multiphoton microscopy to measure multiple cell functions. The high risk nature of this work has necessitated several different approaches to generate the scaffolds and modify them for testing of the role of inflammation in angiogenesis and of angiogenesis in implant function and survival. PUBLIC HEALTH RELEVANCE: This project will optimize the inherent ability of the inflammatory process present in all biomaterial implant applications to 1) promote angiogenesis by utilizing bioactive molecules in the implant scaffold and 2) enhance design of the implant to release degradation products that direct angiogenesis. To overcome the difficulties in studying inflammatory and angiogenic responses to biomaterials, several high risk in vitro and in vivo methods will be utilized.

NATIONAL HEART, LUNG, AND BLOOD INSTITUTE Grant Number: 1R21HL091465-01A2

Thesaurus Terms:

CHEMISTRY

Rutgers, The State University of New Jersey

Nanoscale Receptor Blockers for Inhibition of Atherosclerosis

PI: MOGHE, PRABHAS moghe@rci.rutgers.edu

4/1/2009

Abstract: Pathologies in blood vessels arising from the uncontrolled build-up of oxidized lipids contribute to atherosclerosis, a severe cardiovascular disease, which underlies the most common cause of adult death in the U.S. (exceeding one million patients yearly). Few existing therapeutic strategies address the local management of atherogenesis (build-up of oxidized lipids in the blood vessel walls) and related inflammation. The overall goals of this study are to rationally design and characterize nanoscale biomaterials as a novel cell-targeted materials platform for investigating strategies to de-escalate the onset of atherogenesis and reduce accompanying inflammation. The proposed NIH R21 study involves three specific objectives to investigate nanoassembled amphiphilic polymers (NAPs) to maximally inhibit oxidized LDL uptake in human macrophages under physiologic conditions and exhibit potential for specific targeting to inflamed endothelia. Efforts in Aim 1 will involve investigation of innovative designs of nano-assembled amphiphilic polymers (NAP) composition and architecture to promote NAP binding to both SRA-1 and CD36 scavenger receptors on human THP-1 macrophages and thus inhibit uptake of oxidized low-density lipoproteins (oxLDL). New configurations of NAP will tested for improved lipid uptake inhibition in the presence of serum. Studies in Aim 2 will investigate the effect of NAP-scavenger receptor (SR) interactions on the downstream intracellular and cell-secreted intermediates regulating atherogenesis in macrophages, including cytokine secretion; cholesterol ester accumulation; matrix metalloproteinase secretion; and expression analysis of genes involved in pro-atherogenic signaling pathways. Aim 3 is concerned with design and evaluation of the potential of biofunctionalized NAPs to bind to and transport across activated endothelial cell cultures in vitro, thereby creating a simplified in vitro model of the rescue of macrophage cells involved in atherogenesis within the vascular intima. PUBLIC HEALTH RELEVANCE: The excessive uptake of modified forms of LDL in immune blood cells macrophages is one of the hallmarks of fat build-up and vascular disease within blood vessel walls, which can lead to blockage of blood flow, and cause heart disease or stroke. This study will investigate the design of nanoscale assembled polymers with specific architectures, charge displays, and chemistry so as to reduce the uptake of the most damaging forms of lipoproteins within macrophages. The goals of the study are to identify the most effective "nanolipoblocker" configurations that may prevent atherogenesis by targeting activated blood vessel cells and blocking foam cell formation.

NATIONAL HEART, LUNG, AND BLOOD INSTITUTE Grant Number: 1R21HL093753-01A2

Thesaurus Terms:

BIOMEDICAL ENGINEERING

Rutgers, The State University of New Jersey

Multiple Levels of Analysis: Trauma, Violence and PTSD Among Youths'

