Charles B. Nemeroff - US grants

Neurotensin (NT), an endogenous peptide, exerts many actions similar to those of neuroleptic drugs. Substantial progress by our group, under support of this grant, and by other groups shows that NT modulated dopaminergic transmission in the mesolimbic system, as assessed by several neurochemical and pharmacobehavioral techniques. Interactions with dopmaine (DA) systems are of special interest because at least four important disorders of the central nervous system appear to be related to abberations of DA metabolism, schizophrenia, Parkinson's disease, Huntington's chorea and tardive dyskinesia. We now propose to study autopsy material from patients who in life suffered from one of these conditions. In addition, NT will be measured in cerebrospinal fluid (CSF) from schizophrenic patients and appropriate controls. The studies pertaining to schizophrenic tissue and CSF are intended to test the reproducibility of present findings that in such patients NT levels are elevated in frontal cortex, while in a subset of schizophrenics, NT is reduced in CSF. The neurotensin receptor binding studies in post-mortem tissue represent a novel research direction. Animal studies are designed further to explicate the exact role of NT in neurophysiological transactions. We will examine the possible mediation of NT effects by transmitters other than DA, possible actions of NT on DA autoreceptors, possible interaction of NT with the mesocortical DA system, the activity of NT in animal models of tardive dyskinesia, the possibility that chronic NT treatment may produce mesolimbic DA supersensitivity, and cognate issues. Both human and animal studies depend, in part, on the availability in our laboratory of a sensitive and specific NT radioimmunoassay, an NT receptor binding technique and HPLC technique with electrochemical detection for catecholamine assay and a variety of behavioral techniques. The long-term objective of this project is to add to our fund of knowledge regarding the pathophysiology of certain diseases of the human nervous system and thus to improve current management and treatment.

The neurochemical pathology of Alzheimer's disease (AD), the most common cause of dementia, has received increasingly more attention. It is now evident that specific chemically-defined neurotransmitter systems are pathologically involved in this neurodegenerative disorder. Neurons affected include cholinergic neurons in the basal forebrain, GABA-containing neurons and two groups of neuropeptide-containing cells: somatostatin- and CRF-containing cells in the cerebral cortex. Other peptidergic neurons such as those containing cholecystokinin and vasoactive-intestinal peptide are apparently spared. In the present competitive renewal application we seek to continue to scrutinize in detail, the dynamic state of cholinergic neurons using tissue obtained in the Rapid Autopsy Procedure (20-60 min after death). By measuring markers of neuronal integrity, neuronal activity (high affinity choline uptake), the choline transporter itself, receptor number and affinity, and second messenger as well as third messenger responses (protein phosphorylation), the dynamic state of cholinergic neurotransmission will be assessed in several brain regions from histologically-confirmed AD and age- and sex-matched controls. In addition, we shall continue our work on peptidergic (SRIF and CRF) systems in Alzheimer's disease including assessment of the relationship between CSF and brain concentrations of these peptides. Moreover, SRIF receptor subtypes will be measured, as will functional responses of these receptors. We shall also determine the relationship between the presence of the abnormal ADassociated protein, Alz 68, and the alterations in ACh and peptidergic neurons. We shall also scrutinize alterations in GABA-containing neurons in AD. The majority of these studies are feasible because of the unique availability of the rapid autopsy tissue - our center is the only one in the world conducting this procedure. These studies will provide novel data that may well result in the development of a rational drug therapy in patients with AD.

