Anantha Shekhar - US grants

Anxiety disorders are the most prevalent psychiatric disorders and appear to be a heterogenous group of disorders. The neurobiology of anxiety disorders is still poorly understood. However, several limbic structures have been consistently implicated in the neurobiological mechanisms underlying anxiety such as the hypothalamus, amygdala, and others. Several brainstem structures such as the locus ceruleus and the raphae nuclei have also been implicated in the etiology of anxiety disorders. The posterior hypothalamus appears to be a key region in the expression of anxiety with a neuronal circuit capable of eliciting the physiological and behavioral components of anxiety, especially resembling a panic attack. Many CNS regions implicated in anxiety, such as the amygdala and locus ceruleus appear to interact with the hypothalamus and elucidation of these interactions will significantly expand our understanding of anxiety disorders. This study will examine these mechanisms by local microinjection of drugs and pharmacologically manipulating neurotransmitter systems in rats and studying the effects of these procedures on heart rate, respiration, blood pressure, catecholamine levels and locomotion as well as three laboratory tests of anxiety that appear to regulate anxiety at each of the above mentioned CNS sites. Specifically, this research proposes to: 1) characterize the effects of modifying GABA neurotransmission in the posterior hypothalamus of rats on anxiety with particular emphasis on the possible involvement of this region in the etiology of panic disorders; 2) study the role of amygdala in anxiety local pharmacological manipulations of locus ceruleus on anxiety and determine the interaction of posterior hypothalamus with this region; 3) study the effects of pharmacological manipulations of locus ceruleus on anxiety and determine the interaction of the posterior hypothalamus with the locus ceruleus. The results of the proposed study will increase our understanding of the anxiety disorders and might help us devise better pharmacological strategies for their treatment.

DESCRIPTION (provided by applicant): Panic disorder is a severe anxiety disorders characterized by significant disability. Rats in which GABA inhibition is chronically disrupted in the dorsomedial hypothalamus/perifornical (DMH/PeF) region, exhibit heightened anxiety and panic-like responses, defined as increases in heart rate (HR), mean arterial pressure (BP), respiratory rate (RR), and anxiety as measured by the social interaction (SI) test, following exposure to subthreshold cues such as 0.5 M sodium lactate and 7.5% CO2, agents known to provoke panic attacks in subjects suffering from panic disorder. During the last funding period, we have extensively characterized the afferent pathways for the lactate stimulus, demonstrated some of the regulatory mechanisms within the DMH, and identified the efferent targets of the DMH that are implicated in the panic-like response. The goal of this competitive renewal (MH 52619) is to further elucidate the pathways and neurotransmitters involved in the different components of panic response using pharmacological, functional neuroanatomical, and molecular studies. Overall Hypothesis of the work is that disruption of GABA inhibition in the DMH/PeF region of rats induces a 'panic-prone'state, as a result of pathological activation of a select group of glutamate/peptidergic projection neurons, most prominently orexin/dynorphin A (ORX/DYN) positive cells. This results in aberrant stimulation of a number of efferent targets from the DMH. During this funding period, we will study the pathways involved in activating the bed nucleus of the stria terminalis (BNST) to result in anxiety-like responses and specific brain stem projection targets such as the nucleus tractus solitarius (NTS) in causing inhibition of parasympathetic and activation of sympathetic pathways to increase HR and BP following lactate infusions in these panic-prone rats. We will test these hypotheses with experiments using neuronal immunohistochemical studies;systemic injections of ORX and other peptide receptor antagonists;targeted lesioning of the ORX neurons in the DMH/PeF;measuring the changes in pre-proORX (ppORX), proDynorphin (pDYN) and other peptide mRNA expressions using RTPCR;and acute ppORX and/or pDYN gene knockdown with siRNA, within the DMH. We will study the efferent sites with infusions of pharmacological agents, gene silencing using siRNA as well as neuroanatomical techniques. Finally, we will study the role of local versus extrinsic GABA neurons by GAD-67/65 gene silencing in the DMH/PeF region.