PI: BOXER, PAUL pboxer@psychology.rutgers.edu

4/1/2009

Abstract: Despite national trends showing declines in violent crime, studies show that violence remains alarmingly high in some communities as well as in homes, schools, and the media. The goal of this proposed R21 project is twofold: To develop a multi-informant, multi-method assessment of youths' exposure to violence in the social environment, anchored to youths' mental health; and to set a foundation for launching new research into the reduction and prevention of the deleterious mental health consequences of youth violence exposure. Our novel approach to assessment is driven by a four-dimensional conception of violence exposure based on the context (setting), content (impact), channel (mode of exposure), and chronicity (frequency of exposure) of specific acts of violence. Our measurement is tied to a comprehensive assessment of mental health (externalizing, internalizing, and traumatic stress disorders) to provide an overall assessment of the burden of violence exposure based on current theory in developmental psychopathology. This investigation will be conducted in socio-economically distressed urban neighborhoods comprising a range of projected risk for exposure to violence in a major metropolitan area of the Northeast. Our study population will be families with children between the ages of 11 and 14 inclusive; we will sample caregiver-youth pairs (dyadic N = 200) for a comprehensive protocol integrating information from the pairs directly with information from data sources across all levels of the social ecology. Our investigation is guided by five aims: 1) Develop and implement a cross-informant, mixed-method protocol for assessing youths' exposure to violence in the social environment; 2) Utilize our novel four-dimensional conception of violence exposure to evaluate the psychometric properties of the new individualized protocol and establish internal validity; 3) Apply a dual pathway model of how exposure to violence affects mental health outcomes to evaluate the criterion validity of the new protocol; 4) Employ multivariate modeling to analyze patterns of exposure and associated outcomes across meaningful subpopulations (e.g., low and high risk for exposure) to assess the generalizability of the new protocol; and 5) Analyze various combinations of reports in order to finalize a protocol for field testing in clinical research settings. Our investigation will provide information critical to understanding how violence exposure leads to negative health outcomes. Ultimately this knowledge will be essential to efforts at reducing the negative impact of exposure to violence. Our approach to measuring youths' exposure to violence is essential for formulating individualized assessment and intervention strategies for ameliorating the negative sequelae of exposure among children and adolescents. Further, the proposed investigation represents a necessary step in expanding our knowledge of how violence in the social ecology impacts individual mental health via biopsychosocial mechanisms. PUBLIC HEALTH RELEVANCE: Although a large body of research confirms detrimental effects of youths' exposure to violence in multiple social contexts on their mental health status, there have been no systematic efforts to develop a theoretically informed measure of exposure to violence across settings. Properly assessing exposure to violence along key dimensions including context as well as severity, frequency, and mode of exposure is an essential step in advancing our understanding of how violence affects youths' mental health outcomes. Better measurement of exposure also will translate into more precise and ultimately more effective intervention strategies. The proposed project aims to develop a new protocol for assessing youths' exposure to violence and exploring how different profiles of exposure account for different patterns of psychopathology.

NATIONAL INSTITUTE OF MENTAL HEALTH Grant Number: 1R21MH085209-01A1

Thesaurus Terms:

PSYCHOLOGY

Rutgers, The State University of New Jersey

Role of intercalated amygdala neurons in the extinction of conditioned fear

PI: PARE, DENIS pare@andromeda.rutgers.edu

3/19/2009

Abstract: Research on the circuits mediating the acquisition of conditioned fear responses constitutes our best hope of understanding human anxiety disorders. The model typically used to study this process is classical fear conditioning where a neutral sensory stimulus (CS) acquires the ability to elicit fear responses after pairing to a noxious stimulus. However, perhaps more important from a clinical perspective is to understand how fear responses subside. Experimentally, this extinction process is modeled with repetitive presentations of the CS alone, resulting in the decline of conditioned fear to control levels. This approach is similar to that used to treat human phobias where subjects are presented with the feared object in the absence of danger. Extinction is known to result from a new learning, which takes place in the amygdala, and competes with the original fear memory to prevent the expression of conditioned fear. However, the mechanisms underlying this new extinction learning remain unclear. This proposal tests the hypothesis that the intercalated (ITC) neurons of the amygdala mediate extinction. The acquisition of conditioned fear is known to involve a potentiation of CS inputs to the basolateral amygdala (BLA). In turn, BLA cells excite more neurons in the central amygdala (CE), which, via their projections to the brainstem and hypothalamus, evoke fear responses. We focus on ITC neurons because they can control the impact of BLA inputs on CE neurons and hence the expression of conditioned fear. Indeed, ITC cells are GABAergic, they receive glutamatergic inputs from BLA, and they generate feed-forward inhibition in CE. Moreover, BLA inputs to ITC neurons can undergo NMDA-dependent LTP. Last, ITC neurons receive a heavy projection from the infralimbic cortex, a cortical area thought to play a critical role in extinction. This leads us to hypothesize that extinction results from an NMDA-dependent potentiation of BLA synapses conveying CS information to ITC neurons, leading to a decreased responsiveness of CE cells to BL inputs about the CS. To test the hypothesis, we will first examine whether extinction is associated with a potentiation of BLA inputs to ITC cells by comparing the amplitude of BLA-evoked responses in ITC neurons recorded with the patch method in slices obtained from rats that underwent fear conditioning only vs. rats that underwent fear conditioning and extinction. Next, we will perform extracellular recordings of ITC cells during fear conditioning, extinction training, and extinction recall, and ask do ITC neurons become more responsive to the CS as a result of extinction training, as predicted by our model. Finally, to test whether ITC cells mediate the influence of the infralimbic cortex on extinction, we will study the responses of extracellularly recorded ITC neurons to infralimbic stimuli and test whether the nature, latency, and duration of evoked responses are compatible with the idea that ITC neurons generate the inhibition of CE neurons by IL stimuli. PUBLIC HEALTH RELEVANCE: Although anxiety disorders affect close to 13% of the population, most available pharmacological treatments have a limited efficacy and entail important side effects. It is thus imperative that we improve our understanding of the mechanisms underlying anxiety disorders to design better treatment strategies. If supported, the hypothesis tested here would open new strategies for the treatment of anxiety disorders.

NATIONAL INSTITUTE OF MENTAL HEALTH Grant Number: 1R01MH083710-01A109

Thesaurus Terms:

Rutgers, The State University of New Jersey

Interactive Effects of Environmental Risk and Prenatal Cocaine Exposure in Preado

PI: WANG, YIPING yipwang@rci.rutgers.edu

3/15/2009

Abstract: This resubmission of K01 DA024113-01 outlines a five-year research career development program to enhance Dr. Wang's initial success in making a major career shift from a successful administrator of intervention development to an independent scientist in substance abuse epidemiology research. The candidate has a track record of success in developing innovative programs that cross-fertilize the fields of substance abuse treatment and early intervention. For the past two years, the candidate has gained the basic skills in theory, research and publication necessary to move toward a career goal as a researcher. The proposed K01 Award serves as a training vehicle to enable the candidate to achieve independence in substance abuse epidemiological research, focusing on the understudied relation between environmental risk and prenatal cocaine exposure and its impact on development in preadolescence. The objectives of the development training are to develop: (1) advanced knowledge of developmental epidemiology of "at risk" populations, (2) advanced skills in research and statistical strategies with longitudinal studies, and (3) academic skills in scientific communication. The candidate will be supported by two premier research institutes: The Center of Alcohol Studies at Rutgers University and the Institute for the Study of Child Development (ISCD) at Robert Wood Johnson Medical School. Dr. Michael Lewis (Director of ISCD), a renowned scholar in developmental research of at risk children, will be the primary mentor. The proposed study will use the transactional multi-risk model to address Specific Aims: (i) to characterize the nature of environmental risk; (ii) to examine developmental outcomes at age 10 as a function of environmental risk and prenatal cocaine exposure; and (iii) to delineate mediating and moderating effects of environmental risk on developmental outcomes in preadolescence as a function of prenatal cocaine exposure. It will use NIDA funded 15-year study on prenatal cocaine exposure (Dr. Lewis, PI) to explore this conceptual model. The completion of this K01 Award will lead the candidate to secure NIDA R01 grants as PI to test the relations between the pattern of change in environmental risk, the developmental pathways of behavioral regulation, and risk taking and substance use behaviors in early adolescence. This research has significant implications for improving intervention by identifying at-risk environments and thus to improve pubic health.