Since elucidation of its structure in 1981, corticotropin-releasing factor (CRF), a neuropeptide comprised of 41 amino acids, has been shown to function as a hypothalamic releasing hormone by regulating the secretion of corticotropin (ACTH) and other pro- opiomelanocortin-related peptides from the anterior pituitary. The heterogeneous distribution of CRF and CRF receptors in hypothalamic and extrahypothalamic regions of the mammalian CNS, and its electrophysiological and behavioral effects, all are concordant with an extra-endocrine role for this peptide in higher brain centers. The present proposal seeks to utilize preclinical and clinical paradigms to provide further evidence for a role for CRF as a neurotransmitter in the CNS, and furthermore, to investigate the hypothesis that CRF is hypersecreted in patients with major depression. Towards the former goal, we shall elucidate CRF-containing neural circuits in the rat CNS by a combination of chemical lesioning techniques and use of a sensitive and specific radioimmunoassay for the peptide, determine what neurotransmitters/neuromodulators regulate CRF release, determine the subcellular distribution of CRF, determine whether cyclic nucleotides mediate CRF responses in the CNS and finally, determine the role of CRF neurons in responses to acute and chronic stress. Towards the latter goal, we shall attempt to confirm and extend our previous findings that cerebrospinal fluid (CSF) concentrations of CRF are elevated in patients with major depression by studying a large population of well-characterized, drug-free neuropsychiatric patients. The specificity of this finding will be studied by measurement of CSF CRF, not only in patients with major depression, but also in schizophrenic and demented patients, as well as patients with Huntington's chorea and multiple sclerosis. We shall determine whether the elevation in CSF CRF-L1 is state-dependent by: 1) determining whether treatment with electroconvulsive therapy (ECT) produces a significant reduction in CSF CRF-L1, and, 2) determining whether treatment with tricyclic antidepressants in multiple sclerosis patients with depression results in a reduction in CSF CRF-L1. Finally, the specificity of the blunted ACTH response to intravenously (IV)-administered CRF in depressed patients will be explored. We shall study patients not only with major depression, but also those with diagnoses of schizophrenia, post- traumatic stress disorder and panic disorder, resulting in novel information on the role of CRF in affective disorders.

DESCRIPTION: (Adapted from the Investigator's Abstract) There is considerable evidence neurons that utilize the neuropeptide neurotensin (NT) are involved in both the mechanism of action of antipsychotic drugs (APDs) and the pathophysiology of schizophrenia. Despite this burgeoning database, there is no direct evidence to date that NT neurotransmission mediates behaviors relevant to the clinical efficacy of APDs. Conclusive demonstration of this association is the overall objective of this proposal. Hypotheses to be tested include the following; 1) Do all APDs increase NT release in the nucleus accumbens? 2) Is this increase in NT neurotransmission sufficient and necessary for the effects of APDs in behaviors relevant to clinical efficacy? 3) Are sex differences in schizophrenia related to ovarian hormone regulation of the NT system? 4) Are there specific alterations of the NT system in postmortem tissue consistent with an involvement of this system in the pathophysiology of schizophrenia? Thus, specific aim #1 scrutinizes the role of NT neurotransmission in animal models predictive of the clinical efficacy of APDs. Specific aim #2 evaluates the putative involvement of NT neurotransmission in the therapeutically relevant behavioral effects of chronically administered APDs. Two behavioral paradigms (latent inhibition and prepulse inhibition (PPI) of the acoustic startle reflex) with face, predictive and construct validity as animal models of sensorimotor gating in schizophrenia will be utilized. Specific aim #3 examines the behavioral consequence (specifically on sensorimotor gating ) of deficient NT neurotransmission in transgenic mice lacking NT. Specific aim #4 would correlate ovarian hormone regulation of NT neurotransmission (including NT gene expression, NT release, and the electrophysiologic responsiveness of nucleus accumbens neurons to NT) with estrous cycle regulation of sensorimotor gating. Specific aim # 5 explores a novel mechanism by which NT may directly modulate the D2 dopamine receptor. Specific aim #6 continues examination of NT neuronal systems in postmortem brain tissue from patients with schizophrenia, bipolar depression, or normal controls, with focus given to brain regions implicated in sensorimotor gating. This linkage of studies is intended to provide the first definitive evidence that intact NT neurotransmission is critically involved in behaviors relevant to the clinical efficacy of APDs, and provide clear rationale for the use of NT agonists as novel APDs.