[unreadable] DESCRIPTION (provided by applicant): In this application, we propose to investigate the specific neural systems underlying the mechanisms that may lead to long-term sensitization of one of the 'anxiety regulating networks'. The overall hypothesis for the current proposal is that the state of 'anxiety' (measured by behavioral an physiological responses) in an animal is dependent upon sensitization of a distributed forebrain neural network (that includes the basolateral and central nuclei of the amygdala, bed nucleus of the stria terminalis and others) and a topographically organized serotonergic system in the caudal dorsal raph_ nucleus. The experiments described below have been designed to test this hypothesis under four Specific Aims: 1) To demonstrate that the critical components of the proposed neuronal network, specifically, the corticotropin releasing factor (CRF) containing neurons of BLA, CeA, BNST, and the serotonergic neurons of the dorsal raph_nucleus (DRN), are activated (as measured by c-fos activation) in a variety of 'anxiety' states. 2) To determine that repeated stimulation of CRF receptors, either CRF type 1 or 2 receptors (CRFR1 and CRFR2, respectively) within the above network results in 'priming' of chronic anxiety/stress states. 3) To characterize, using combined anatomical and electrophysiological techniques, the effects of stimulating the CRF receptors on the 5-HT neurons of the caudal DRN in control versus chronically anxious (such as those after repeated CRF stimulation) rate. 4) To determine the effects of early life events such as neonatal lipopolysaccharide (LPS) stress on the BLA, CeA, BNST and DRN network functions. All these studies will be utilizing the following methods: a) whole animal behavioral and physiological responses; b) microinjection and immunohistochemical studies to elucidate pathways; c) patch-clamp studies to determine electrophysiological changes at the neuronal level; as well as d) pharmacological and molecular methods to understand regulation of messenger RNA levels that may contribute to long-term changes in cellular function. This proposal is a direct result of a NIMH research network development grant, HPA Regulation: Cross-Disciplinary Research Networks (RO3 MH 60825) and has provided us with an opportunity to demonstrate the feasibility of this proposed international collaboration. [unreadable] [unreadable]

This subproject represents an estimate of the percentage of the CTSA funding that is being utilized for a broad area of research (AIDS research, pediatric research, or clinical trials). The Total Cost listed is only an estimate of the amount of CTSA infrastructure going towards this area of research, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Overarching Goal of the Indiana Clinical and Translational Science Institute (CTSI) is to transform the Darticipating institutions to create an environment that facilitates the conduct of biomedical research. Mission Statement: "To increase translational biomedical research and improve the health of the people of Indiana and beyond." To this end, we have developed new mechanisms to accelerate translational research, including enhanced educational programs to train translational researchers, a newly designed community engagement activity to produce effective and bidirectional community partnerships, streamlining of all available research infra- structure to accelerate translational projects, and partnering with commercial and philanthropic organizations in Indiana. Throughout all of these efforts runs the critical link of our medical informatics program, enabling all parties to interact in a facile and responsive and prompt manner. Thus, the Indiana CTSI brings together the resources of an entire state. In short, it will provide to the national network of CTSAs: A true statewide laboratory to experiment with innovative methods aimed at transforming research in biomedical sciences, health economy, health care delivery, and health policy. The 5 Specific goals of the Indiana CTSI: I. Create Translational Research Acceleration Programs and Support Pilot Projects by providing investigators and consumers with strategic leadership and mentorship to identify, evaluate, and support innovative and important pilot research at each step of the translational cycle. . Train a new cadre of Translational Researchers by strengthening existing programs and creating new ones to educate trainees and engage faculty in the translational sciences. HI. Foster Robust Community Engagement by creating novel programs with bidirectional participation (i.e., from academia to the community and back again), such as the Indiana Community Health Enhancement Program (CHEP) and pilot programs in Health Econmics, and Rural and Global Health. IV. Build Facile and Comprehensive Research Resources and Technologies by transforming the existing and new research infrastructure into innovative programs such as the Participant and Clinical Interaction Resources (PCIR), Translational Technology Resources (TTR). Research Ethics, Biostatistics and Design Program (BDP), and uniquely, biomedical engineering and bio-nanotechnology to facilitate health research. V. Leverage the Resources of the Greater Indiana Community by connecting to a broad array of resources from multiple partner institutions throughout the state to realize the full potential of the Indiana CTSI.

