Steven Siegel - US grants

DESCRIPTION (provided by applicant): Studies in this Cutting-Edge Basic Research Award proposal would create a mouse model to assess genetic variability for in vivo physiological effects of NMDA antagonist drugs of abuse using auditory event related potentials (ERPs). Background: Abuse of NMDA receptor antagonists, such as ketamine and phencyclidine (PCP), has been recognized for many years. However, recent studies indicate that ketamine abuse has become more common among a subset of the population including people who frequent rave club parties, as well as military and medical personnel. The consequences of ketamine and PCP abuse include hallucinations, paranoia, disorganization and cognitive impairments. While many symptoms resolve within hours, others have been reported to last for many days. However, few studies have addressed genetic and other biological factors and that influence individual differences in ketamine and PCP sensitivity. Hypothesis: Animal studies with ketamine/PCP indicate that their behavioral effects and cellular toxicity are dependent on genetic background, suggesting differential vulnerability to their mechanism of action. Subanesthetic doses of ketamine/PCP have been proposed to exert their effect primarily by disrupting NMDA receptor-mediated glutamate transmission on GABAergic interneurons. This supports a hypothesis that genetic background may influence disruption of NMDA-mediated transmission on GABAergic interneurons to modulate the acute presentation and long-term sequelae of PCP/ketamine abuse. Research Project: The PI has developed a method to assess auditory ERPs in non-anesthetized mice with preliminary data demonstrating differential effects of ketamine among three inbred mouse strains. Aim 1 would determine the dose response relationship for the effects of acute ketamine administration on auditory ERPs in three inbred mouse strains. Aim 2 would then determine the differential sensitivity to effects of chronic ketamine on auditory ERPs in these three strains. Lastly, Aim 3 would determine the sensitivity to long-term changes following acute and chronic exposure to ketamine. Environment: The Stanley Center for Experimental Therapeutics in Psychiatry is a basic science laboratory within the Division of Neuropsychiatry and is part of The Center for Neurobiology and Behavior at the University of Pennsylvania. This laboratory contains all of the necessary resources to conduct the proposed studies of ketamine modulation of auditory ERPs in mice. Future Directions: Development of this model would facilitate examination of environmental, genetic and pharmacological factors that modulate the in vivo effects of ketamine and PCP. Such studies would also contribute basic knowledge regarding the neurobiological mechanisms of action, predisposition to toxicity and development of interventions directed at prevention of long-term sequelae following ketamine and PCP exposure.

[unreadable] DESCRIPTION (provided by applicant): Background: Medication nonadherence is the highest determinant of relapse and rehospitalization in schizophrenia. Therefore, interventions that empower patients to remain on medication for extended periods would substantially improve clinical outcomes using existing medications. The investigators previously developed a proof of concept long-term implantable delivery system using the typical antipsychotic agent haloperidol. Need: However, feedback from patients, families and physicians indicates that a long-term delivery system using a newer atypical antipsychotic medication would be preferable. Research Project: In response to previous reviews, the current proposal is now limited to 2 years to demonstrate an in vitro/in vivo correlation model for sterilized biodegradable Risperidone implants before proceeding to more costly toxicological, behavioral and molecular characterization studies. Studies were designed in consultation with the FDA Psychopharmacology team and would be performed in collaboration with the NIMH drug synthesis program. The first Aim will demonstrate in vitro release characteristics for a prototype antipsychotic medication from sterile implants made from 8 polymers at a single drug load. The second Aim will determine in vivo Risperidone serum concentration from each of these single-polymer implants. Dr. Louise Dube has joined our team to bring expertise in Clinical Psychopharmacology for ADME modeling. Environment: Studies will be conducted at the Center for Experimental Therapeutics in Psychiatry at the University of Pennsylvania under the direction of Steven J. Siegel MD, PhD. This program has a longstanding collaboration with Dr. Karen I. Winey in the Department of Material Science at The University of Pennsylvania. Investigators now have the required expertise and background in clinical psychiatry, Psychopharmacology and polymer engineering to accomplish the proposed research program. Industry R&D and pharmacokinetic modeling consultants have been included to further augment the research team. Future studies would evaluate the toxicological effects of a long-term subcutaneous delivery system as well as assessing molecular, neurochemical and behavioral measures to thoroughly evaluate long-term effects of Risperidone implants. Completion of the proposed studies would foster future industrial investment in long-term delivery systems for multiple agents, leading to dramatically improved patient care in the future. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): This is the second submission of a Phase II Cutting Edge Basic Research Award (CEBRA) that follows the Principal Investigator's Phase I application entitled, Evoked potentials and vulnerability to ketamine in mice (5-R21-DA-017082-02). All of the reviewers' concerns have been fully addressed in this revised application. Rationale: Experimentation with drugs of abuse is common among adolescents and young adults. Although the immediate consequences of intoxication are known, the ubiquitous nature of casual use leads many clinicians to discount mild drug abuse during high school and college as causal of later psychiatric symptoms and syndromes. Despite such assumptions, the lasting consequences of drugs that are emerging on campuses and in clubs across the nation, such as ketamine, are not known. During the Phase I portion of this study, the PI demonstrated persistent changes in Event Related Potentials (ERPs) following chronic ketamine exposure in adult mice. Preliminary data and previous literature also demonstrate that ketamine causes cellular pathology in brain. Questions that will be addressed: These findings lead to the following two overarching questions: 1) Does intermittent exposure to NMDA receptor antagonists during adolescence cause persistent physiological and cognitive deficits in adults? 2) Can adolescent ketamine abuse cause persistent alterations in cellular constituents and/or receptor signaling mechanisms in adults? Approach and Methods: Therefore, proposed studies will determine the duration of persistent ERP and cognitive deficits following ketamine exposure during adolescence in mice (Aim 1). Complementary studies will determine which cell classes are selectively altered by ketamine using stereological population estimates in adult animals following immunohistochemistry for cell-specific markers. Additionally, we will analyze measures of immediate cellular pathology directly following ketamine exposure during adolescence (Aim 2). Aim 3 will then utilize ex-vivo studies to determine the pattern of functional alterations of intracellular mechanisms of NMDA receptor mediated signal transduction among surviving cell populations following developmental exposure to ketamine. Significance: These three parallel Aims will address the degree of physiological, anatomical and biochemical functional neuroplasticity that exits following adolescent ketamine exposure. Therefore, completion of the proposed body of work will provide valuable evidence regarding the long-term, possibly irreversible, consequences of intermittent drug abuse during adolescence and early adulthood. Furthermore, these studies will address the potential roles of early drug abuse in later cognitive and psychiatric disorders throughout life.Although ketamine (Special K) abuse is common among adolescents, the consequences of this behavior are not known. To address this issue, we previously demonstrated that ketamine causes short-term changes in brain activity and signs of brain damage in adult mice. Proposed studies will determine the duration of abnormal brain function and the extent of brain damage following ketamine in adolescent mice to assess the potential for lasting, irreversible consequences of drug abuse during adolescence.

Rational: Social deficits are disabling, treatment refractory symptoms of schizophrenia. The amygdala is thought to modulate this behavior, and disruption of NMDA receptor (NMDAR) mediated glutamate transmission has been implicated as well. However, a detailed understanding of the cellular and regional circuit mechanisms underlying social deficits is lacking. Hypotheses: We propose that disrupted development and functioning of glutamatergic inputs to NMDARs on basolateral amygdala (BLA) neurons can disrupt acquisition and maintenance of normal social behavior. Furthermore, increased resting activity, i.e. noise, in BLA leads to disruption of normal signal processing and reduced signal-to-noise ratio (SNR) for social inputs from cortico-limbic brain regions. Approach: We will use in vivo electroencephalography, local field potentials (LFP) and multiunit recording in in BLA and hippocampus during social behavior as well as voltage sensitive dye imaging (VSDI) and intracellular recordings in slices from mice with disrupted NMDAR signaling. Model systems will include mice with constitutive reduction in NMDAR1 expression (NRl-/-) that have deficits in EEG and social interactions, as well as mice with amygdala-selective reduction in NRl using NRIflox mice with AAV-Cre injections. Interpretation: Data will inform interpretation of regional brain activation using fMRI and surface EEG (Project 1) in schizophrenia and at risk subjects. We will also examine novel pharmacologic approaches for restoration of excitatory-inhibitory balance in BLA at rest and during social behaviors. Public Welfare Statement: People with schizophrenia have difficulty in social interactions, which is disabling and resistant to current treatments. A part of the brain called the amygdala is thought to modulate normal social interactions, and disruption of the neurotransmitter glutamate at a receptor called the NMDA receptor, has been implicated in causing problems with normal social interactions in schizophrenia. This project will determine if disruption of glutamate activity at NMDA receptors in amygdala in mice can cause social deficits, and if fixing that activity with new medications could restore normal social function .

