STRUCTURAL INSIGHT INTO NOVEL MECHANISMS OF TYPE III SECRETION

Start Date: 12/1/2011

Award Number: 1R01AI094623-01A1

NIH Funding Organization: NIAID

Principal Investigator: KALODIMOS, CHARALAMPOS

Co-PI: Not Applicable

FY Total Cost by IC: $141290

Program(s): Adopted;Animals;Anti-Infective Agents;ATP phosphohydrolase;Bacteria;base;Binding (Molecular Function);Biochemical;Biological Assay;biological systems;Bordetella;Cell physiology;Cell surface;Cells;Child Mortality;Complex;Cytoplasm;Cytosol;Data;Defect;design;Disease;Dysentery;enteropathogenic Escherichia coli;Eukaryotic Cell;Event;Food Poisoning;Human;In Vitro;in vivo;Infantile Diarrhea;Infection;Insecta;insight;Kinetics;kinetosome;Membrane;mimicry;Molecular Chaperones;Molecular Conformation;nanomachine;Needles;Nematoda;novel;Nucleotides;operation;pathogen;Pathogenesis;pathogenic bacteria;pathogenic Escherichia coli;Plague;Plants;Preparation;prevent;Process;Property;Protein Export Pathway;Proteins;Proton-Motive Force;prototype;Pseudomonas;Resolution;Role;Salmonella;secretion process;Shigella;Signal Transduction;Spottings;Structure;success;System;therapeutic protein;Thermodynamics;Type III Secretion System Pathway;Typhoid Fever;Vaccines;Virulence;Virulence Factors;

Abstract: DESCRIPTION (provided by applicant): A large number of bacterial pathogens, including Shigella, Salmonella, Bordetella, Pseudomonas, and pathogenic E. coli that are pathogenic for humans, other animals including insects or nematodes, and plants are equipped with a special protein- export apparatus called a type III secretion system (TTSS) or an injectisome. The injectisome is a highly sophisticated nanomachine that has specifically evolved to allow bacteria to deliver proteins into eukaryotic cells. The TTSS enables these pathogens to inject virulence proteins (known as effectors) directly into the cytoplasm of the eukaryotic host cells they infect. Many of these type III translocated effectors mimic eukaryotic factors and are capable of subverting key host cellular processes to the benefit of the pathogen during infection. Over the past decade, significant progress has been made in understanding the structure, assembly and the mode of operation of TTSS. The cytosolic components, the principal structural building proteins of the injectisome, from the basal body embedded in the inner and outer bacterial membrane to the tip of the needle protruding from the cell surface, have been extensively characterized. Virulence factors (effectors, needle proteins and translocators) form tight complexes with cognate chaperones in the cytosol and are subsequently targeted specifically to an ATPase protein located at the base of the injectisome. Powered by ATP, the effector is then translocated through the needle and is secreted in the eukaryotic cell. Fundamental questions about the functional mechanisms underpinning these processes remain unaddressed. We propose to use an integrated approach combining structural, dynamic, thermodynamic, kinetic, biochemical and in vitro and in vivo functional assays to provide insight into the early events of the secretion process that involve the recognition and binding of virulence factors (effectors and translocators) by cognate chaperones and the targeting of these substrates to the ATPase. We have extensively characterized over the last 3 years TTS protein components from the enteropathogenic Escherichia coli (EPEC), a prototype for TTSS and the major cause of infantile diarrhea and child mortality worldwide. The specific aims are designed to provide atomic-resolution insight into (i) the mechanisms of specific interaction between TTS chaperones and virulence factors, (ii) the structural and dynamic properties of the ATPase, (iii) the 'recognition' or 'secretion' signal, and (iv) the specific targeting of chaperone-substrate complexes to the ATPase. PUBLIC HEALTH RELEVANCE: Type III secretion system (TTSS) is encoded by many bacterial pathogens, including Shigella, Salmonella, Bordetella, Pseudomonas, and pathogenic E. coli that are pathogenic for humans and cause diseases such as plague, typhoid fever, bacterial dysentery and common food poisoning.

NATIONAL EPIDEMIOLOGIC SURVEY ON ALCOHOL AND RELATED CONDITIONS: DNA REPOSITORY

Start Date: 12/1/2011

Award Number: 1U24AA021085-01

NIH Funding Organization: NIAAA

Principal Investigator: TISCHFIELD, JAY ARNOLD

Co-PI: Not Applicable

FY Total Cost by IC: $3008117

Program(s): Accidents;alcohol use disorder;Alcoholism;Alcohols;Aliquot;Automation;Bacterial DNA;base;Biocompatible Materials;Biological Assay;Biological Preservation;Caring;Cells;Chemistry;Collection;cost;cost effective;design;Disease;DNA;DNA Sequence;Dyes;Engineering;Ensure;Environment;Epidemiology;Equipment and supply inventories;Ethnicity aspects;Event;Fingerprint;Future;Gender;gene environment interaction;genetic epidemiology;genetic risk factor;Genome;Genomics;Human;instrument;Laboratories;Libraries;Liquid substance;Management Information Systems;Measurement;Medical;Molecular Genetics;Molecular Weight;Monitor;National Institute on Alcohol Abuse and Alcoholism;next generation;novel;Nucleic Acids;Population;Prevention strategy;Process;Productivity;programs;Qualifying;Quality Control;repository;response;RNA;Robot;Saliva;Sampling;Secure;Sequence Analysis;Services;social;success;Surveys;System;Technology;treatment strategy;Tube;Universities;Violence;

Abstract: DESCRIPTION (provided by applicant): The National Epidemiological Surveys on Alcohol and Related Conditions (NESARC I and II) are the largest and most ambitious epidemiological surveys on alcoholism and related disorders in the civilian, non-institutionalized US population. NESARC-III is even more ambitious as it aims to collect saliva samples from as many as 46,500 subjects and extract DNA from each in an effort to identify genetic risk factors for alcohol use disorders (AUDs) and co-morbid disorders through large-scale DNA re-sequencing and to examine gene-environment interactions. We propose to establish the Saliva DNA Repository for the NIAAA NESARC-III study. Our Aims are: 1. Genomic DNA Extraction from Human Saliva Samples: We will employ state-of-the-art automation and bead-based chemistry to enrich for high molecular weight human genomic DNA relatively free of contaminants and of RNA and undesirable DNA species (i.e., bacterial DNA contamination). 2. Comprehensive Nucleic Acid Quality Control: We have engineered and validated a novel, comprehensive analytical and functional quality control (QC) program that ensures the highest quality of DNA and its utility for future sequencing analyses. Analytical QC consists of absorbance and dye-based concentration and purity measurements, precision non-contact volume measurements, and specific quantification of the amount of human genomic DNA in each sample. Functional QC employs a high-throughput, molecular genetic fingerprinting SNP assay (the RUID). We will also use several different Next Generation sequencing platforms to demonstrate the utility of this saliva DNA for future sequencing. 3. Integrated Nucleic Acid Storage and Distribution: All NESARC-III samples will be stored at -80 degrees C in dedicated freezers that are monitored 24/7/365 in a secure facility that contains state-of-the-art redundant power and HVAC systems. All sample extraction, QC, storage, biomaterial requests and distribution workflows are documented and managed by the RUCDR STARLIMS, an enterprise-level laboratory information management system. PUBLIC HEALTH RELEVANCE: Alcohol use disorders (AUDs) cost our nation billions of dollars due to lost productivity, provision of medical care, accidents, violence and the breakup of social order. In addition, they are responsible for much human misery and suffering. We must understand the causes, onset and lifetime progression of AUDs through epidemiology and genetics studies if we are to develop better treatment and prevention strategies.

EFFECT OF TISSUE TROPISM ON RNA VIRAL EVOLUTIONARY RATES: THREE FAMILY-LEVEL ANAL

Start Date: 9/30/2011

Award Number: 1R03AI096265-01

NIH Funding Organization: NIAID

Principal Investigator: DUFFY, SIOBAIN MARIE DEIRDRE

Co-PI: Not Applicable

FY Total Cost by IC: $77083

Program(s): Affect;Antiviral Agents;Antiviral Therapy;Anus;Apple;Bioinformatics;Caliciviridae;Calicivirus;Capsid Proteins;Caring;Computer software;Computing Methodologies;Country;Data;Data Set;Development;Drug resistance;Ensure;Enteral;Evolution;Family;Family Picornaviridae;foodborne illness;Gastrointestinal tract structure;Genbank;Genes;Health;Human;Human poliovirus;Immune response;Individual;Infantile Diarrhea;Intestines;member;Meta-Analysis;Methodology;Methods;Mutate;Norovirus;Nucleotides;Paper;pathogen;Pattern;Phylogenetic Analysis;Polioviruses;programs;Publishing;Relative (related person);Reoviridae;Reovirus sp.;Research;RNA Viruses;Rotavirus;Sampling Biases;Serotyping;Testing;Time;tissue tropism;Tissues;trait;treatment planning;Tropism;Vaccines;Variant;Viral;viral RNA;virology;Virus;Work;

Abstract: DESCRIPTION (provided by applicant): RNA viruses mutate and evolve more quickly than all other human pathogens. However, RNA viruses do not all evolve at the same fast rate. It is important to distinguish the faster evolving RNA viruses because they are more likely to create drug-resistant variants or evade vaccine-induced immune responses. Whole families of RNA viruses are known to be more evolvable than others, but we know very little about what causes individual viruses to evolve more rapidly than closely related viruses in the same family. We will test whether tissue tropism could be one of the factors affecting evolutionary rates. Specifically, we are testing if genes within enteric viruses, those that infect the gastro-intestinal tract, evolve more quickly than related viruses that infect other tissues in the same hosts. Long-term nucleotide substitution rates will be calculated from members of three important RNA virus families that have large numbers of sequences in GenBank: Caliciviridae, which include the leading cause of foodborne illness in this country, Picornaviridae, which include poliovirus, and Reoviridae, which include rotavirus, the most common cause of infant diarrhea worldwide. We will contrast the rates of evolution of the same gene in primarily enteric and primarily non-enteric viruses, which will allow us an apples-to-apples comparison between viruses. The BEAST software we use takes a Bayesian phylogenetic coalescent approach to calculating evolutionary rates. The accuracy of such estimates changes over different timescales, and the time to the coalescent of some of our analyses varies over an order of magnitude (i.e., some data coalesce in 70 years, others in 700 years). Since many of the enteric virus data sets coalesce more quickly than the non-enteric virus data sets, additional controls will be conducted to ensure our results are not biased by coalescent time. Our work has the potential to open up avenues of research into whether enteric tissue tropism mechanistically leads to faster rates of RNA viral evolution, and to highlight weaknesses in a widely used software program. PUBLIC HEALTH RELEVANCE: Faster evolving RNA viruses are more difficult to control and treat than slower evolving RNA viruses, but we have not identified many traits that cause viruses to evolve more quickly. This research tests whether enteric viruses, those that infect the mammalian gastrointestinal tract such as poliovirus, norovirus and rotavirus, evolve more quickly than their relatives that infect other mammalian tissues. If infecting enteric tissue leads to faster rates of viral evolution, virologists will know to take more care can when developing vaccines and durable antivirals for enteric viruses.