NATIONAL INSTITUTE ON DRUG ABUSE Grant Number: 1K01DA024113-01A1

Thesaurus Terms:

CENTER OF ALCOHOL STUDIES

Rutgers, The State University of New Jersey

LOH at BRCA1/BRCA2 Loci in Mutation Carriers as a Precursor to Breast Tumorigenes

PI: BRENNEMAN, MARK brenneman@biology.rutgers.edu

3/1/2009

Abstract: Women who carry mutations in the BRCA1 or BRCA2 tumor suppressor genes are at extreme risk for breast cancer. BRCA1 or BRCA2 mutation carriers are heterozygous, but their breast tumors typically show loss of heterozygosity (LOH), i.e., loss of the wild-type BRCA allele. LOH at the BRCA loci has been thought of as a rare event, occurring in single cells that are the proximate precursors of a tumor. Recent evidence suggests it may occur instead as earlier or more frequent events giving rise to a large population of cells poised for neoplastic transformation - a "field effect". This study will determine whether LOH at the BRCA loci is typically present in histologically normal or pre-neoplastic breast epithelia of mutation carriers. It is a first step in exploring the potential of LOH at the BRCA loci as a marker for diagnosis or prognosis in mutation carriers, and for new approaches to chemotherapy and chemoprevention that target the DNA repair defects of cells lacking BRCA1 or BRCA2 function. Specific aims will be: 1. To determine whether LOH at the BRCA loci is typically present in pre-neoplastic and histologically normal tissue outside the margins of breast tumors in mutation carriers. Samples of breast tissue will be micro-dissected and assayed by Pyrosequencing, a high-throughput, micro-sequencing technology for mutation detection and allele quantification. For each patient, an individualized Pyrosequencing assay at the mutation site will be designed for direct quantification of the mutant and wild-type alleles. 2. To determine whether LOH at the BRCA loci can be similarly detected in breast tissue removed in prophylactic mastectomy from BRCA mutation carriers, in advance of any diagnosis of breast cancer. 3. To test the feasibility of detecting LOH at the BRCA loci in breast biopsies of BRCA mutation carriers. PUBLIC HEALTH RELEVANCE: This project will study genetic (DNA) changes in women from certain families that are prone to breast cancer. Understanding how these changes take place in breast tissue may open new approaches to early diagnosis, treatment and prevention for this special, high-risk group of patients.

NATIONAL CANCER INSTITUTE Grant Number: 1R21CA122795-01A2

Thesaurus Terms:

GENETICS

Rutgers, The State University of New Jersey

Biophysical Models for Prediction and Design of Eukaryotic Chromatin Structure an

PI: MOROZOV, ALEXANDRE morozoa@rockefeller.edu

2/13/2009

Abstract: Our long-term objective is to predict how molecular interactions are translated into gene expression in eukaryotic cells. In eukaryotic genomes, transcriptional regulation is strongly affected by nucleosomes which function to compact DNA and to regulate access to it by simple physical occlusion and by providing the substrate for numerous covalent epigenetic tags. We have recently developed a DNA mechanics-based nucleosome model capable of reproducing in vitro free energies of nucleosome formation with high accuracy. We propose to apply this model to predicting nucleosome positions genome-wide, in S.cerevisiae and other model organisms. We will develop descriptions of chromatin structure that incorporate histone octamers competing with other DNA-binding factors for regulatory sequence. Our preliminary results indicate that this competition may be as important for shaping in vivo chromatin structure as intrinsic nucleosome sequence preferences. We will investigate the accuracy of our predictions by exploring the link between nucleosome positions and gene expression in model systems. We propose to construct promoter sequences that incorporate transcription factor binding sites into a computationally designed nucleosome occupancy profile. These constructs will be assayed for levels of gene expression, providing direct insight into the regulatory role played by nucleosomes. In addition, we propose to carry out high-throughput sequencing of nucleosomes reconstituted in vitro on both genomic and chemically synthesized sequences. This data set will allow us to disentangle intrinsic sequence preferences from in vivo effects, and will enable us to improve the purely structure-based DNA mechanics model in a systematic way. Finally, we propose to carry out microarray and large-scale sequencing studies of the dynamic response of chromatin structure to environmental and genetic perturbations. The proposed studies will significantly enhance our understanding of the connection between regulatory DNA sequence, chromatin, and gene expression. PUBLIC HEALTH RELEVANCE: The ability to predict how chromatin structure affects gene expression will open a novel pathway towards numerous applications in biology and medicine, including rational drug design and new, chromatin-based approaches to rewiring cellular networks. The ability to exercise precise transcriptional control over the amount and the type of proteins produced by the cell through making directed changes in chromatin structure will find many uses in bioengineering and synthetic biology, including production of synthetic hormones, enzymes, and therapeutic agents.