DESCRIPTION (provided by applicant): This application is for an individual fellowship for an MD-PhD student, with a research training plan designed to aid his long-term goal of becoming an independent physician-scientist in the field of innate immunity and inflammation. The inflammatory response is particularly important and carefully regulated in the respiratory tract, which must respond to a wide range of microbial insults while also preserving the gas-exchange function of the lungs. Bacterial colonization is rapidly countered by an inflammatory response, which includes the rapid release of mucins. Secreted mucins are heavily decorated with sialic acid, a carbon source that some bacteria, including the important respiratory pathogen pneumococcus, can catabolize. Preliminary data show that inflammation promotes pneumococcal growth in mice, and that the inflammatory response to bacterial colonization includes mucin secretion. The central hypothesis of this application is that airway inflammation promotes bacterial growth by providing sialylated mucin as a nutrient source. In Aim 1, a murine model of colonization will be used to determine the host and bacterial factors that contribute to inflammatory mucin secretion in the upper respiratory tract. Using a novel flow cytometric assay, we will determine whether this inflammatory response can promote bacterial growth in vivo, and whether that growth is dependent on sialylated mucin secretion. In Aim 2, we will determine whether influenza infection promotes sialylated mucin secretion, and define the innate signaling pathways required for this inflammatory response. We will determine whether sialic acid generated by influenza-induced inflammation promotes bacterial growth. These experiments will determine how the innate inflammatory response to pneumococcal colonization (Aim 1) and viral infection (Aim 2) promotes bacterial growth by providing a nutrient source through mucin secretion.

? DESCRIPTION (provided by applicant): The N-methyl-D-aspartate (NMDA) receptor hypofunction hypothesis is one of the leading postulates for the pathophysiology of schizophrenia (SCZ) and is supported by numerous pharmacologic, behavioral and genetic studies. Nevertheless, we have little insight into specific alterations in NMDAR signaling and its mechanistic basis in SCZ patients. This is a critical knowledge gap, which has impeded further development of this hypothesis and limited our efforts to identify specific therapeutic interventions. (Preliminary Data) As direct evidence for altered NMDA receptor (NMDAR) signaling, we found decreased NMDA/Glycine induced tyrosine phosphorylation of NMDAR subunit 2 (GluN2) and reduced downstream signaling in the postmortem dorsal lateral prefrontal cortex (DLPFC) of SCZ cases. These changes are not associated with decreased NMDARs but with reduced activity of a cascade of kinases- Src kinase, protein kinase C and Pyk2- which in concert decrease GluN2 tyrosine phosphorylation. We found multiple molecular alterations in the DLPFC of SCZ cases; increased PSD-95, increased erbB4 activity, decreased dysbindin -1 and RPTPa, each of which can induce Src hypoactivity. (Hypotheses) We hypothesize that hypoactivity of Src in the NMDAR complex (Src-NR) reduces GluN tyrosine phosphorylation and is caused by altered protein interactions in a network of Src-NR-associated proteins ( the Src-NR interactome), which can be leveraged to modify behavioral phenotypes of NMDAR hypoactivity. (Approach) We propose a human-rodent translation strategy, by which we analyze disease related alterations in postmortem brains and examine their underlying mechanisms in rodent studies. Aim 1 will further examine postmortem brains of an elderly and mid-life SCZ cohorts to identify molecular alterations in the Src-NR interactome in SCZ, Aim 2 will determine the role of protein interactions in Src-NR hypoactivity and test rescue strategies in ex vivo preparations of rodent and human postmortem tissues and Aim 3 will determine SCZ related behavior and EEG phenotypes of Src-/- mice and test if Src enhancement can rescue such phenotypes in vivo.