Clinical Trials; Clinical Trials, Unspecified; Infrastructure; Research Infrastructure; clinical investigation

Infrastructure; Pediatric Research; Research Infrastructure

The Overarching Goal of the Indiana Clinical and Translational Science Institute (CTSI) is to transformthe Darticipating institutions to create an environment that facilitates the conduct of biomedical research. Mission Statement: "To increase translational biomedical research and improve the health of the people of Indiana and beyond." To this end, we have developed new mechanisms to acceleratetranslational research, including enhanced educational programs to train translational researchers,a newly designed community engagement activity to produce effective and bidirectional community partnerships, streamlining of all available research infra- structure to acceleratetranslational projects,and partnering with commercial and philanthropic organizations in Indiana. Throughout all of these efforts runs the critical link of our medical informatics program, enabling all parties to interact in a facile and responsive and prompt manner. Thus, the Indiana CTSI brings together the resourcesof an entire state. In short, it will provide to the national network of CTSAs: A true statewide laboratory to experiment with innovative methods aimed at transforming researchin biomedical sciences, health economy, health care delivery, and health policy. The 5 Specific goals of the Indiana CTSI: I. Create Translational Research Acceleration Programsand Support Pilot Projects by providing investigators and consumers with strategic leadership and mentorship to identify, evaluate, and support innovative and important pilot researchat each step of the translational cycle. . Train a new cadre of Translational Researchersby strengthening existing programs and creating new ones to educate trainees and engage faculty in the translational sciences. HI. Foster Robust Community Engagement by creating novel programs with bidirectional participation (i.e., from academia to the community and back again), such as the Indiana Community Health Enhancement Program (CHEP) and pilot programs in Health Econmics, and Rural and Global Health. IV. Build Facile and Comprehensive Research Resourcesand Technologies by transforming the existing and new research infrastructure into innovative programssuch as the Participant and Clinical Interaction Resources (PCIR), Translational Technology Resources(TTR). Research Ethics, Biostatistics and Design Program (BDP), and uniquely, biomedical engineering and bio-nanotechnology to facilitate health research. V. Leverage the Resources of the Greater Indiana Community by connecting to a broad array of resources from multiple partner institutions throughout the state to realize the full potential of the Indiana CTSI.

Two new training programs will be part of the Indiana CTSI. The K-12 program will fund post-doctoral junior faculty scientists, and the T-32 program will fund pre-doctoral research trainees. More specifically, the K12 program will focus on junior faculty, typically at the Assistant Professor level, who have already obtained a clinical doctoral degree (e.g., M.D., D.D.S., Pharm.D., O.D., D.P.T., Ph.D. clinical psychologist, etc.) and who desire to pursue clinical or translational research. The T32 program will focus on two types of pre-doctoral students: a) those enrolled in a clinical doctoral degree program as described above (i.e., students enrolled in medical, dental, pharmacy, optometry or other professional schools that provide a clinical doctoral degree);b) those enrolled in a Ph.D. program relevant to clinical or translational research (e.g., nursing or other health care professionals working on a Ph.D. or basic, social or other types of scientists focusing on translational research. These CTSI-funded training positions will not supplant the numerous T32 and individual K and other career awards currently funded at our institution, which reside in specific disciplines. Rather, the CTSI positions will serve a unique role in advancing multidisciplinary translational research. The overarching objectives common to both the K12 and T32 positions are: 1) Each CTSI-funded T32 or K12 trainee must focus on an area of translational research. 2) These CTSI positions will not be targeted to particular departments or diseases but will instead concentrate on multidisciplinary research involving a minimum of two different disciplines. 3) Each trainee funded by one of these CTSI positions will require co-mentorship involving a clinician scientist mentor and a non-clinician Ph.D. (or equivalent) mentor. 4) The degree or minor expected of a trainee funded by one of these CTSI positions will be an M.S. in Clinical Research or an M.S. in Translational Research (if a clinician-scientist) or a Minor in Clinical Research or a Minor in Translational Research (if a non-clinician pre-doctoral student). 5) Our goal is that a minimum of 25% of the CTSI-funded T32 and K12 positions will be occupied by trainees from under-represented minority groups. 6) There will be enhanced instruction in the responsible conduct of research (Bioethics Program, Section 15).