NATIONAL HUMAN GENOME RESEARCH INSTITUTE Grant Number: 1R01HG004708-01A1

Thesaurus Terms:

PHYSICS AND ASTRONOMY

Rutgers, The State University of New Jersey

ROLE OF EPHRIN-A5 IN LENS FIBER CELL ORGANIZATION

PI: SON, ALEXANDER

2/1/2009

Abstract:

NATIONAL INSTITUTE ON AGING Grant Number: 1F31AG032806-01

Thesaurus Terms:

CHEMICAL BIOLOGY

Rutgers, The State University of New Jersey

Reducing Urban Women's HIV Risk: Soap Opera Videos on Video Capable Cell Phones

PI: JONES, RACHEL racjones@rutgers.edu

12/1/2008

Abstract: Reducing Urban Women's HIV Risk: Soap Opera Videos on Video-Capable Cell Phones According to the CDC, the majority (80%) of women with HIV became infected by unprotected sex with an infected male partner. Black and Latina women accounted for 82% of AIDS diagnoses in women in 2005. The purpose of this proposal is to evaluate whether a series of 12-weekly data and theory-based urban soap operas will promote a reduction in HIV sex risk behavior in young urban African American and Latina women. Toni's Story is based on content analysis of focus group discussions with the target population. By grounding stories in urban women's own experiences, via a popular medium of the soap opera, women can identify with the heroines' emotionally charged process of change, transforming their behavior through a new awareness of their value as women, of their choices, and their potential. Messages about reducing HIV sexual risk are designed to fulfill familiar relationship needs. Because trust, sexual pressure and power form the nexus of sex scripts associated with HIV sexual risk, these themes are examined through the pursuits of the various characters as sub-plots. A randomized controlled trial (RCT) with 250 high-risk women aged 18 to 29, recruited in neighborhood settings in Newark and Jersey City, will be conducted to test the effect of the video-based intervention on HIV sexual risk behavior. A control group will receive text messages with audio that stress HIV health promotion messages. The series of 12 20-minute videos to the video intervention group (n=125) and 12- weekly HIV-risk reduction text/ audio messages to the control group (n=125) will be viewed over video-capable cell phones so that women may view the videos and text messages repeatedly and in privacy. It is expected that as a result of watching the video series, the video intervention group will demonstrate greater reductions in unprotected sex from baseline to 3 and 6 months than will the control group. Further, it is expected that they will demonstrate lower sexual pressure, indicating lower stereotypical expectations to engage in unprotected sex. Secondary analyses will determine whether there are differences in these results by partner type, whether there is a reduction in the number of high risk sex partners, and whether choosing to watch the videos more frequently will relate to further reduction in risk behavior. Interviews will be conducted by audio computer assisted self-interview (ACASI). The program will track when, how often, and for how long the videos are viewed. If the aims of this proposal are achieved, the use of the cell phone to view videos could change the paradigm of how health promotion is amplified between clinic sessions. Potentially hard to reach young urban women who may be at risk for HIV, will be able to access video-based interventions on their own cell phones. The popularity of the cell phone and use of the Internet for multimedia offer a new communication channel to address health disparities in young urban women. PUBLIC HEALTH RELEVANCE: Relevance to Public Health According to the CDC, the majority (80%) of women with HIV became infected by unprotected sex with an infected male partner. Black women and Latinas accounted for 82% of AIDS diagnoses in women in 2005. Toni's Story is a series of 12 urban soap opera videos that communicates messages about reducing HIV sexual risk. By grounding the stories in women's relationship experiences, via a popular medium of the soap opera, women can identify with the heroines' emotionally charged process of change. The videos will be streamed to video-capable cell phones so that they can be viewed repeatedly and in privacy. If the aims of this proposal are achieved, potentially hard to reach young urban women who may be at risk for HIV will be able to access health promotion videos on their own cell phones. The popularity of the cell phone and use of the Internet for multimedia provide a new communication channel to address health disparities.