CENTER FOR EDUCATION AND RESEARCH ON MENTAL HEALTH THERAPEUTICS

Start Date: 9/30/2011

Award Number: 1U19HS021112-01

NIH Funding Organization: AHRQ

Principal Investigator: CRYSTAL, STEPHEN

Co-PI: Not Applicable

FY Total Cost by IC: $

Program(s):

Abstract: DESCRIPTION (provided by applicant): The Rutgers-Columbia Center for Education and Research on Mental Health Therapeutics will focus on safe and effective use of therapeutics for persons with behavioral health disorders. Center projects and activities will address critical gapsin the evidence base on the comparative effectiveness of mental health treatments and services system innovations. We will build on the solid foundation of infrastructure, partnerships, dataset development, and lines of research from our first five years of work as a CERTs Research Center to address all five programmatic interest areas. The proposed work will be guided by systematic stakeholder input through an advisory committee representing organizations that are well positioned to translate Center findings into improved care processes and outcomes. Research and educational activities will be supported by a Data, Methods and Biostatistics Core and an Education, Translation and Stakeholder Core and organized within population-based cores for 1) youth, 2) adults, and 3) the frail elderly. Specific projects include: 1) a comparativ effectiveness analysis of alternative pharmacological strategies to manage treatment-resistant schizophrenia, 2) an analysis of the impact of an innovative statewide community mental health center-based medical home model for the safe and effective use of mental health therapeutics, 3) a comparative safety analysis of anti-dementia medications in the nursing home elderly, 4) an examination of the potential impact of structured diagnostic interviewing on the management and outcome of youth with severe psychiatric disorders, and 5) an assessment of premature mortality in schizophrenia patients and their underlying quality of general medical care. Through these projects and related educational activities, the Center aims to serve as a trusted national resource to inform decision's making of mental health stakeholders, with an emphasis on broad populations -under treatment, especially in Medicaid and Medicare. Our experience working with states and federal policy makers also qualifies us to provide leadership of new cross-CERTs initiatives involving Medicaid agencies and complex, high-cost patients PUBLIC HEALTH RELEVANCE: Mental health conditions account for a substantial share of overall illness burden and health care expenditure, and substantially complicate the course and treatment of other medical disorders. The Center's work will address these critical issues, serving as a national observatory on the use and outcomes of mental health treatments in large, usual-care settings, incorporating strong partnerships with key stakeholders.

NEURON-SPECIFIC EFFECTS OF IL-1B

Start Date: 9/15/2011

Award Number: 1R21NS076867-01

NIH Funding Organization: NINDS

Principal Investigator: FRIEDMAN, WILMA J

Co-PI: Not Applicable

FY Total Cost by IC: $231250

Program(s): Astrocytes;Attenuated;Binding (Molecular Function);Brain;Brain Injuries;cell type;Cells;Complex;cytokine;Disease;Event;Goals;hippocampal pyramidal neuron;Hippocampus (Brain);Inflammation;Inflammatory;inhibitor/antagonist;Injury;Interleukin-1;interleukin-1 receptor accessory protein;Interleukin-1 Receptors;Interleukins;Long-Term Potentiation;Mediating;Microglia;Mus;Neuroglia;neuron loss;Neurons;novel;Outcome;Physiological;Play;Population;Production;Protein Isoforms;Proteins;receptor;response;RNA Splicing;Role;Signal Pathway;Signal Transduction;Signaling Protein;Stress;synaptic function;Therapeutic;therapeutic development;therapy development;tool;Variant;Wild Type Mouse;

Abstract: DESCRIPTION (provided by applicant): Inflammation occurs as a consequence of brain injury, and results in the production of inflammatory cytokines, which contribute to the progression of the insult. One of the most potent inflammatory cytokines is interleukin-1 (IL-1), which is induced by many types of brain injury and can potentiate neuronal loss. IL-1 inhibitors have been suggested as therapeutic tools following brain injury to attenuate neuronal loss. However, IL-1¿ is also expressed in the brain under physiological conditions, and may play a role in mediating normal hippocampal function. We have previously demonstrated that IL-1¿ activates distinct signaling pathways in neurons versus glial cells in the brain. Therefore, to assess the consequences of inflammation that occur following injury, and the potential therapeutic value of inhibiting inflammation, it is critical to understand the cell-specific consequences and to distinguish physiological versus pathophysiologial roles for this key cytokine. IL-1 signals via a receptor complex composed of the type 1 IL-1 receptor (IL-1R1) and the IL-1 Receptor Accessory Protein (IL-1RAcP). A novel isoform of the IL-1RAcP, termed AcPb, has been identified that is expressed exclusively in CNS neurons, providing a new opportunity to investigate the mechanisms governing neuron-specific IL- 1 actions. To identify the function of the neuron-specific IL-1RAcPb we will establish neuronal cultures from AcPb-/- mice compared to wild type neurons and to neurons lacking both isoforms of the protein (AcP-/-). The effects of IL-1¿ on these neurons will be analyzed to determine the role of AcPb in mediating neuron-specific signaling pathways, and the consequences for neuronal function. PUBLIC HEALTH RELEVANCE: Interleukin-1 is a potent and pleiotropic cytokine that is induced in the brain by injury, disease, and stress. The recent identification of a neuron-specific isoform of a key IL-1 signaling protein provides a novel opportunity to investigate mechanisms governing the specific effects of IL-1 on neuronal function. These studies may also facilitate the development of therapeutic approaches to distinguish physiological versus pathophysiologial roles for this key cytokine in the brain.

1/8-COLLABORATIVE GENOMIC STUDIES OF TOURETTE DISORDER

Start Date: 9/15/2011

Award Number: 1R01MH092293-01A1

NIH Funding Organization: NIMH

Principal Investigator: HEIMAN, GARY A.

Co-PI: TISCHFIELD, JAY ARNOLD

FY Total Cost by IC: $190874

Program(s): Affect;Alleles;Attention deficit hyperactivity disorder;base;Biocompatible Materials;Caucasians;Caucasoid Race;Cell Line;Child;Clinical;Collaborations;Collection;Communities;Comorbidity;Complementary DNA;Complex;Copy Number Polymorphism;cost;Data;data management;Development;Diagnostic;Disease;DNA;DNA Sequence;European;exome;experience;Family;Family member;follow-up;Future;gene discovery;Gene Expression;Gene Expression Profile;Genes;Genetic;genetic pedigree;Genetic Predisposition to Disease;genome wide association study;genome-wide;Genomics;Genotype;Gilles de la Tourette syndrome;Health Benefit;Heterogeneity;Human;Individual;International;Joints;Koreans;Measures;member;Mental disorders;Molecular;Motor Tics;Mutation;neuropsychiatry;next generation;novel;Obsessive-Compulsive Disorder;Parents;Pilot Projects;Population;Prevalence;proband;public health medicine (field);Recruitment Activity;Research Project Grants;Resources;Risk;RNA;Sample Size;Sampling;Site;Symptoms;Syndrome;Techniques;Time;Translating;Tube;Variant;Vocal Tics;

Abstract: DESCRIPTION (provided by applicant): SUMMARY/ABSTRACT: Tourette Disorder (TD) is a developmental neuropsychiatric syndrome characterized by the combination of persistent vocal and motor tics. While initially considered rare, the world-wide prevalence is now estimated to be 0.3-1%. Both as a consequence of potentially disabling symptoms as well as very high rates of psychiatric co-morbidity, particularly with obsessive-compulsive disorder (OCD) and attention deficit hyperactivity disorder (ADHD), TD represents a significant public health concern. Despite decades of evidence supporting a significant genetic contribution, progress in identifying risk alleles has been slow. This difficulty is thought to be, in part, a consequence of complex inheritance and substantial genetic and phenotypic heterogeneity. This collaborative study unites an international group of highly expert clinicians specializing in TD with statistical and molecular geneticists and is motivated by three central hypotheses: 1) that a key rate- limiting factor for TD gene discovery has been the paucity of publically available, large-scale biomaterial resources of the kind that are now commonplace for many neuropsychiatric disorders; 2) based on recent data from a host of other genetically complex disorders, a comprehensive genomics study of TD will require large samples sizes and should focus on the potential contribution of rare as well as common alleles and both sequence and structural variants; and 3) an increased understanding of the genetic etiology of TD will translate into novel and more effective approaches to treating this often-debilitating disorder, and consequently will have marked public health benefits. The application elaborates three specific aims: Specific Aim 1: To recruit 5050 individuals with TD (and their family members), and make DNA, cell-lines, cDNA/RNA and phenotypic data publicly available within one year of collection. The sample will include a subset of 3195 European Caucasian (EC) probands; 1250 Korean probands, and 3295 parent-child trios allowing for the study of de novo variation. We will also recruit each year at least 10 TD pedigrees with 4 or more affected members as a resource for family-based gene discovery. Specific Aim 2: To employ state-of-the-art techniques to identify and confirm rare and common variants contributing to TD. We will genotype the sample on Illumina HumanOmni2.5 -Quad BeadChips to support copy number variation (CNV) analysis (Aim 2A) and genome wide association studies (GWAS) (Aim 2B); whole exome sequencing will be employed in select, multiply-affected TD pedigrees (Aim 2C); and we will follow up on the most promising loci identified in the aforementioned studies using a pooled next generation re-sequencing strategy (Aim 2D) at two time points, evaluating a minimum of 50 genes in a total of 3195 EC and 3195 matched controls; Specific Aim 3: To perform preliminary analyses of 300 transcriptomes of TD subjects to investigate the implications of selected structural and sequence variations for cis, trans and genome-wide expression. With no cost to this project, PAXgene tubes will be collected from all subjects and made available to the scientific community to enable future studies by our group and others. PUBLIC HEALTH RELEVANCE: Project Narrative: Tourette Disorder (TD) is a poorly understood, often debilitating neuropsychiatric disorder characterized by a combination of persistent vocal and motor tics, affecting between 3 in 1000 and 1 in 100 people worldwide, predominantly children. Our international team of experts in TD and other mental disorders proposes to recruit 5050 affected individuals, undertake a comprehensive gene discovery effort, and make the collected biomaterials and clinical data rapidly available to the broad scientific community, thereby enabling both current and future TD research. Understanding the causes of TD may facilitate the development of novel and improved treatments.