Contact PD/PI: Buchanan, Thomas A Inst-Career-Dev-001 (047) MCD PROJECT SUMMARY/ABSTRACT We propose a two-year Mentored Career Development (MCD) Program for clinician scholars that will lead to general competency in clinical and translational research and establish a lifelong methodological foundation overlaid with the additional competencies in communications, grant-writing, leadership and team science that are requisite for success as an independent career investigator. Consistent with the theme of the Southern California Clinical and Translational Science Institute (SC CTSI), the MCD Scholars will also develop the specific competencies and insights needed to carry out research in and translate findings to diverse populations. The MCD Program builds on a formal methodological core and complementary career development activities that have been established during the first four years of funding. We plan every-other- year cohorts of seven trainees, three supported by this award and four with institutional support. Trainees will enroll either in the existing Master of Science or Certificate in Clinical, Translational and Biomedical Investigations, completing a three-course methodological core, courses in biostatistics, and new courses on research in diverse populations and informatics. A major outcome for the core courses is completion of a K or R type proposal. The program for each MCD Scholar is tailored around the Individual Development Plan (IDP) and discussion with the mentoring team. Complementary activities include a career development seminar series, NIH-style mock review of planned applications, and mentor training. Scholars will have community- based experiences related to clinical and translational research and work with mentors from a community setting. They will have an array of internal and external opportunities to gain experience in regulatory science, in innovation and technology transfer, in health services and health outcomes, and in industry settings. A robust distance education platform is in place, which will be used for flexibility, archiving, and dissemination of materials, and training is offered in ?digital scholarship.? We intend to collaborate with the other California hubs to develop and disseminate a model curriculum in research on diverse populations, along with other products. Evaluation covers short-, intermediate- and long-term metrics, and the resulting data are used for continuing refinement of our program. The Multiple Principal Investigators, Drs. Samet and Patino-Sutton, have collaborated in developing the program over the first funding cycle; they are joined by faculty and mentors who bring expertise and experience with all phases of clinical and translational research and have rich records of research in diverse populations. Looking back over accomplishments since the SC CTSI was funded by the National Institutes of Health, we can identify transformative and widespread institutional change consequent to the current KL2 and TL1 programs. We have established a novel model for training at an institution that had only begun to formalize training in clinical and translational research before the SC CTSI programs were initiated. In this next phase, we will refine our training approach and aim to train researchers who carry out meaningful research to improve the health of diverse populations. Project Summary/Abstract Page 676 Contact PD/PI: Buchanan, Thomas A Inst-Career-Dev-001 (047) BLANK DOCUMENT Per the RFA-TR-14-009 instructions, Bibliography & References Cited is required for the Overall component, and not required for Administrative Core, Translational Science Base, Research Expertise and Methods, Research Implementation and Participation, Network Resources and Optional Modules, and Mentored Career Development components. References Cited Page 677 Contact PD/PI: Buchanan, Thomas A Inst-Career-Dev-001 (047) OMB Number: 4040-0001 Expiration Date: 06/30/2016 RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE - Project Director/Principal Investigator Prefix: First Name*: Jonathan Middle Name M. Last Name*: Samet Suffix: Position/Title*: Department Chair Organization Name*: University of Southern California Department: Preventive Medicine Division: Street1*: 2001 Soto Street Street2: SSB 330A City*: Los Angeles County: State*: CA: California Province: Country*: USA: UNITED STATES Zip / Postal Code*: 900899239 Phone Number*: 323.865.0803 Fax Number: 323.865.0854 E-Mail*: jsamet@usc.edu Credential, e.g., agency login: jsamet1 Project Role*: Other (Specify) Other Project Role Category: Project Lead, KL2 Director Degree Type: MD,MS Degree Year: File Name Mime Type Attach Biographical Sketch*: J._SAMET_-_BIOSKETCH.pdf application/pdf Attach Current & Pending Support: J._SAMET_- application/pdf _OTHER_SUPPORT.pdf Page 678 Tracking Number: GRANT11812224 Funding Opportunity Number: RFA-TR-14-009 . Received Date: 2015-01-14T19:51:42.000-05:00