NATIONAL INSTITUTE OF NURSING RESEARCH Grant Number: 1R01NR010860-01A1

Thesaurus Terms:

Rutgers, The State University of New Jersey

Epigenetics of alcohol effects on stress axis development

PI: SARKAR, DIPAK sarkar@aesop.rutgers.edu

9/30/2008

Abstract: Children and young adults who were exposed to alcohol during fetal life show emotional disturbances such as depression, anxiety, attention deficit, and hyperactivity. Animal studies have linked the behavioral abnormalities to problems in the stress axis function, particularly the hyperresponsiveness of corticotrophin-releasing hormone (CRH) neuronal activity, to a variety of stressful events. Adaptation to a stressful event depends in part on an individual's ability to produce increased levels of the hormones of the stress axis and to reduce the levels of these hormones once the stressor has subsided. Recent reports indicated that hormone and toxicant exposure at crucial developmental stages results in alteration of key genes, resulting in physiological and/or behavioral changes not only in exposed individuals but also in their offspring. Hence, the question is raised whether epigenetic alterations of the CRH neuronal function are caused by alcohol exposure during the development. We hypothesize that alcohol exposure during development impairs stress axis function by altering DNA methylation and expression of proopiomelanocortin (POMC) gene in beta-endorphin neurons, which regulate CRH secretion from the hypothalamus. The objective of the proposal is to test the hypothesis that alcohol exposure during brain development produces epigenetic transgenerational effect on the stress axis function by altering DNA methylation and mRNA expression of CRH and/or its regulatory POMC genes in the hypothalamus. This will be achieved by identifying whether the stress axis hyperresponsiveness is accompanied by altered DNA methylation of gene-promoter activities in beta-endorphin and/or CRH neurons, and by evaluating whether gene-silencing activity is accompanied by the alteration in the expression of DNA methyltransferases in these neurons in the hypothalamus of offspring exposed to alcohol during the developmental period. Furthermore, whether fetal alcohol exposure induces transgenerational effects on the stress axis function will be studied. The proposed studies should provide a better understanding of the molecular mechanisms responsible for the detrimental effects of alcohol on the development of the neuroendocrine axis of stress and should reveal new putative epigenetic disease biomarkers that may enhance early detection strategies and serve as therapeutic targets for stress axis dysfunction in fetal alcohol exposed patients. PUBLIC HEALTH RELEVANCE: Recent reports have indicated that hormone and toxicant exposure at crucial developmental stages cause an alteration in the function of key genes, resulting in physiological and/or behavioral changes not only in exposed individuals but also in their offspring. Hence, the question is raised whether an inheritance of epigenetic alteration of the corticotrophin releasing hormone neuronal function induced by alcohol exposure during development causing the stress axis dysfunction that leads to various affective disorders in fetal alcohol exposed patients. The goal of this proposal is to determine the epigenetic transgenerational effect of alcohol on the stress axis function in order to identify new putative epigenetic disease biomarkers as well as to develop early detection strategies and therapeutic targets for stress axis disorders in fetal alcohol exposed patients.

NATIONAL INSTITUTE ON ALCOHOL ABUSE AND ALCOHOLISM Grant Number: 1R21AA016695-01A2

Thesaurus Terms:

ANIMAL SCIENCES

Rutgers, The State University of New Jersey