PHENOTYPIC ANALYSIS OF HUMAN IPSC CARRYING ADDICTION-ASSOCIATED GENE VARIANTS

Start Date: 9/15/2011

Award Number: 1R21DA032984-01

NIH Funding Organization: NIDA

Principal Investigator: TISCHFIELD, JAY ARNOLD

Co-PI: HART, RONALD P

FY Total Cost by IC: $231000

Program(s): addiction;Addictive Behavior;Adoption;Adult;Affect;Alleles;Amino Acids;Archives;Authorization documentation;base;Behavior;Biochemical;Biological Assay;Blood;Blood specimen;Brain Diseases;Calcium;cell bank;Cell Culture Techniques;Cell Line;Cell model;Cell physiology;Cell Surface Receptors;cell type;Cells;Child Abuse;Cholinergic Receptors;Chronic;Collection;cost;Crime;Cultured Cells;Data;DNA;Drug abuse;Drug Addiction;Electrophysiology (science);embryonic stem cell;Employment;Episome;expectation;Family;Gene Expression;gene function;Genes;genetic variant;Genomics;Genotype;Goals;Health;Housing;Human;Image;immunocytochemistry;Individual;induced pluripotent stem cell;Inherited;Intervention;Laboratories;Lymphocyte;Maps;Methods;Modeling;Molecular;molecular marker;molecular modeling;Molecular Models;Morphine;Mutate;Mutation;National Institute of Drug Abuse;National Institute of Mental Health (U.S.);National Institute on Alcohol Abuse and Alcoholism;nerve stem cell;Neuronal Differentiation;Neurons;Nicotine;Nicotinic Receptors;novel;novel strategies;Opioid;Opioid Receptor;Patients;Pharmaceutical Preparations;Pharmacology;Phenotype;Physiological;Physiology;pluripotency;Property;Protocols documentation;Publishing;release of sequestered calcium ion into cytoplasm;Reporting;repository;Research;Research Personnel;Resources;response;Risk Factors;Sampling;Screening procedure;second messenger;Second Messenger Systems;Signal Transduction;Societies;Source;Staging;Stem Cell Research;Stem cells;Study models;Substance abuse problem;Substance of Abuse;System;Techniques;Technology;Testing;Time;Universities;Variant;Variation (Genetics);vector;Viral;

Abstract: DESCRIPTION (provided by applicant): Identification of genetic variations associated with addictive behaviors provides a novel opportunity for using cell cultures to model molecular and cellular mechanisms that underlie addiction. The observed genetic variations have been mapped to amino acid changes in cell surface receptors, presumably affecting neuronal circuits involved in addiction behaviors. However, cellular context is likely to be important in determining the function of these genes. Construction of induced pluripotent stem cells (iPSC) from adult cells derived from drug abusing individuals carrying known genetic variants provides a means for developing physiologically relevant culture systems for understanding addiction. For the culture cell model to be useful it must not only express the affected gene but it must also provide an appropriate cellular context for studying pharmacology or cell signaling. There is no clear expectation about what type of iPSC-derived cultures would be useful in studying the cellular physiology altered by gene variants. Our hypothesis is that the altered physiology of genetic variants associated with addiction liability can be modeled in cultured neurons derived from iPSCs constructed from donor lymphocyte samples. We propose to construct several iPSC lines from donor lymphocyte samples, differentiate these iPSC into functional neurons, and then to develop methods to assay possible phenotypic differences between variant-derived cells and wild-type. These cells will be valuable for identifying cell and molecular responses to substances of abuse, to examine the effects of a known genotype on the cellular phenotype, as well as to develop novel approaches for pharmacologic intervention. PUBLIC HEALTH RELEVANCE: Drug abuse and addiction are a major burden to society, with the total cost of substance abuse in the U.S. exceeding $500 billion annually. Addiction is a chronic brain disease that leads to a compulsive desire for drugs despite significantly harmful consequences, including direct effects on health as well as detriments to society, including disintegration of families, child abuse, loss of employment, and a broad array of crimes. While addiction may be treatable, the discovery that some forms of addiction have an inherited component demonstrates that physiology contributes to addictive behaviors. To exploit new techniques of cellular reprogramming to prepare human neuronal cultures, we will establish induced pluripotent cell (iPSC) lines derived from donor blood samples carrying mutations in two genes known to be risk factors for addiction-an acetylcholine receptor and an opioid receptor. The blood lymphocyte samples already exist in the NIDA Center for Genomic Studies collection and will be used with permission from the collecting investigator. These iPSC will then be induced to differentiate into specific types of neurons for use in investigating the molecular bases of addiction and for screening new treatment therapies.

MODELING AGE-ASSOCIATED NEURONAL SPROUTING AND FUNCTIONAL DECLINE IN A GENETIC SY

Start Date: 9/15/2011

Award Number: 1R21AG041498-01

NIH Funding Organization: NIA

Principal Investigator: DRISCOLL, MONICA A.

Co-PI: Not Applicable

FY Total Cost by IC: $169494

Program(s): Address;Adult;Age;Aging;aging brain;Aging-Related Process;Alzheimer's Disease;Animals;Appearance;Axon;base;Biology;biomarker;Brain;Caenorhabditis elegans;cell body (neuron);Cell Death;cell type;Cells;Cessation of life;combat;Development;dietary restriction;Disease;Documentation;drug discovery;drug efficacy;drug use screening;Environmental Risk Factor;Exhibits;Experimental Models;Foundations;Frequencies (time pattern);functional decline;Future;Genes;Genetic;genetic analysis;genetic technology;Germ Cells;Health;healthy aging;Human;Imaging technology;Impaired cognition;improved;Individual;insight;insulin signaling;interest;Intervention;Investigation;Label;Lead;Life;Longevity;Maintenance;Metformin;Microscope;mimetics;Mitochondria;mitochondrial dysfunction;Modeling;Molecular;Monitor;Nematoda;Nerve Degeneration;Nervous system structure;Neurobiology;neuron loss;Neuronal Dysfunction;Neurons;neuroprotection;new growth;novel;Pathway interactions;Pharmaceutical Preparations;Phenotype;Physiological;Predisposition;Process;protein aggregation;receptor;Reporter;research study;Resveratrol;Risk Factors;Structure;Surveys;Synapses;synaptic function;system architecture;Testing;Therapeutic Intervention;Tissues;Touch sensation;Work;

Abstract: DESCRIPTION (provided by applicant): Subtle changes in the aging human brain, such as neuronal sprouting and restructuring, are thought to underlie cognitive decline and may promote neuronal dysfunction in Alzheimer's disease. We study fundamental processes relevant to neuronal function and neurodegeneration in the 959-celled nematode C. elegans. In this transparent roundworm, we can directly observe individual fluorescently labeled neurons, as well as synapses and mitochondria, in the living animal. Recently we carefully documented how some C. elegans neurons change as animals grow old. We find that like human brain aging, there is little neuron loss as C. elegans ages. However, some types of neurons exhibit dramatic novel branching and outgrowth with age. We can score new growth using a high magnification dissecting microscope, which facilitates relatively rapid analysis within live, aging animals. We propose to characterize morphological neuronal aging in C. elegans to establish a facile model for analysis of genetic and environmental factors that influence neuronal aging. Aim 1 is to extend our initial cell-specific analysis to diverse neuronal classes to better define what neuronal types are susceptible to age-associated morphological changes. This tractable work will document the basic neurobiology of C. elegans aging, definitively establishing an overview of differential susceptibility of specific neurons to age-associated morphological change. Aim 2 is to survey four mechanisms hypothesized to contribute to age-associated neuronal regrowth (neuronal function, synaptic function, disrupted proteostasis, and mitochondrial changes) to identify which contributes to morphological aging. In this aim we might also identify fluorescent synaptic and mitochondrial indicators of neuronal aging. These experiments may suggest the most potent factors in age-associated neuronal restructuring. Aim 3 is to test whether dietary restriction, a conserved mechanism for lifespan extension that has been found to improve human neuronal aging, can protect against neuronal morphological aging phenotypes. We will also test DR-mimetic drugs for efficacy in the nematode model. This work may identify a conserved mechanism for neuroprotection and establish proof-of-principle for the suitability of this new model to be used in for anti-neuronal aging drug discovery. Our work will define the first model for high throughput genetic and drug studies of age-associated neuronal morphological abnormalities that may inform on mechanisms and interventions that can better maintain the aging human nervous system. PUBLIC HEALTH RELEVANCE: Neuronal aging in the human brain, a major risk factor for Alzheimer's disease also proposed to influence cognitive decline in healthy aging, is associated with synaptic changes and aberrant sprouting. Recently, we discovered that some neurons in aging C. elegans exhibit aberrant branching and outgrowth and we have documented a global decline of synaptic structure with age, suggesting that fundamental neuronal aging mechanisms may be conserved from simple animals to humans. We plan to develop the C. elegans model of neuronal aging to describe susceptible neurons and identify process that can promote or protect against neuronal decline. Documentation of this model should lead to advanced mechanistic understanding that might ultimately suggest novel and efficacious therapies to combat human brain aging.

PTSD AND ADDICTION DISORDER SCREENING AND TREATMENT FOR INCARCERATED MEN

Start Date: 9/2/2011

Award Number: 1R01MH095206-01

NIH Funding Organization: NIMH

Principal Investigator: WOLFF, NANCY

Co-PI: Not Applicable

FY Total Cost by IC: $731468

Program(s): addiction;Address;Administrator;Adult;aged;Alcohol consumption;Alcohol or Other Drugs use;arm;Attention;base;Behavioral;behavioral health;care systems;Cognitive;compare effectiveness;Computer software;computerized;Computers;cost effective;Development;Diffuse;Disease;Drops;Drug usage;effective intervention;Effectiveness;emotional abuse;empowerment;evidence base;Evidence based intervention;experience;Focus Groups;Freedom;Goals;group intervention;Health;Health Personnel;Housing;Imprisonment;improved;Individual;informant;Institute of Medicine (U.S.);Intervention;Interview;Lead;Left;Life;male;medical schools;men;Mental disorders;Mental Health;Methods;mindfulness;mindfulness meditation;Modality;Modeling;National Institute of Mental Health (U.S.);New Jersey;Outcome;Phase;phase 1 study;physical abuse;Population;population based;Post-Traumatic Stress Disorders;preference;Prisons;provider intervention;public health medicine (field);Randomized;Randomized Controlled Trials;Recovery;Reporting;Research Support;Risk Factors;Safety;Screening procedure;Security;Services;Sexual abuse;skills;Staging;Strategic Planning;Substance Use Disorder;Surgeon;Testing;Time;Trauma;Trauma recovery;treatment as usual;Veterans;Work;

