Aaron Janowsky, Ph.D. - US grants

Tardive dyskinesia is a condition produced by long-term antipsychotic drug use, and affects millions of people world-wide. Though the problems has been termed a "dopamine supersensitivity syndrome," this undoubtedly describes only part of the underlying pathophysiology. Recent clinical evidence, in fact, suggests that calcium channel inhibitors may be useful for reducing the symptoms of tardive dyskinesia, as well as the symptoms of some other neuropsychiatric disorders. The goal of this proposal, therefore, is to further the understanding of dopamine and calcium related processes in animal models of human neuro- and psychopathology. Within a three year period, the experiments described in this proposal will examine the hypothesis that calcium channel inhibitors exert direct effects on dopamine receptor regulation and (or) function. Thus, this project will characterize: (1) the effects of chronic administration of calcium channel inhibitors on the development and the maintenance of neuroleptic induced changes in dopamine receptor characteristics and activity, (2) the interaction in vitro between calcium channel inhibitors, adenylate cyclase activity, and dopamine receptor characteristics, and (3) the role of dopamine (lesions) in the effects of calcium channel inhibitors on dopamine receptor regulation and function. The experiments will characterize i) changes in D1 ((3H)SCH23390) and D2 ((3H)spiroperidol) receptor recognition site density and affinity for ligand, ii) agonist displacement of the respective radioligand in the presence and absence of guanine nucleotide and iii) changes in D1 receptor mediated activation and D2 receptor mediated inhibition of adenylate cyclase activity in rat brain striatum.

Alcoholism is a clinical problem that affects millions of people world-wide. Although various aspects of the disease have been extensively studied, its molecular and biochemical etiologies are not understood. Recently, new tools have become available for use in characterizing the symptomatology of alcoholism, and some potential predisposing factors that may play a role in the development of the disease and (or) its clinical course. Mouse lines have been developed for their susceptibility to seizures during withdrawal from chronic ethanol exposure. Thus, withdrawal seizure prone (WSP) and resistant (WSR) mice provide a behavioral model for use in characterizing physiological changes that accompany physical dependence on alcohol. In addition, molecular and biochemical mechanisms that predispose WSP mice to symptoms of withdrawal, and protect WSR mice, can be examined using new radioligands that bind to sites of anticonvulsant action. Within a three year period, the experiments described in this proposal will examine the hypothesis that alterations in the excitatory amino acid receptor - linked ion channel complex in brain precede and accompany susceptibility to seizures during withdrawal from ethanol. Thus, this project will characterize: (1) the sensitivity of [3H]glutamate receptors to glycine and the sensitivity to glutamate and glycine of the associated ion channel binding site for radiolabelled MK-801, a potent and selective anticonvulsant agent, in brains from WSP and WSR mice. (2) the effects of chronic ethanol exposure on each of the above interactions in brain tissue from WSP and WSR mice, and (3) the effects of withdrawal from ethanol on the interactions among receptor sites in the glutamic acid receptor linked - ion channel complex in brain tissue from WSP and WSR mice. The results of these studies could suggest biochemical mechanisms that are involved in the pathophysiology of symptoms of withdrawal, and indicate new directions in research for the development of specific pharmacotherapies that can be used in the treatment of those symptoms.

The National Institute on Drug Abuse (NIDA), Medications Development Division (MDD) has established a Cocaine Treatment Discovery Program (CTDP) to accelerate the identification of potential medications for the medical management of cocaine dependence. This program involves testing compounds through a decision-based screening scheme designed to identify potential treatment agents which will either antagonize the effects of cocaine or substitute for cocaine. The purpose of this contract is to determine the affinity of up to 600 test compounds at biogenic amine transporter (dopamine, serotonin, and norepinephrine) binding sites. In addition, up to 500 of these compounds will be assessed for their ability to inhibit the reuptake of biogenic amines and up to 200 of them will be evaluated for their ability to facilitate an "amphetamine-like" release of biogenic amines. Compounds will, in general, be submitted by NIDA to the contractor "blind" with a coded identification number and information about the compound such as their physical properties and solubility. CTDP data generated from this contract on proprietary compounds will be maintained in confidence by NIDA for three (3) years from the date of reporting of each test result to the compound submitter. After three (3) years, NIDA will disclose this information into the public domain through its MDD structure-activity database. Compound submitters are free to disclose CTDP data generated on their compounds prior to the three (3) year period of non-disclosure. The contractor will meet annually with NIDA and consultants with expertise in in vitro biogenic amine transporter assays to review the quality of the contract data and discuss new state-of-the-art technologies and methodologies for potential incorporation into the contract.