No abstract provided

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This supplemental award provides funds to CTSA Consortium-supported research projects for research on outcome measures in clinical and translational child health in priority areas as determined by the Best Pharmaceuticals for Children Act (BPCA) program administered by NICHD.

DESCRIPTION (provided by applicant): Anxiety disorders are some of the most common emotional disorders, cause considerable morbidity, and represent a significant economic burden. At least three key influences appear to be involved in the etiology of anxiety disorders: a) constitutive genetic differences, b) adverse early life event-mediated epigenetic changes, and c) chronic stress-induced plasticity. Our hypothesis for the present proposal will focus on two types of severe anxiety disorders, panic disorder (PD) and social anxiety disorder (SAD). We propose to study two components of the 'serotonergic dorsal raphe nucleus (DRN)-extended amygdala anxiety network': 1) the DRN-basolateral amygdala (BLA) and 2) the DRN-bed nucleus of the stria terminalis (BNST) pathways, which may be critical components in the pathophysiology of PD and SAD, respectively. We will systematically elucidate the behavioral, cellular and molecular effects of a) repeated stress neurotransmitter corticotropin- releasing factor (CRF)-induced plasticity, and b) two forms of genetic vulnerability, specifically, variations in serotonin transporter (SERT) expression using SERT and rats, and locally silencing the 5HT1A receptor gene in the BLA and BNST in the pathophysiology of PD and SAD in the current proposal. Our future studies will address the three-way interactions of stress plasticity and genetic vulnerability with early life stress (maternal separation)-induced epigenetic effects. In SPECIFIC AIM I, we will test the role of repeated stress- induced long-term plasticity in the BLA-DRN circuit in PD. In SPECIFIC AIM II, the role of long-term plasticity in the BNST-DRN circuit and SAD will be studied. SPECIFIC AIM III will determine the effects of reduced expression of SERT and vulnerability to stress-induced plasticity, whereas SPECIFIC AIM IV will study the role of reduced 5HT1A receptor gene expression in the vulnerability to stress-induced plasticity and pathophysiology of PD and SAD. Thus, the work proposed in this competitive renewal will a) characterize 'stress' and 'gene' interaction models of human anxiety disorders such as PD and SAD, b) study the putative disruption in the 'anxiety' circuitry of the BLA and BNST involved in the transition from adaptive panic and social responses to pathological states, and c) elucidate the molecular changes within the BLA and BNST that occur in anxiety disorders. The results of this project could help develop novel genetically targeted treatments and prevention strategies for disabling illnesses such as PD and SAD.

The Indiana Clinical and Translational Sciences Institute (CTSI) was created in 2008 at the crest of a wave of transformation of the three research universities in Indiana and has become a flagship catalyst for the state's biomedical Initiatives. The mission of the Indiana CTSI is to be an integrated home for clinical and translational research that creates a shared, investigator-friendly Institutional environment and provides easily accessible resources, services, and training to conduct the highest-quality research dedicated to improving human health across Indiana and beyond. The Four Specific Aims of the Indiana CTSI are: Aim I. Implement an Indiana roadmap for building the ideal research environment and collaborative platforms to conduct innovative research using our excellent multi-institutional, public-private partnerships throughout the state of Indiana and beyond by bringing together all three major research universities, Indiana, Purdue and Notre Dame, the state's key health care provider partners, private sector and corporate entities, and local and state agencies to create the infrastructure. Aim II. Further develop our successful Indiana CTSI established with the previous award into an integrated home that supports the careers and research projects of translational scientists and trainees through a streamlined structure, empowered leadership and administration, coordinated partner institution resources,.robust informatics infrastructure, innovative project development teams, and continuous evaluation and process improvement Aim III. Promote utilization of our innovative services and a well-tested network of resources and technologies in order to catalyze high-quality, efficient, and safe clinical and translational research. We have created Project Assistance and Resources for Translational Scientists (PARTS) suite of both established and new research support services and resources, a statewide network of technology cores, and a coordinated human subject research services and facilities. Aim IV. Extend our success in statewide education and training of clinical and translational research workforce through our Career development. Education and Research Training (CERT) suite of programs that includes successful existing programs, new training programs, and novel educational tools. RELEVANCE (See instructions): The Indiana Clinical and Translational Sciences Institute (CTSI) was created in 2008 as a statewide institute by the partnership of three research universities in Indiana (Indiana, Purdue and Notre Dame) and has become an essential resource for the state's biomedical initiatives. Here we describe its innovative vision, its impact, and its success in creating public-private partnerships to benefit the health and economy of the state.