Abstract: DESCRIPTION (provided by applicant): At least one-fifth of men inside state prisons have mental health problems, over half have experienced physical, sexual, or emotional abuse (referred to as 'interpersonal trauma') during their formative years and often in adulthood, and the majority has alcohol or drug use problems. Interpersonal trauma is highly correlated with health and behavioral health problems and criminality. Few male inmates receive treatment for behavioral health disorders related to interpersonal trauma while incarcerated even though these problems are risk factors for returning to prison after release. While the Surgeon General's Report and the President's New Freedom Commission Report highlighted the importance of trauma-informed services and systems of care, trauma- related behavioral health problems among incarcerated men have received little attention. This application addresses both screening for and treatment of Post-Traumatic Stress Disorder (PTSD) and substance use disorders (SUD) within an adult male correctional setting. The proposed study will be conducted at an adult medium security prison in New Jersey that houses ~1900 male inmates primarily aged 18 to 25. We propose a two-phase study that: (1) tests the reliability (test-retest) of computer-administered screening for PTSD and SUD compared to clinician-administered screening (goal: to develop a cost-effective population- based screening procedure for PTSD and SUD within incarcerated settings) and (2) conduct a randomized controlled trial (RCT) in which 240 male inmates are assigned to two evidence-based, manualized first stage trauma interventions (Seeking Safety (SS) and Trauma Recovery and Empowerment Model version for men with substance use problems (M-TREM) (goal: to identify effective interventions for PTSD and SUD among incarcerated men). The RCT will be enhanced in two ways: (1) those who decline treatment participation will be shown a 10-minute educational video on PTSD and SUD and after viewing the video invited a second time to participate in the RCT and (2) determined by random assignment, two-thirds of participating subjects (n=160) will be randomly assigned to treatment and the other one-third (n=80) will choose treatment based on their preference for one intervention over the other. We will be testing overall effectiveness of screening and treatment interventions and the marginal impact of educational-enhancement and preferences on outcomes. PUBLIC HEALTH RELEVANCE: This study has the potential to identify cost-effective methods for screening PTSD and SUD among incarcerated men and effective interventions for treating these conjoint behavioral health problems. The importance of focusing on PTSD screening and trauma-informed treatment was highlighted in the Surgeon General's Report, the President's New Freedom Commission on Mental Health and a variety of reports by The Institute of Medicine. This application is responsive to the current NIMH Strategic Plan, specifically Objectives #3 (Develop New and Better Interventions that Incorporate the Diverse Needs and Circumstances of People with Mental Illnesses) and #4 (Strengthen the Public Health Impact of NIMH-Supported Research).

AN INTEGRATED PROBABILISTIC FRAMEWORK FOR SHAPE AND SURFACE INTERPRETATION

Start Date: 9/1/2011

Award Number: 1R01EY021494-01A1

NIH Funding Organization: NEI

Principal Investigator: FELDMAN, JACOB

Co-PI: SINGH, MANISH

FY Total Cost by IC: $294099

Program(s): Accounting;Architecture;base;Brain;Classification;Code;Cognitive deficits;Collection;Comprehension;Computer Simulation;Conceptions;Data;Disease;Figs - dietary;Functional disorder;Goals;Growth;Human;human subject;Image;Literature;Medial;Methods;Modeling;Nervous system structure;neural model;novel;novel strategies;object recognition;Ownership;Perception;Plague;programs;Psychological Models;reconstruction;Research;research study;Shapes;skeletal;Skeleton;Source;Stochastic Processes;Structure;Surface;Techniques;Testing;Texture;theories;Universities;Visual;Visual Cortex;Visual system structure;Work;

Abstract: DESCRIPTION (provided by applicant): An integrated probabilistic framework for shape and surface interpretation Jacob Feldman and Manish Singh, Rutgers University The representation of visual shape is one of the central problems of perception, influencing many aspects of object recognition and scene understanding. But a comprehensive and principled account of how the human brain computes shape representationsdoes not yet exists. A key difficulty is in understanding how the brain divides the image into distinct surfaces at distinct depths, interprets the 3D shape of each surface, and divides each shape into distinct parts. In previous work the PIs have developed novel, principled mathematical methods for understanding human shape representation, based around the idea of Bayesian estimation of the shape skeleton. The shape skeleton is a representation of the axial structure of the shape, related to though different from classical medial axis representations. Medial axis representations break shapes down into their component axes, about which the shape is approximately locally symmetric. The PI's approach recasts this as a probabilistic inference problem, consistent with most contemporary neurocompuational modeling, but unlike most other computational models of shape. This allows the approach to be expanded in scope, encompassing the broader problem of the decomposition of the image into distinct surfaces and the interpretation of 3D shape. The approach is theoretically unified, and is mathematically suitable to be implemented in a parallel computational architecture, making it plausible as a model of neural shape coding. The aim of this research program is to fully develop shape representation as a probabilistic es- timation problem, and test the many empirical predictions that emanate from this framework. Specific aims include expanding and testing the probabilistic approach to shape representation, and generalizing it to critical related problems, including figure/ground interpretation and 3D shape. This expansion will make it possible to fully integrate shape representation with the comprehension of surfaces in the visual image, in a coherent mathematical framework consistent with what is known about computation in visual cortex. PUBLIC HEALTH RELEVANCE: This project involves empirical studies of critical functions of the human visual system, including the interpretation of visual shapes and surfaces. The research contributes to a foundational understanding of the human nervous system, thus helping us achieve better functional understanding of the mechanisms of disease in perceptual dysfunctions and cognitive deficits.

ESTROGEN REGULATION OF CHANNELS INVOLVED IN THE CONTROL OF ENERGY HOMEOSTASIS

Start Date: 9/1/2011

Award Number: 4R00DK083457-03

NIH Funding Organization: NIDDK

Principal Investigator: ROEPKE, TROY ADAM

Co-PI: Not Applicable

FY Total Cost by IC: $248287

Program(s): Acute;Address;Adult;Affect;Agonist;Animals;Attenuated;bisphenol A;Cations;Cell physiology;cell type;Cells;Coupled;Development;Disease;Electrophysiology (science);Endocrinology;Environmental Estrogen;Epidemic;Estradiol;Estrogen Receptor alpha;Estrogen Receptors;Estrogens;Etiology;Event;Exposure to;Female;G Protein-Coupled Receptor Genes;G-Protein-Coupled Receptors;Gene Expression;Gene Expression Regulation;Goals;Health;Homeostasis;Hormone replacement therapy;Human;Hypothalamic structure;Instruction;Knowledge;Lead;Learning;Ligands;Link;Measures;Mediating;Membrane;Membrane Potentials;Mentors;mouse model;Mouse Strains;Neurologic;neuronal excitability;Neurons;Neurosciences;novel;novel strategies;Obesity;Oils;Ovariectomy;Patch-Clamp Techniques;Pathway interactions;Phase;Physiological;Play;Polychlorinated Biphenyls;Postmenopause;Property;Regulation;reproductive;Research;Role;Signal Transduction;Steroids;Techniques;Time;Transgenic Mice;Transgenic Organisms;Weight Gain;Wild Type Mouse;Woman;Women's Health;

Abstract: No abstract available.

BIOMEDICAL (APPLIED/EXPLORATORY)

Start Date: 9/1/2011

Award Number: 276201100016C-0-0-1

NIH Funding Organization: NLM

Principal Investigator: TISCHFIELD, JAY

Co-PI: Not Applicable

FY Total Cost by IC: $1435419

Program(s): Blood specimen;Cell Line;Clinical;Collection;Communities;computerized data processing;Contracts;cost effective;Data;Disease;DNA;Ensure;Ethics;Funding;Genes;Genetic;Genetic Determinism;Genetic Materials;genetic pedigree;genetic variant;Grant;immortalized cell;Individual;Institutes;Lymphocyte;Maintenance;Mission;National Institute of Diabetes and Digestive and Kidney Diseases;Participant;Population;Population Control;Qualifying;Quality Control;repository;Research;Research Personnel;Resource Sharing;Resources;RNA;Sample Size;Sampling;United States National Institutes of Health;Work;

Abstract: Discovery of disease related genes requires a population of individuals with the genetic variant, as well as a population of control (unaffected) individuals. Thus, a repository of genetic materials, including DNA samples, cryopreserved lymphocytes, immortalized cell lines, and accompanying clinical and pedigree data is clearly an invaluable resource for the research community studying diseases related to the mission of the NIDDK. This project, which will be referred to herein as ¿The Genetics Repository¿, continues an important collection begun in 2003 and is conceptually related to repositories in existence at other institutes at the NIH. The repository will allow the continued storage, maintenance, and quality control, and equitable, ethical distribution of DNA and other resources important to the study of diseases under the mandate of the NIDDK. It facilitates sharing of resources, thus encouraging work by a broad group of investigators and, perhaps, increasing the sample size and the resulting power of a study to identify genetic determinants of a disease. It helps to ensure that research participants will be making a maximal contribution, and decreases duplicative sampling efforts.

NECTIN-LIKE CELL ADHESION MOLECULES AND THE MECHANISMS OF MYELINATION

Start Date: 9/1/2011

Award Number: 1R01NS065218-01A2

NIH Funding Organization: NINDS

Principal Investigator: MAUREL, PATRICE

Co-PI: Not Applicable

FY Total Cost by IC: $337,240

Program(s): Address;Adhesions;Axon;Binding (Molecular Function);Cell Adhesion;Cell Adhesion Molecules;Cell Differentiation process;Characteristics;Complex;Data;Development;Electron Microscopy;Exhibits;Extracellular Domain;Fiber;Goals;Image;In Vitro;in vivo;insight;knock-down;loss of function;Mediating;Mediator of activation protein;Membrane;Molecular;mutant;Myelin;myelination;nectin;Neuregulin 1;Neuroglia;Neurons;Neuropathy;NGFR Protein;novel;novel therapeutics;PAWR gene;Peripheral;Peripheral Nervous System;Phenotype;Proteins;receptor;Receptor Protein-Tyrosine Kinases;Receptor Signaling;Recruitment Activity;Reporting;research study;Resolution;Role;scaffold;Schwann Cells;Signal Transduction;small hairpin RNA;Staging;Testing;transcription factor;

Abstract: DESCRIPTION (provided by applicant): Axon-glial interactions are critical for the induction of myelination and the domain organization of myelinated fibers. We recently reported that in the peripheral nervous system (PNS) neurons and myelinating Schwann cells express a new set of cell adhesion molecules called the Nectin-like (Necl) proteins. In particular, Necl-1 on axons mediates adhesion by specifically binding to Necl-4 on Schwann cells. We showed that Necl-4 is required to initiate PNS myelination. These results implicate the Necl proteins as new, crucial mediators of axon-glia interactions. A key question is how the Necl proteins, and in particular Necl-4, mediates Schwann cell myelination. The Necls mediate cell adhesion though their Ig-like extracellular domain, and interact with scaffolding intracellular proteins through a FERM- and PDZ-binding domains. Through these latter interactions the Necl proteins can assemble polarity and signaling complexes. In preliminary studies we have obtained evidence that Necl-4 interacts with the polarity protein Par-3. These studies have also pointed out a possible interaction (direct or indirect) between Necl-4 and the Schwann cell ErbB receptors that transduce the instructive, axon-derived myelinating signal. Using mutant versions of Necl-4 in combination with lentiviral knockdown/rescue strategies, in vivo an in vitro, we will characterize the requirement of the PDZ-binding domain of Necl-4 in the initial stages of Schwann cell myelination, address the functional significance of the Necl-4/Par-3 interaction, and investigate how Necl-4 influence ErbB receptor signaling. The overall goal of this project is to provide novel insights into the mechanisms that regulate myelin formation in the PNS. A detailed understanding of the molecular mechanism by which components, required for myelination, cooperates is highly significant, and should provide valuable insights for the development of new therapeutic strategies to promote remyelination in peripheral demyelinating neuropathies. PUBLIC HEALTH RELEVANCE: The overall goal of this project is to provide novel insights into the mechanisms that regulate axon- glia interactions and myelin formation in the PNS. A detailed understanding of the molecular mechanism by which components, required for myelination, cooperates is highly significant, and should provide valuable insights for the development of new therapeutic strategies to promote remyelination in peripheral demyelinating neuropathies.