DESCRIPTION: (Adapted from applicant's abstract) This application is the competitive renewal application of NIH MH-42088, seeking five years of support, which would represent years 15-19 of this project on the Psychobiology of Corticotropin-Releasing Factor (CRF). In the past years, this grant has provided much evidence for a preeminent role for CRF-containing neurons in mediating the response, characterized in considerable detail the neurotransrnitter role of CRF in the mammalian CNS, and perhaps most importantly, contributed to the burgeoning dah base which supports the hypothesis that CRF is hypersecreted in patients with major depression. Indeed, contributions from this and other laboratories using a variety of multidisciplinary approaches have provided unequivocal evidence for hyperactivity of CRF-containing in neurons patients with major depression, particularly those patients with severe depression. The current proposal seeks to extend this work in directions largely based upon findings obtained during the current funding period (aims I & 2 in a new direction as well (genetics aims 3 & 4). Specifically, the investigators wish to continue the postmortem brain tissue studies to further characterize that alterations in CRF-containing neurons and CRF receptor-containing neurons by scrutinizing CRF and CRF receptor mRNA expression, as well as focusing on the CRF2 receptor and urocortin The development of small molecule, non-peptide CRF1 receptor antagonists has occurred to some extent due to the investigators progress in this field. With their increasing availability, the behavioral, endocrine, and neurochemical responses chronic administration of these CRF1 receptor antagonists will be characterized. Moreover, the responses to discontinuation of these compounds will be assessed. In a novel animal model of depression based upon sensitivity stress in the forced-swim test that is highly sensitive to all classes of antidepressants, the investigators will determine whether CRF1 receptor antagonists exhibit an antidepressant profile. Moreover, are alterations in CNS CRF neuronal systems and receptors present in this animal model? Because evidence suggests that altered CRF function appears to be responsible, at least in part, for the anxiety and/or depression phenotype in some animal models, the investigators will map the quantitative trait loci (QTL) that control the expression of the CRF system. These studies will provide novel information on the role of CRF nepronal systems in the pathophysiology of depression, in animal models of systemic CRF1 receptor antagonist administration.

Clinical lore, case reports and a few prospective studies all have provided data consistent with the hypothesis that there is an extraordinarily high rate of depressive symptoms in patients with pancreatic cancer. It is frequently stated that psychiatric morbidity may in fact precede any of the other signs and symptoms of this carcinoma. The goals of this study are: (1) to determine whether the prevalence of major depression and other psychiatric disorders is, in fact, increased in newly diagnosed cases of pancreatic cancer as compared to patients with esophageal or gastric cancer; (2) to determine, using well established biological markers of major depression [reduced density of H-imipramine platelet binding sites, non-suppression of cortisol to dexamethasone and increased cerebrospinal fluid concentrations of corticotropin-releasing factor (CRF) and thyrotropin-releasing hormone (TRH)], whether depressed patients with pancreatic cancer exhibit these alterations when compared to groups of normal controls, patients with major depression without cancer, patients with esophageal or gastric cancer and patients with pancreatic cancer without depression; (3) to determine, in view of recent studies suggesting that depressed patients may have impaired immune function, whether the depressed cancer patients we shall study exhibit alterations in immune function compared to non-depressed patients; and (4) to determine whether pancreatic carcinoma produces interleukin-1 and other lymphokines that may contribute to depression and pituitary-adrenal hyperactivity. These studies, taken together, will provide novel data concerning the prevalence and biology of depression in patients with pancreatic cancer.

The Department of Psychiatry and Behavioral Sciences of the Emory University School of Medicine is applying for a Research Infrastructure Program (RISP) Grant. Funds are requested for partial salary support for junior faculty, especially women and minorities, and training expenses for junior faculty which take the form of support of the following modules: (l) A Patient Recruitment and Assessment Module, which will provide standardized psychiatric diagnostic and dimensional rating scales, as well as recruitment of patients with different psychiatric diagnoses and age- and sex-matched normal controls; (2) A Neuroendocrinology/Neurochemistry/Brain Imaging Module, which will provide consultation for measurement of routine hormones and neurotransmitters (and metabolites), as well as measurements of transporters and receptors. In addition, measurement of the concentrations of centrally acting drugs, including antidepressants, anxiolytics and antipsychotics in plasma and other biological fluids is available. This module will also provide expertise in structural (MRI) and functional (PET, SPECT, MRS) brain imaging; (3) A Biological Tissues and Fluids Module, which will provide for cataloging and storage of plasma, urine and cerebrospinal fluid (CSF) samples, as well as platelets, lymphocytes and postmortem brain tissue from well-characterized patients diagnosed in life with a major psychiatric disorder; (4) An Experimental Design and Biostatistics Module, which will assist junior investigators with research design, as well as statistical analysis of the data generated. In order to maximize the likelihood of a rapid advancement in the quality and magnitude of clinical research in the Department, support is also requested for a modicum of inpatient bed costs at Emory University Hospital, travel support for junior faculty to attend national meetings, as well as for funding of an external Scientific Advisory Board to convene at Emory once per year to review the program. The support provided by the RISP, together with that provided to the Department by the Emory University School of Medicine (new laboratory and office space, seed money for new faculty, and support of senior faculty, as well as the new positron-emission tomography (PET) center), taken together with the more than ample patient population in the greater Atlanta area, will result in significant and lasting improvements in the Department's capacity to obtain extramural research funding.