Project Summary The present application Inhibition of nNOS-PSD95 interactions: A novel approach to developing treatments for PTSD addresses the critical need for efficacious treatments for posttraumatic stress disorder (PTSD). A key neural signaling cascade activated by a trauma experience is initiated by the excitatory neurotransmitter glutamate. Activation of the NMDA receptor, a glutamate receptor subtype, results in subsequent activation of the enzyme neuronal nitric oxide synthase (nNOS) and, ultimately, an increase in the production of the signaling molecule nitric oxide (NO). These events trigger aberrant synaptic plasticity that is implicated in the initiation and maintenance of PTSD. Postsynaptic density protein 95 (PSD95) targets nNOS to the NMDA receptor and is, therefore, required for NMDA receptor activation of nNOS. Our group first showed that the small molecule inhibitor IC87201 disrupts the functional protein-protein interaction between nNOS and PSD95 in vitro and attenuates NMDA receptor dependent hyperalgesia in vivo. We have now shown that IC87201 and a related analog, ZL006, block the long-term encoding of conditioned fear even after a fear conditioning session has occurred (i.e. post-trauma). Unlike NMDA receptor antagonists, these protein interaction inhibitors are efficacious without impairing motor movement or memory. Thus, disruption of signal compartmentalization represents an innovative approach to develop novel treatments for anxiety disorders with fewer side-effects. We have assembled a collaborative team to conduct work proposed under two Specific Aims. Aim 1 will test the benefits of inhibiting nNOS-PSD95 interactions in preclinical models of PTSD symptoms. Aim 2 will study the neural substrate mediating effects of nNOS-PSD95 protein-protein interaction inhibitors on conditioned fear. Results from this study will lay the foundation for preclinical development of nNOS targeting inhibitors as novel treatments for PTSD. These studies are expected to validate the disruption of signal protein compartmentalization as an innovative and feasible approach to drug development. The development of effective pharmacotherapies with novel chemical structures that possess limited side-effect profiles is expected to drive down escalating health care costs and alleviate unnecessary suffering in PTSD patients.

PROJECT SUMMARY The NCATS Human Subjects Research (HSR) Prior Approval (PA) process for KL2 Scholar Projects and certain Pilot Projects has been challenging for CTSA institutions to navigate. The Indiana CTSI will create a dedicated Quality Assurance/Quality Control (QA/QC) Project Manager role to improve quality and facilitate the HSRPA process. The long-term goal is to improve the quality of human subjects research submissions, make the overall process more efficient, and provide quality assurance for the ongoing conduct of research at the Indiana CTSI, with a particular emphasis on young investigators. The objectives are to decrease the review timeline from the time of award to study startup by supporting the education of young investigators and to ensure research conducted is at the highest quality so that important clinical research discoveries can positively impact patient outcomes more quickly. The specific aims are to: (1) Create a dedicated QA/QC position within the Indiana CTSI to perform quality reviews of CTSA-related submissions to NCATS, submit to the HSS, and manage the overall process; (2) Support applications by interfacing with KL2 Scholars and Pilot Project awardees throughout the IRB submission process; (3) Monitor funded projects once approved to ensure data quality and compliance; and (4) Offer QA/QC support to other K-funded young investigators. Accomplishing the above aims will enable measurable improvements in all stages of the HSRPA process and study conduct for KL2 Scholar Projects and certain Pilot Projects. In particular, individualized support to young investigators throughout their project will greatly enhance their training and preparation for a productive career in clinical research.