DISPOSITION OF ENVIRONMENTAL CHEMICALS DURING PREGNANCY

Start Date: 9/1/2011

Award Number: 1R01ES020522-01

NIH Funding Organization: NIEHS

Principal Investigator: ALEKSUNES, LAUREN M

Co-PI: Not Applicable

FY Total Cost by IC: $493634

Program(s): Adverse effects;ATP phosphohydrolase;Back;Biological Assay;Blood Circulation;Cells;Chemical Exposure;Chemicals;Communities;Data;design;Development;Diabetes Mellitus;Endocrine Disruptors;Endocrine system;Environment;environmental chemical;Epigenetic Process;epigenetic variation;Exposure to;fetal;Fetal Tissues;Fetus;Genetic;Genetic Polymorphism;genetic variant;Genistein;Goals;Human;human ABCG2 protein;in utero;In Vitro;in vivo;Knowledge;Laboratories;loss of function;Malignant Neoplasms;mammary gland development;Membrane;MicroRNAs;Modeling;Mus;Mycotoxins;Obesity;offspring;overexpression;Patients;Perinatal Exposure;Pesticides;Phytoestrogens;Placenta;placental transfer;Plasticizers;Poisons;Precocious Puberty;Predisposition;Pregnancy;Pregnant Women;prenatal;prenatal exposure;prevent;protein expression;protein function;protein transport;Regulation;reproductive;reproductive development;Reproductive system;Research;response;Risk;Rodent;Role;Source;Staging;Testing;Tissues;Toxic Environmental Substances;toxicant;translational study;Untranslated RNA;Variant;Variation (Genetics);Vesicle;Voice;Xenobiotics;

Abstract: DESCRIPTION (provided by applicant): Over the last decade, there has been increasing concern regarding disruption of endocrine and reproductive systems as a consequence of in utero exposure to xenobiotics. Key to preventing these responses is identification of cellular mechanisms that regulate prenatal disposition of endocrine disrupting chemicals (EDCs). One potential way to reduce levels of EDCs in the fetus is to transport them across the placenta back to the maternal circulation. One prominent transporter, breast cancer resistance protein (BCRP), has been shown to participate in the placental transfer of chemicals. Recent data from this laboratory demonstrate that the model EDC, the phytoestrogen genistein, is a BCRP substrate, and that various EDCs inhibit BCRP transport. We hypothesize that BCRP maintains low concentrations of EDCs in the fetal compartment by actively transporting them back to the maternal circulation. This is important because impaired BCRP function would increase fetal levels of EDCs and enhance disruption of reproductive development. This hypothesis will be tested using a combination of in vitro and in vivo approaches that evaluate BCRP transport of EDCs and identify the regulatory mechanisms that influence BCRP expression and activity in human placenta. We anticipate that EDCs will be BCRP substrates and that reduced Bcrp transporter function will increase EDC levels in mouse fetuses. Further, it is expected that Bcrp-null offspring will have more profound abnormalities in reproductive and mammary gland development following prenatal genistein exposure, compared to wild-types. These studies will fill significant knowledge gaps in the fields of placental transport and fetal susceptibility to environmental toxicants. The proposed research is critically needed because multiple sources of EDCs exist in our environment and exposure to EDCs has been documented in pregnant women. A detailed understanding of the regulation of placental BCRP will allow us to predict which patients are at greater risk for adverse effects of EDCs. PUBLIC HEALTH RELEVANCE: The mechanism by which the placenta removes toxic chemicals away from the fetus is poorly understood. These studies will determine whether the placental transporter BCRP reduces in utero exposure to endocrine disrupting chemicals and protects the fetus from developmental abnormalities.

POLYCYSTINS AND CILIA IN C. ELEGANS

Start Date: 9/1/2011

Award Number: 2R01DK059418-12

NIH Funding Organization: NIDDK

Principal Investigator: BARR, MAUREEN M

Co-PI: Not Applicable

FY Total Cost by IC: $372639

Program(s): 1-Phosphatidylinositol 4-Kinase;Address;Affect;Afferent Neurons;Animals;Autosomal Dominant Polycystic Kidney;Behavior;Biochemical;Biological;Biological Models;Biology;Caenorhabditis elegans;Candidate Disease Gene;Cells;Cellular biology;Chromosome Mapping;Cilia;ciliopathy;cilium biogenesis;Complex;Cystic kidney;Defect;Development;Diffusion;Disease;Double-Stranded RNA;Down-Regulation;Electrophysiology (science);End stage renal failure;Enzymes;Essential Genes;Esthesia;Excision;feeding;Genes;Genetic;Goals;Health;Hereditary Disease;Homologous Gene;Human;Human Development;human disease;Human Genetics;human TNF protein;Image;in vivo;Individual;insight;interdisciplinary approach;Ion Channel;Joubert syndrome;Kidney Diseases;Laboratories;Length;male;Mammalian Cell;Mechanics;Medical;Modeling;Molecular;Molecular Genetics;molecular imaging;Molecular Motors;Morphogenesis;Motor;Mus;Mutate;Mutation;Nematoda;Neurons;novel;novel strategies;Organ;Organelles;Partner in relationship;Phosphatidylinositols;Phosphoric Monoester Hydrolases;PKD2 protein;Play;polycystic kidney disease 1 protein;Post-Translational Protein Processing;Process;Promotor (Genetics);protein function;Proteins;Purkinje Cells;Reagent;receptor;Renal function;Reproducibility;Research;RNA Interference;Role;sensor;Sensory;Shapes;Signal Pathway;Signal Transduction;Stimulus;System;Techniques;Testing;tool;trafficking;Transgenic Animals;Tubulin;Tumor Necrosis Factor Receptor;Tumor Necrosis Factor-alpha;ubiquitin-protein ligase;Ubiquitination;Work;

Abstract: DESCRIPTION (provided by applicant): Cilia are cellular organelles that are important for human development, organ function, and sensation. Defects in cilia formation or function results in devastating human diseases or ciliopathies, including autosomal dominant polycystic kidney disease (ADPKD). ADPKD affects 1/400-1000 individuals, often resulting in end- stage renal disease. In humans, mutations in the polycystin-1 or polycystin-2 receptor-channel complex cause ADPKD. Despite the profound medical importance of cilia in human health, how cilia are specialized in form and function remains poorly understood. This proposed research is focused on uncovering fundamental principals of cilia biology not possible in human studies or easily studied in vertebrate model systems. C. elegans is a transparent, multicellular animal with specialized sensory cilia, features that enable in vivo imaging that is unprecedented in its simplicity and reproducibility. We have developed the tools and reagents to study polycystin localization and function in C. elegans. We will employ genetic, molecular, imaging, biochemical, and electrophysiological approaches to address three specific aims. First, we will dissect polycystin signaling pathways. Next, we will determine the molecular details of mechanisms regulating polycystin localization, focusing on the roles of phosphoinositides and tubulin post-translational modifications. Finally, we will identify new genes regulating PKD-2 localization using transgenic animals that are hypersensitive to neuronal RNAi. This multidisciplinary approach will provide a comprehensive picture of the molecules that influence polycystin channel assembly and trafficking, and will provide critical insight to ciliary receptor trafficking in general. Thus these studies will have important implications in human diseases such as ADPKD in which cilia play an important and central role. PUBLIC HEALTH RELEVANCE: Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common monogenic diseases, affecting 1/400-1000 individuals, and having no treatment or cure. Several human genetic disorders, including ADPKD, share two common features: ciliary localized gene products and kidney cysts. Given that it is prohibitively difficult in humans to study the connection between ciliary protein function, localization, and disease, alternative experimental systems are necessary. In C. elegans it is feasible to study the roles of human disease gene homologs in cilia formation, morphogenesis, and signaling. This proposal is aimed at understanding the molecular mechanisms controlling the subcellular trafficking of the C. elegans ADPKD gene products.

GUSTATORY LOSS AND RECOVERY OF PATIENTS WITH MIDDLE EAR SURGERY

Start Date: 9/1/2011

Award Number: 1R21DC011393-01A1

NIH Funding Organization: NIDCD

Principal Investigator: BRESLIN, PAUL A.S.

Co-PI: ARTZ, GREGORY

FY Total Cost by IC: $229951

Program(s): Anterior;Apoptotic;Axon;axon regeneration;Basic Science;Biopsy;Body Weight;Body Weight decreased;cell type;Cells;Chemicals;chorda tympani;Clinical;Cranial Nerves;Data;Dendritic Cells;Detection;Dissection;Eating;Esthesia;Etiology;Excision;experience;Facial nerve structure;Foliate Papilla;Food;Functional disorder;Fungiform Papilla;Human;human tissue;Immune;Incus structure;Inflammatory;Injury;interest;Ipsilateral;Lateral;Leucocytic infiltrate;Lymphocyte;macrophage;Malleus;Mammals;Measures;Medial;Methods;Microscopic;middle ear;middle ear disorder;Morphology;Natural regeneration;Nature;Nerve;Nerve Crush;nerve supply;Operative Surgical Procedures;Oral cavity;Organ;Otologic Surgical Procedures;Patients;Peripheral;Phenotype;Physicians;Procedures;Process;Psychophysiology;receptor;Receptor Cell;Recovery;regenerative;Reporting;Resected;Risk;Rodent;Severities;Side;Stimulus;Stomach;Stretching;Taste Bud Cell;Taste Buds;Taste Perception;Testing;Time;Tissues;Tongue;tongue apex;Trauma;Tympanic membrane structure;Work;

Abstract: DESCRIPTION (provided by applicant): Taste is the paramount sense governing which foods are accepted or rejected. It is the final arbiter of what goes into the stomach and what is rejected. We believe this is because the taste of food is, in general, the most important marker of its identity and acceptability. To this point, chemosensory clinical patients who present with taste complaints are twice as likely to report weight loss as those who present with olfactory dysfunction (40% vs. 20%) and enjoy food less. We, therefore, wish to understand the loss and recovery of chemosensory taste function in patients who have iatrogenic taste nerve damage during middle ear surgery. The chorda tympani (CT) nerve, which innervates the taste buds of the anterior two-thirds of the tongue, may be damaged during this surgery. As a clinical problem, the physician is unable to predict for their patient the degree or chemospecificity of their potential taste loss or precisely when normal function will return. From a basic research perspective, the proposed work will investigate the potential for a human cranial nerve and receptor end organs to re-grow and regenerate. A better understanding of a fully regenerating organ and tissue has great potential to help us understand how other human tissues, which do not spontaneously regenerate, may be induced to do so. The Aims of this proposal are to explore the loss and recovery of taste perception as a function of the type of CT nerve damage that occurs during surgery and the concomitant changes to human taste buds. PUBLIC HEALTH RELEVANCE: Taste is the final arbiter of what is eaten and what is rejected. Patients who present with taste complaints may lose interest in food and body weight. We wish to understand the loss and recovery of taste function in patients who have taste nerve damage during surgery of the middle ear. We will explore the loss and recovery of taste that occurs during surgery and the concomitant changes to human taste buds.