In spite of a plethora of published studies in laboratory animals that have documented effects of cocaine on specific central nervous system neurotransmitter systems, there is remarkably little known about the chronic effects of this psychostimulant on the human brain. In this proposal we seek to scrutinize the effects of chronic cocaine abuse in humans by assessment of neurochemical alteration in approximately 50 brain regions derived from post-mortem brain tissue of confirmed cocaine abusers (with segmental hair analysis to document the temporal pattern of cocaine use) and age- and sex-matched controls. Through cocaine's actions on monoamine transporters, the preclinical literature has clearly implicated dopamine (DA), serotonin (5MT) and norepinephrine (NE)-containing neurons in mediating the actions of cocaine; there is also considerable evidence implicating two neuropeptide neuronal systems (corticotropin-releasing factor [CRF] and neurotensin [NT]) in the actions of this drug of abuse. Alterations in the monoamine transporters will be assessed by measurement of: radioligand binding, transporter protein concentration by the use of newly developed specific antibodies to the transporters, and mRNA expression of the transporters. Alterations in CRF and NT systems will be assessed by measurement of the neuropeptides, neuropeptide mRNA expression, neuropeptide receptor binding, neuropeptide receptor mRNA expression, and in the case of the CRF system, the CRF-binding protein and CRF signal transduction. These studies will provide novel data on the effects of cocaine abuse on specific neurotransmitter systems in the human central nervous system.

Alzheimer's Disease (AD), a neuropsychiatric disorder, is characterized by severe morbidity and is one of the leading causes of death in the elderly in the United States. It comprises more than 50% of the cases of dementia. Although considerable progress has been made in the genetics, neurochemical pathology, neuropsychology, and neuroanatomy of AD, little is available in the way of treatment, with the exception of the recent FDA approval of an acetylcholinesterase inhibitor, which is helpful only in a minority of patients. This revised competitive renewal application seeks to build upon our progress in characterizing the neurochemical pathology of neurons containing neuropeptides and other neurotransmitters in AD. The major focus of this application is intensive scrutiny of the nature of the pathological involvement of neurons containing corticotropin-releasing factor (CRF) in AD. Our group was the first to describe the marked reduction in CRF immunoreactivity in post-mortem brain tissue in patients with AD, and this finding has been widely confirmed. In the present application, we seek to scrutinize this finding more closely by measurement of several indices of CRF neuronal function in post-mortem brain tissue in AD and in controls. These measures include: CRF immunoreactivity, CRF receptor density and affinity, mRNA expression for both CRF and the CRF receptor, measurement of the concentration and expression of the newly discovered CRF binding protein, as well as measures of intracellular signal transduction in response to CRF. In addition, in view of the many reports of widespread serotonergic and noradrenergic neuronal dysfunction in AD, we shall utilize newly available antibodies to the 5-HT and NE transporters to determine the cellular localization and quantify the presynaptic 5-HT and NE transporter proteins in post-mortem brain samples. The resultant data will also provide novel information on the relationship between the pathological involvement of these three neurotransmitter systems in AD. Taken together, these experiments will further our understanding of the neurochemical pathology of AD and provide the basis for the development of novel therapies for this disorder.