The Administrative Core (Component #1) of the Methamphetamine Abuse research Center (MARC) contains the centralized administrative functions for all 3 Scientific Components (#7-9), as well as for all of the support cores (#1-6), including the Biostatistics (#2), Animal (#3), Education (#4), Translational Service (#5), and Pilot Projects (#6) Cores. The Administrative Core coordinates the scientific oversight provided by the Scientific Advisory Board, including review of Pilot Projects, and educational enrichment as provided by such activities as workshops. Executive decisions regarding MARC activities will be made by the MARC Executive Committee of Scientific and Core Component Pis. The Executive Committee will continuously evaluate progress toward general scientific research goals, and will oversee quality control mechanisms in all areas of Center activity. Additionally, administrative support will be provided for animal production (selection), genetics, statistical analyses both within and across components, educational enrichment and training, human and animal imaging studies and tissue work. Oversight will be provided for centralized core facilities characterizing and assaying human and animal samples. It will oversee genetics, bioinformatics and data sharing, biostatistical analyses, and training and educational enrichment and outreach activities. Finally, the Administrative Core will coordinate Pilot Project solicitation, review and award. Thus, the Administrative Core is structured to address two specific aims. Specific Aim 1: The Core provides administrative and budgetary oversight and the general coordination of all Center activities, and Specific Aim 2: The Core coordinates communication among Center investigators via a secured shared drive on the University network, and the dissemination of information to outside entities, including other investigators and Centers, via the MARC web site. The general administrative coordination of all Center activities and the Center's budget will be supervised by Dr. Janowsky, the Center Director. The coordination of and future planning for scientific directions will be supervised by Dr. Phillips, the Scientific Director, who will act as Director in Dr. Janowsky's absence.

DESCRIPTION (provided by applicant): Oregon is one of a few states where methamphetamine (MA) abusers without a greater history of cocaine abuse are recruited for clinical research. The Methamphetamine Abuse Research Center (MARC) will characterize effects of MA administration and withdrawal at molecular, neurochemical, anatomical, behavioral, and clinical levels, to identify obstacles to recovery in MA abusers. Overarching themes addressed in preclinical and clinical research components using common methodologies include neuroadaptation to MA, neuroanatomy of MA-related behaviors, stressor responsivity, and impulsivity. Human and animal protocols inform one another with respect to these themes, forming the basis for bidirectional translational research. The Center's Administrative Core facilitates interactions among MARC investigators, and the Biostatistics Core analyzes associations and correlations within and across components. The Animal Core breeds and maintains genetically selected lines and other strains, and provides them to MARC investigators. The Education Core coordinates the research training of M.D. and Ph.D. pre- and postdoctoral fellows, and organizes and disseminates both clinical and preclinical information from MARC investigators to Centers and Oregon's rural areas that are impacted by MA production and abuse. Scientific Component 5 correlates acetylcholine release with active and passive administration of MA and provides neuroanatomic and molecular signaling data relevant to MA seeking for use in other components. Scientific Component 6, examines genetic influences, and behavioral and neurochemical correlates (in collaboration with Component 5) in mice selected for differences in MA-induced neuroadaptation and MA self-administration. Nomination of genetic candidates from this preclinical work could inform treatment strategies in clinical Component 7, and specific genetic associations can be pursued in patients from clinical components. Scientific Component 7 evaluates the relationship between symptom severity at intake and treatment failure in inpatients, tests (3-receptor and cholinergic medications, and conducts human laboratory evaluations of stressor responsivity. Component 8, the Pilot Projects, provides a means to support the development of multiple new directions in research on MA abuse. Thus, the MARC's themes address clinically relevant issues using innovative multidisciplinary approaches.

Oregon is one of a few states where methamphetamine (MA) abusers without a greater history of cocaine abuse are recruited for clinical research. This renewal of the Methamphetamine Abuse Research Center (MARC) will characterize effects of MA at molecular, genetic, neurochemical, anatomical, behavioral, and clinical levels, to identify risks and obstacles to recovery in MA abusers. Integrated preclinical and clinical research components use some common methodologies and address MA-related themes of neuroadaptation, neurocircuitry, and neuroimmune effects. This renewal continues to pursue bidirectional translational research in which human and animal results inform one another. The Center's Administrative Core 1 supervises budgetary issues and facilitates interactions among MARC investigators, and the Biostatistics and Genetics Core 2 provides statistical and genetic analysis services. The Animal Core 3 provides genetic animal models and some behavioral testing services to MARC investigators. The Education Core 4 coordinates the research training of M.D. and Ph.D. pre- and post-doctoral fellows, and disseminates clinical and preclinical information from MARC investigators to other Centers and more rural areas that are impacted by MA abuse. The Translational Service Core 5 recruits and characterizes subjects and conducts identical biochemical assays on human and MA drinking selected mouse line samples. The Pilot Projects Core 6 supports the development of multiple new directions in research on MA abuse. Scientific Component 7 associates image analysis results with impulsive decision making in human and animal subjects. Scientific Component 8 examines the role of immune function in human cognitive response and tests a novel immunotherapy in mice, and Scientific Component 9 uses MA drinking selected mouse lines to examine the role of immune function in the risk for MA self-administration and how this risk interacts with MA effects on immune function. Data and samples are shared and compared across components. Thus, the MARC addresses clinically relevant themes using integrated, innovative, multidisciplinary, and translational approaches.