TRANSIENT BEHAVIORS OF ADAPTING BIOLOGICAL SYSTEMS

Start Date: 8/19/2011

Award Number: 1R01GM100473-01

NIH Funding Organization: NIGMS

Principal Investigator: SONTAG, EDUARDO

Co-PI: Not Applicable

FY Total Cost by IC: $327212

Program(s): Apoptosis;Area;base;Behavior;Biologic Characteristic;biological systems;Biology;Biomedical Engineering;Cell Proliferation;Cellular biology;Chemicals;Chemotactic Factors;Chemotaxis;Colorectal Cancer;Cues;Detection;Development;Embryo;Environment;Epidermal Growth Factor;Fluorescence Resonance Energy Transfer;Fostering;Goals;Homeostasis;Immune system;Individual;insight;Ligands;Link;Mathematics;Measurement;Microfluidics;Molecular;Morphogenesis;Motion;Nonlinear Dynamics;Organism;pathogen;Pathway interactions;Pattern;Play;Primary carcinoma of the liver cells;Property;Proteins;response;Role;Sensory;Signal Pathway;Signal Transduction;spatiotemporal;Stem cells;Stimulus;Stress;System;Systems Biology;Technology;Testing;theories;Time;Tissues;tool;Tumor Angiogenesis;Uncertainty;Variant;Work;Wound Healing;

Abstract: DESCRIPTION (provided by applicant): The behavior, adaptability, and survival of organisms, including pathogens or immune system components, depend critically upon the capability to formulate appropriate responses, at different time scales, to fluctuating temporal patterns of environmental cues such as ligand concentrations or stresses. Traditionally, mathematical studies have focused on constant environments and steady-state behaviors, not on naturally occurring signals and transient behavior. The ultimate goal of this work is to investigate, both theoretically and experimentally, the characteristics of biological responses to time-varying signaling and what information is encoded in them. The primary focus will be on the study of invariance with respect to sensory symmetries, with an initial emphasis on fold-change-detection (FCD), a property that confers signaling systems robustness to scale uncertainty. Recent molecular cell biology studies have experimentally discovered FCD behavior in prokaryotic chemotaxis pathways, allowing responses to broad ranges of chemoattractant concentrations, and in the eukaryotic epidermal growth factor ERK and the Wnt signaling pathways, allowing robustness of behavior in the face of large variations in protein abundances. One aim of this work is to develop new, and to significantly expand existing, theory of FCD and other symmetries. This aim will involve foundational work in areas of mathematics ranging from control theory to group representations. Broader significance will include new insights in nonlinear dynamics and systems biology, as the properties studied are emergent synthetic behaviors, not explainable easily from the properties of individual chemicals in isolation. A complementary aim is to develop an experimental platform to study transient behaviors, particularly in chemotaxis. New experimental tools will be established, based on microfluidic technology and on molecular level analysis of intracellular signaling (FRET), to afford unprecedented spatiotemporal control of stimuli during measurements and to allow the effective testing of mathematical predictions. Broader significance will be in fostering the development of new microfluidic technology of wide applicability in biology and biomedical engineering. PUBLIC HEALTH RELEVANCE: Chemotaxis plays a key role in bacterial motion, as well as in embryonic morphogenesis, tissue organization, tumor angiogenesis, and wound healing. Activity of the ERK pathway controls cell proliferation, differentiation, and apoptosis. The Wnt signaling pathway plays a role in embryonic patterning and stem cell homeostasis, and its disregulation has been linked to colorectal cancer and hepatocellular carcinoma.

EPIGENETIC TRANSGENERATIONAL EFFECTS OF ENDOCRINE DISRUPTORS VIA FEMALE GERM LINE

Start Date: 8/12/2011

Award Number: 1R21ES017847-01A1

NIH Funding Organization: NIEHS

Principal Investigator: UZUMCU, MEHMET

Co-PI: Not Applicable

FY Total Cost by IC: $231125

Program(s): Aberrant DNA Methylation; Address; Adult; Affect; Aging; Anxiety; Apoptosis; base;Behavior;beryllium trifluoride;bisulfite;Breeding;Chemical Exposure;Child;demethylation;density;Development;Disease susceptibility;DNA Methylation;DNA Methyltransferase;DNA Modification Methylases;DNA Sequence;Dose;Embryo;Endocrine Disruptors;Endogenous Retroviruses; Environment;environmental stressor;Epigenetic Process;Event;Exhibits;Exposure to;Female;Fertility;fetal;folliculogenesis;follow-up;Future;Future Generations;Gene Expression;Generations;Genes;Germ Cells;Germ Lines;Global Change;Goals;Gonadal structure;grandchild;H19 gene;Health;Health Care Costs;imprint;in utero;Inbreeding;Incidence;Individual;innovation;Knowledge;Lead;Life;Link;male;Mediating;Mediator of activation protein;Methods;Methoxychlor;Methylation;Modeling;Morphology;Neonatal;novel;Onset of illness;Oocytes;Ovarian;Ovary;Ovulation;Parents;Partner in relationship;Pathology;Pattern;Perinatal Exposure;Periodicity;Pesticides;Phenotype;Play;postnatal;prevent;programs;Puberty;public health medicine (field);Rattus;reproductive;reproductive function;Research;Retrotransposon;Reverse Transcriptase Polymerase Chain Reaction;Risk;Role;sex;Sex Ratio;sperm cell;Testing;Time;TimeLine;Tissues;Toxic Environmental Substances;vinclozolin;Woman;Women's Role;

Abstract: DESCRIPTION (provided by applicant): It is well established that exposure to environmental stressors or toxicants such as endocrine-disrupting chemicals (EDCs) during in utero and/or neonatal development can result in adult-onset diseases. Such detrimental changes have been linked to aberrant DNA methylation patterns as well as other epigenetic mechanisms. It is of great concern that recent studies have suggested that such disease susceptibilities can be transmitted via epigenetic mechanisms to subsequent generations that were never directly exposed to the EDCs. For example, testicular abnormalities were transmitted to F2-F4 generation males via the male germ line after in utero exposure to an EDC. However, these effects were found to be sex-specific since F2-F3 females exhibited milder phenotypes that required exposure of both parents to the EDC. In this proposal, our objective is to address the critical issue of whether the female germ line is involved in such transgenerational (TG) effects of EDC exposure. Methoxychlor (MXC) is an EDC that has actions mimicking those of many other EDCs in the environment and was shown to cause epigenetic changes in the gonad; it will therefore be used as a model EDC in this proposal. Our hypothesis is that exposure to MXC during the critical window of development, defined here as the period of gonad differentiation through oocyte epigenetic reprogramming, leads to TG effects in the ovary that are mediated by epigenetic changes in the oocyte. To test our hypothesis, the proposal has two interrelated but distinct specific aims: (1) Determine somatic TG epigenetic effects of MXC in the ovary of F3 generation following exposure of F1. F1-generation inbred female rats will be treated with 0, 20 5g/kg/day MXC, or 50-100 mg/kg/day MXC between E11 and PND7. These females will be bred with untreated males, and F2-F4 females will be generated. Initially, ovarian morphology and gene expression and fertility will be assessed only in the F3 females since observation of TG effects in F3 generation is the minimum requirement for establishing TG effects. Gene expression changes and global and gene-specific DNA methylation changes will be investigated using conventional methods as well as array-based approaches. (2) Investigate the epigenetic changes induced by MXC in F1/F2 oocytes that may lead to TG effects in F3. DNA methylation patterns in imprinted genes and parasitic DNA sequences such as retrotransposons in oocytes will be analyzed, as aberrant methylation patterns in these sequences can be pathogenic. Oocytic gene expression patterns will also be examined; any correlation between the changes in gene expression and DNA methylation will be elucidated. The proposed study will be the first to investigate the epigenetic TG effects of EDCs transmitted via the female germ line beyond the F2 generation. We recognize the considerable risk associated with this study, but implications to the health of present and future generations are significant given the widespread presence of EDCs in the environment. A mechanistic understanding of the TG epigenetic actions of MXC will broaden our knowledge of how EDCs, in general, disrupt normal epigenetic programming in females, and facilitate the development of strategies to prevent and treat EDC-induced TG abnormalities. PUBLIC HEALTH RELEVANCE: Exposure to environmental endocrine-disrupting chemicals during development can harm the exposed individuals, and this effect can be passed on to their children and grandchildren through epigenetic alterations within the germ cells. The objectives of this research are to investigate the transgenerational epigenetic effects of endocrine disruptors in females and to understand the mechanisms by which these compounds affect the germ cells and transmit these effects to subsequent generations. A better understanding of transgenerational epigenetic effects of endocrine disruptors has direct implications for public health because such an understanding can help us prevent and ameliorate these effects, thereby reducing future health costs.

SUMMER RESEARCH TRAINING IN ENVIRONMENTAL HEALTH SCIENCES

Start Date: 8/9/2011

Award Number: 1R25ES020721-01

NIH Funding Organization: NIEHS

Principal Investigator: LASKIN, DEBRA LYNN

Co-PI: Not Applicable

FY Total Cost by IC: $57510

Program(s): Achievement;Advertising;Applications Grants;Area;Awareness;base;Basic Science;Biochemical;Biomedical Research;carcinogenesis;career;career development;Case Study;Chemoprevention;Commit;Community Health Education;Community Outreach;Core Facility;Dentistry;design;Development;Disadvantaged;Disease;Ecology;Educational aspects;Educational workshop;Endocrine disruption;Engineering;Ensure;Environmental and Occupational Exposure;Environmental Exposure;Environmental Health;Environmental Protection;Evaluation;experience;Exposure to;Faculty;Fellowship;Fellowship Program;Funding;Goals;Government;Graduate Education;Grant;Health Educators;Health Sciences;Immunotoxicology;improved;Individual;Industry;Institutes;Instruction;Joints;Laboratories;Letters;literacy;medical schools;Medicine;meetings;member;Mentors;Mentorship;Methodology;National Institute of Environmental Health Sciences;neurotoxicity;New Jersey;news;novel;Occupational Health;Pharmacy Schools;Positioning Attribute;Preparation;Professional Education;programs;public health relevance;Recommendation;Research;research and development;Research Infrastructure;Research Personnel;Research Project Grants;Research Training;Resources;responsible research conduct;Science;Series;Signal Transduction;skills;Structure;Students;Technology;Time;Toxicology;Training;Training Support;Translational Research;United States National Institutes of Health;Universities;Wood material;

Abstract: DESCRIPTION (provided by applicant): The summer program proposed in this grant application will introduce undergraduate students to the field of environmental health sciences as part of the existing Summer Undergraduate Research Fellowship (SURF) at Rutgers University. The purpose of SURF is to provide undergraduates with 10-week basic science and translational research experiences in preparation for careers in biomedical research. Since its inception, SURF has provided 30 undergraduates with summer research experiences and this proposal will establish an additional 10 fellowships for students to conduct research in toxicology and environmental sciences. Strong applications from talented students will be identified by scholastic achievement, letters of recommendation, and compelling personal statements. Recruitment strategies have been designed to attract students from disadvantaged and under-represented backgrounds into the fellowship program. Under the guidance and instruction of NIH-funded investigators, SURF students will conduct hands-on research projects in laboratories focused on environmental health sciences. Students will meet with mentors weekly to receive one-on-one training, ensure continued development of research skills, and evaluate progress of scientific projects. This program draws upon the resources, infrastructure, and expertise of faculty at Rutgers University and UMDNJ- Robert Wood Johnson Medical School who are members of the Environmental and Occupational Health Sciences Institute, and part of our joint NIEHS T32 training grant and NIEHS Center for Environmental Exposures and Disease. Students will also participate in weekly seminars and workshops on the topics of career development, responsible conduct of research, and environmental research. In addition, students will discuss topics 'in the news' that are related to environmental exposures, which will promote overall science literacy. They will also tour core facilities established as part of the NIEHS Center and be instructed in state-of- the-art technology for biomedical research. Students will make a 20-minute PowerPoint presentation of their research at the end of the summer. The SURF Directors and Project Managers have developed an evaluation plan to ensure the continued development and improvement of the fellowship program over subsequent years. Public Health Relevance: The structured hands-on research, didactic, and career activities in this proposal are designed to encourage and prepare undergraduate students for graduate and professional education in environmental health sciences.

NATURAL PRODUCTS IN THE PREVENTION OF CANCER

Start Date: 8/1/2011

Award Number: 1R13CA159733-01A1

NIH Funding Organization: NCI

Principal Investigator: SUH, NANJOO

Co-PI: PEZZUTO, JOHN M

FY Total Cost by IC: $5000

Program(s): Achievement;Agreement;Anniversary;anticancer research;Area;Biological;Biological Assay;Biological Factors;cancer chemoprevention;cancer prevention;cancer therapy;carcinogenesis;Chemistry;Chemoprevention;Chemopreventive Agent;Clinical;Collaborations;Collection;Communication;Complex;Data;design;Development;Discipline;Disease;disorder prevention;drug candidate;Environment;Epidemiology;Fostering;Future;General Population;Goals;graduate student;Hawaii;Health;Human;improved;Incidence;interest;International;Island;Lead;lectures;Malignant Neoplasms;Medicinal Plants;meetings;Modification;Mortality Vital Statistics;North America;novel;Parents;Participant;Pharmaceutical Preparations;Phytochemical;Plants;Play;pre-clinical;prevent;Prevention;Prevention approach;Prevention Research;Prevention strategy;Preventive;public health medicine (field);Publications;Publishing;Research;Research Personnel;Resources;Role;Scientist;Secure;Series;Societies;Staging;success;symposium;Thinking, function;United States National Institutes of Health;Vaccines;Work;

Abstract: DESCRIPTION (provided by applicant): Natural products provide diverse resources in the development of new drugs for cancer treatment and prevention. Cancer chemoprevention is defined as an approach for the prevention, delay, or reversal of carcinogenesis. To discover and develop new cancer chemo preventive agents, many natural product researchers and medicinal chemists have focused on the exploration of plants with biological activity in bioassays designed to detect potential cancer preventive agents and the isolation of natural products as lead molecules, as well as modification of the active components for further synthesis. Recent advances in the chemistry of novel bioactive natural products provide improvement and development of new drugs. For the purpose of prevention of complex diseases such as cancer, there have been tremendous efforts of diverse groups of scientists working together to develop new and safe drugs from naturally-occurring resources. In order to review recent progress in this area and help to define a path to the future, we have assembled a group of internationally renowned experts in cancer prevention to participate in a symposium and present their recent work. Principal topics of the session will be potential chemopreventive agents developed from natural products and medicinal plants. The meeting will cover preclinical, clinical and epidemiological data in the area of cancer prevention research with compounds, either naturally occurring or derived from natural products components. There will be ten invited speakers for the session. Further discussion on the future direction of natural product research in the area of cancer prevention will be of great interest. At the start of the symposium, a lecture will be presented to overview the lectures to come, highlight the challenges of the field, and set the stage for the proceedings to come. The objective of the session is to critically evaluate progress and begin to chart of course for future work. Each lecture will include questions and answers and discussion. At the end of the session, a round table discussion will be moderated that will include each of the speakers and the audience. The symposium will be conducted as part of the 50th Annual Meeting of the Phytochemical Society of North America that will be held during the period of December 10-14, 2011, on the Kohala Coast of the Big Island of Hawaii. We will employ all of the resources assembled by the meeting organizers to assure the success of this symposium. The audience will be diverse and international. The venue is centrally located to facilitate East/West interaction. Very favorable hotel rates have been secured to promote broad attendance including graduate students and other trainees. It is expected the attendees will reflect broad diversity. The proceedings of the symposium will be published by Springer in their renowned series entitled Topics in Current Chemistry (impact factor 5.270; 2009). Each chapter as well as the preface will acknowledge presentation at the meeting and support provided by the NIH. This will be a landmark achievement in the field of cancer chemoprevention. PUBLIC HEALTH RELEVANCE: Prevention of cancer is a safe and effective way to reduce human suffering. In principle, cancer chemoprevention could play a role as dominant as vaccines in the prevention of disease. Nonetheless, despite the great promise of this approach, progress has been hampered. We propose to organize a meeting that will bring outstanding researchers from both cancer prevention and natural products research field together. The objective is to increase communication, disseminate new findings, prepare a platform for increased collaborations in the use and development of novel components and their derivatives from natural products as potential cancer preventive agents, and to explore and define barriers and ways to overcome these barriers. A prevention approach with natural products is an important part of cancer prevention strategies for improving general public health. We believe that the conference may contribute to achieving the goal of the NIH, and will provide a better understanding and approaches to reduce cancer incidence and mortality in the coming decades.

R25 IMSD MINORITY BIOMEDICAL RESEARCH SUPPORT PROGRAM (MBRS)

Start Date: 7/1/2011

Award Number: 1R25GM096161-01

NIH Funding Organization: NIGMS

Principal Investigator: KOMISARUK, BARRY RICHARD

Co-PI: Not Applicable

FY Total Cost by IC: $624224

Program(s): Achievement;Behavioral;Behavioral Research;Biomedical Research;career;Communities;Consult;Counseling;Data;design;Development;Doctor of Medicine;Doctor of Philosophy;Drug Formulations;Educational workshop;Ensure;Evaluation;experience;Feedback;Fostering;Goals;improved;Institution;Interview;Knowledge;Lead;Mainstreaming (Education);meetings;Mentors;metropolitan;Minority;Monitor;Motivation;New Jersey;New York City;Outcome;Participant;Performance;Personal Satisfaction;Phase;Play;Population;Positioning Attribute;Process;Process Assessment (Health Care);programs;psychologic;Publications;Questionnaires;Recommendation;Recruitment Activity;Research;Research Support;Research Training;Role;Schools;Self Efficacy;Services;skills;Students;symposium;Testing;Time;Time Management;Training;Underrepresented Minority;Universities;

Abstract: DESCRIPTION (provided by applicant): The goal of this new proposed IMSD Program is to increase the number of minority students who enter careers in biomedical and behavioral research in which they are under-represented. Our proposed comprehensive program will play a significant role in overcoming the still-existing disparities, between minority and non-minority undergraduate and doctoral students at Rutgers, in their progressing into careers in biomedical/behavioral research. Through accomplishing this goal, our proposed IMSD Program will improve our overall Institutional outcomes. To accomplish this goal, our objectives are to provide the undergraduate and doctoral students in our Program with the academic and research skills, self-efficacy, motivation, and credentials that are essential to developing their competitiveness toward entering the next phases of their career - i.e., doctoral programs or postdoctorates, respectively. Thus, our proposed IMSD Program will: ' Provide the undergraduate and doctoral students with a carefully-mentored, rigorous research experience that includes supervised conceptualization, formulation, performance, analysis, presentation at professional scientific conferences, and publication of their own research or doctoral dissertation project; ' Provide a sequentially-coordinated, closely and continuously monitored spectrum of academic and personal support components to promote the students' highest academic performance levels, including, e.g., academic enrichment classes in gate-keeper courses, test-taking skills workshops, psychological counseling services, career guidance workshops, mentored graduate school and postdoctoral application processes, all designed to develop the students' professional self-efficacy, ensure their personal well- being and retention through to completion of their undergraduate or doctoral training on campus, and facilitate their entry into doctoral or postdoctoral programs; ' 'Mainstream' the students into the academic campus community by articulating our IMSD Program with other relevant University programs, prominent among which is the new 'Garden State LS-AMP Program.' The lead institution for this major new Program is Rutgers-Newark, so our campus will play a major role in its development, and it would be a major new interaction opportunity for our IMSD Program. ' Cast a wide net to recruit potential IMSD students -- especially doctoral students, who are particularly underrepresented in the biomedical and behavioral-related departments on our campus -- from multiple venues both on and off-campus, including the new Garden State LS-AMP Program and its participating New Jersey institutions, and the greater metropolitan New York City institutions with significant under- represented minority student populations totaling several hundred IMSD-eligible students in relevant major fields. ' Monitor continuously the performance of our IMSD Program via data gathering, analysis, and review of our quantitative goals by the IMSD PD and staff, and regular meetings with our IMSD participants, toward eliminating any gaps in performance between our IMSD students and the students in the university community at large. Assisting in this assessment process will be annual formal evaluations by SageFox Consulting, which will perform interviews, provide and analyze questionnaires, and provide feedback and recommendations to the students, mentors, advisory board, and PD of our Program. ' Among the quantitative expected outcomes for our IMSD undergraduate students: at least 90% of the undergraduates accepted into our IMSD Program will earn the baccalaureate, of whom at least 70% will proceed to enter a Ph.D. or Ph.D./M.D. program. And at least 90% of the doctoral students accepted into our IMSD Program will earn the Ph.D., of whom at least 80% will proceed to a postdoctoral position. PUBLIC HEALTH RELEVANCE: The goal of this new proposed IMSD Program is to increase the number of minority students who enter careers in biomedical and behavioral research in which they are under-represented. Our proposed comprehensive Program will play a significant role in overcoming the still-existing disparities, between Minority and non-Minority undergraduate and doctoral students in our Institution, in their progressing into careers in biomedical/behavioral research. Through accomplishing this goal, our proposed IMSD Program will improve our overall Institutional outcomes. To accomplish this goal, our objectives are to provide the undergraduate and doctoral students in our Program with the academic and research skills, self-efficacy, motivation, and credentials that are essential to developing their competitiveness toward entering the next phases of their career - i.e., doctoral programs or postdoctorates, respectively.

ENDOSOME TO GOLGI RETROGRADE RECYCLING IN C.ELEGANS

Start Date: 7/1/2011

Award Number: 1F32GM096599-01

NIH Funding Organization: NIGMS

Principal Investigator: NORRIS, ANNE

Co-PI: Not Applicable

FY Total Cost by IC: $51326

Program(s): Alleles;Amino Acids;Apoptotic;Architecture;ATP phosphohydrolase;base;Binding (Molecular Function);Biological Assay;Caenorhabditis elegans;Cell Adhesion Molecules;Cell physiology;Cell Polarity;Cell Surface Proteins;Cells;Clathrin;Complex;Data;Defect;Degradation Pathway;Development;Disease;DNA;E-Cadherin;Early Endosome;Endocytosis;Endosomes;Energy Transfer;Epithelial Cells;Equilibrium;Event;Generations;Genes;Genetic;genetic analysis;Genetic Epistasis;Genetic Screening;Goals;Golgi Apparatus;Homeostasis;Immunoprecipitation;in vivo;insight;Intercellular Junctions;Intestines;Knowledge;Laboratories;Late Onset Alzheimer Disease;Left;Link;Lipids;Location;Lysosomes;Maintenance;Malignant Neoplasms;Mammalian Cell;Mammals;Mediating;Membrane;Membrane Protein Traffic;Modeling;Molecular;Molecular Chaperones;Multivesicular Body;mutant;novel;Nutrient;Organism;Pathway interactions;Process;Protein Binding;Protein Region;protein transport;Proteins;Recycling;Regulation;Research Design;research study;retrograde transport;RNA Interference;Role;Route;Saccharomyces cerevisiae;Screening procedure;Shiga Toxin;Signal Transduction;Signaling Molecule;Sorting - Cell Movement;Technology;Tertiary Protein Structure;Testing;Tissues;trafficking;Transmembrane Transport;uptake;Work;Yeasts;

Abstract: DESCRIPTION (provided by applicant): Title of Project: Endosome to Golgi Retrograde Recycling in C. elegans Background: Cells and tissues establish and maintain their unique architectures in large part through the tight regulation of protein and membrane transport. Key aspects of this process include the decision to recycle or degrade a given molecule, particularly signaling and adhesion molecules, a process that goes awry in many cancers. After endocytosis lipids and trans membrane cargo enter the early endosome for sorting. Cargo may be directed to degrade via targeting to the lysosome, recycled to the PM directly or through the recycling endosome, or sorted for delivery to the Golgi via the retrograde route. A number of observations hint that these routes compete with each other for various cargo, and that the decision to recycle or degrade is a key point of regulation. Our focus is on the much underappreciated branch point between the retrograde route and the degradative route to the lysosome. It has been suggested that competition between these two routes is mediated by microdomains that form on the early endosome and multivesicular body. The clathrin/HGRS-1(Hrs)/ESCRT complexes form one microdomain that promotes degradation via the lysosome, and the retromer/RME-8 complex forms a competing microdomain and promotes the retrograde pathway, recycling cargo from the endosome to the Golgi. The goal of our proposal is to better understand the molecular mechanisms controlling the decision of cargo to traffic via the retrograde or degradative pathways, as well as to find new retrograde components. Objective/Hypothesis: Our lab has recently made advances in our understanding of the molecular mechanism of this competition between the clathrin/HGRS-1(Hrs)/ESCRT and RME- 8/retromer microdomains. Our data suggest that SNX-1, a retromer component, potentiates the binding of RME-8 to the chaperone HSP-1(Hsc70), an interaction that antagonizes the buildup of endosomal clathrin and the associated degradative subdomain via clathrin uncoating activity. Furthermore preliminary evidence suggests that HGRS-1(Hrs) directly competes with this clathrin uncoating process potentially by titrating SNX-1 away from RME-8, leaving clathrin free to build up and promote lysosomal degradation. We aim to test this microdomain maintenance hypothesis, as well as dramatically extend our knowledge of the retrograde pathway in a multicellular organism by large-scale RNAi-based screening. Specific Aims: (1) To determine the functional consequence of SNX-1/RME-8 interaction (2) Distinguish between two models to explain the HGRS-1(Hrs)/SNX-1 interaction, especially as it relates to RME-8/HSP-1(Hsc70) mediated endosomal sorting and (3) To identify novel components of the retrograde sorting pathway. Study design: We propose to analyze our competition model using a combination of in vivo rescue experiments with interaction impaired mutants of RME-8, epistasis analysis, and Fvrster resonance energy transfer (FRET) analysis. Additionally we propose a simple yet powerful large scale RNAi screen to identify novel retrograde pathway components. Significance: All of the components of retrograde and lysosomal sorting we are studying are conserved in mammals. Defects in retrograde transport are linked to late-onset Alzheimer's disease, E-cadherin mis-localization, defects in the generation of Wnt signaling gradients, and defects in the clearance of dead apoptotic cells. Additionally the retrograde sorting route is the mode of entry for Shiga Toxin. We believe that a better understanding the molecular mechanism that target MIG-14 to its proper location within the cell, as well as identifying new genes that have a role in this process, will help us to understand the underlying molecular events leading a vast array of disease states. PUBLIC HEALTH RELEVANCE: Endocytosis and endosomal sorting involves the uptake of lipids and proteins from the cell surface followed by their proper distribution in the cell. This 'trafficking' is critical for diverse cellular functions such as signaling, cell-cell junction maintenance, nutrient uptake, and development. Defects in transport are linked to late- onset Alzheimer's disease, defects in cellular polarity (knowing its top from its bottom) and defects in the clearance of dead cells.

MARIJUANA CUES, AROUSAL AND THE CENTRAL AUTONOMIC NETWORK

Start Date: 7/1/2011

Award Number: 1R03DA031060-01

NIH Funding Organization: NIDA

Principal Investigator: BATES, MARSHA E

Co-PI: Not Applicable

FY Total Cost by IC: $227652

Program(s): Abstinence;Activation Analysis;addiction;Addictive Behavior;Affect;Alcohol or Other Drugs use;Alcohols;Amygdaloid structure;Area;Arousal;Autonomic nervous system;Back;Baroreflex;Behavior;Biofeedback;Biological Neural Networks;Brain;Brain imaging;Brain region;Brain Stem;Cardiovascular Physiology;Cardiovascular system;Chronic;Cognitive;cue reactivity;Cues;Data;design;drug craving;drug of abuse;Drug usage;Drug Use Disorder;Electrocardiogram;emerging adult;Emotional;emotional experience;emotional reaction;Emotions;Environment;experience;feeding;Fingers;Foundations;Functional Magnetic Resonance Imaging;Goals;Graph;Human;improved;Individual;interest;Intervention;Knowledge;Learning;Link;Marihuana;Marijuana Smoking;marijuana user;Medial;memory process;Methodology;Modeling;Nervous System Physiology;Neuraxis;neuroimaging;neuromechanism;neurophysiology;novel strategies;operation;Participant;Peripheral Nervous System;Persons;Pharmaceutical Preparations;Physiologic pulse;Prefrontal Cortex;Prevention;Process;Psychophysiology;public health medicine (field);Regulation;Relapse;relating to nervous system;Relative (related person);Research;Research Personnel;research study;Respiration;response;Role;skills;Smoker;Smoking;Stimulus;Structure;Support System;System;Time;Training;Translational Research;Visceral;Vision;Visual;Work;

Abstract: DESCRIPTION (provided by applicant): This application is in response to PAR-09-073 to facilitate the entry of two addiction researchers into the area of functional magnetic resonance imaging (fMRI). We are currently characterizing moment-to-moment changes in cardiovascular arousal in response to emotionally-valenced and appetitive visual cues. This research focus is of substantial public health significance due to the fundamental role of cue reactivity and drug craving in motivating alcohol and drug seeking, use, and relapse following treatment. Visceral changes in autonomic nervous system (ANS) functions contribute significantly to the phenomenal experience of emotional arousal through extensive ANS and central nervous system (CNS) interconnections and the baroreflex. Yet, understanding of dynamic mechanisms that control cue reactivity has been hampered by conceptualizing ANS functions in isolation from the neural system(s) within which they operate. We seek to apply a neurophysiological systems approach to characterize how intra-individual changes in functional, regulatory aspects of emotional arousal in response to environmental cues are integrated across the ANS and CNS. The specific aims are to gain expertise in fMRI methodology and analysis, and to apply this expertise to study a dynamic neural system that modulates cardiovascular response to drug cues. As proof-of-concept, an experiment merging psychophysiological and neuroimaging methodologies is proposed as a fMRI and connectivity analysis learning vehicle (Aim 1) and to examine substantive differences in integrated brain and cardiovascular system response to marijuana-related picture cues in regular marijuana users compared to matched controls who do not use marijuana (Aim 2). fMRI data during cue exposure will be gathered using a 3-Tesla magnet, and the spread of activation between brain regions involved in arousal modulation will be examined using brain connectivity analysis. Activation and connectivity of three brain regions (medial prefrontal cortex, amygdala, brain stem) will be quantitatively linked to cardiovascular modulation within a graph theoretical connectivity model. If successful, this application will yield (1) preliminary support for a new approach wherein ANS reactivity can be linked dynamically (in a time-varying manner, not as a correlation) to neural systems that modulate cue reactivity, (2) new knowledge about how activation in three brain regions interrelates with baroreflex modulation of cardiovascular response to cues, and (3) the groundwork for translational research aimed at evaluating whether biofeedback interventions that improve specific cardiovascular functions can be used to affect inhibition in brain systems that control arousal in response to drug cues. PUBLIC HEALTH RELEVANCE: Cues in the environment, such as the sight of someone smoking, can produce a powerful emotional reaction that increases the likelihood that susceptible individuals will seek or use a drug of choice, even following periods of abstinence. The visceral experience of emotional arousal is an important component of reactivity to drug cues. This application seeks to characterize how the autonomic nervous system works together with the brain to control visceral response to marijuana cues.