Neurotensin (NT), an endogenous tridecapeptide, is closely associated with CNS dopamine (DA) systems within the mammalian central nervous system (CNS). When administered directly into the CNS of laboratory animals, NT produces pharmacological, neurochemical and behavioral effects similar to those of antipsychotic drugs, particularly those classified as atypical. Typical antipsychotic drugs such as haloperidol increase NT concentrations and proNT mRNA expression in both the nucleus accumbens (a terminal area of the mesolimbic DA system) and the caudate nucleus (a terminal area of the nigroneostriatal DA system). Atypical antipsychotic drugs such as clozapine increase NT concentration and proNT mRNA expression only in the nucleus accumbens. These and other data suggest that the increase in these measures of NT biosynthesis in the nucleus accumbens are predictive of antipsychotic efficacy, whereas the increase in the caudate nucleus are predictive of extrapyramidal side effect (EPS) liability. There is also evidence that NT neuronal function is altered in schizophrenia. The present competitive renewal application seeks to focus on effects of typical and atypical antipsychotic drugs on the synaptic availability of NT in the nucleus accumbens and caudate nucleus, as well as on NT receptor kinetics and NT receptor mRNA expression. The intracellular signalling pathways mediating antipsychotic drug effects on c-fos and NT gene expression will be explored in primary neuronal cultures. Ontogeny and gender effects of antipsychotic drugs on NT neurons and on the CNS effects of NT will be scrutinized, a study of particular interest, in view of increasing evidence that schizophrenia is a neurodevelopmental disorder, and that there are gender differences in this disorder. We will study whether the behavioral effects of typical and atypical antipsychotics are mediated through NT receptor mechanisms. The brain sites of action of intracerebroventricularly administered NT will be mapped by fos immunohistochemistry. The electrophysiological effects of NT itself, and its actions on DA in the major subdivisions of the nucleus accumbens will be characterized. Signal transduction mechanisms mediating the effects of NT receptor activation will be explored in primary neuronal cultures. Finally, a major focus of the present proposal is assessing the role of NT neuronal system alterations in the pathophysiology of schizophrenia by measurement of various indices of NT neuronal function (NT, NT mRNA expression, NT receptor binding and NT receptor mRNA expression) in post- mortem brain tissue of patients who in life were diagnosed as schizophrenic, age- and sex-matched controls and a unique control group (patients with bipolar disorder treated with antipsychotic drugs). These studies will provide novel information concerning the role of NT systems in the mechanism of action of antipsychotic drugs, and in the pathophysiology of schizophrenia.

tissue resource /registry; biomedical facility; human tissue; postmortem;

This study will evaluate potential alterations in platelet function and sympathetic tone in patients with major depression. Preliminary data were analyzed, and results were used to prepare a grant application.

This study is an ammendment to SPID 3250. It will evaluate alterations in platelet reactivity and sympathetic tone in patients with major depression, and then re-evaluate these patients after 6 weeks of treatment with the anti-depressant, paroxetine.

A preeminent role of early adverse experiences in the pathogenesis of psychiatric disorders has long been postulated and has been supported by recent studies. We hypothesize~that early adverse experiences may induce an increased sensitivity to the effects of stress later in life and render an individual vulnerable to stress-related psychiatric disorders. This vulnerability may be mediated by persistent changes in corticotropin- releasing factor (CRF)-containing neurons and the hypothalamic-pituitary- adrenal (HPA) axis as well as noradrenergic and serotonergic neurotransmitter systems. In this project, we will characterize neurobiological consequences of early adverse experiences, i.e. childhood sexual and/or physical abuse. Specifically, we will compare indices of central CRF activity, HPA axis function, noradrenergic and serotonergic neuronal activity as well as sympathetic-nervous system activity, and structural alterations in several brain regions in women with a personal history of childhood abuse (early life stress, ELS) and current major depression (ELS/MDD), women with a history of childhood abuse without major depression (ELS/non-MDD), women without a history of childhood abuse and major depression (non-ELS/MDD) and women without a history of childhood abuse and no psychiatric disorder (CONTROLS). Based on our findings in animal models, we also will evaluate whether the profound neuroendocrine consequences of early life stress are reversed "by treatment with the selective serotonin reuptake inhibitor, fluoxetine. Using structural MRI methods, the volume of the hippocampus will be compared in the four diagnostic groups. In collaboration with the functional brain imaging core, these subjects will participate in studies to measure CNS density of both the serotonin transporter and the CRF1 receptor, as well as fMRI measures of stress reactivity. Findings from this project will increase our understanding of the pathogenesis of psychiatric disorders and will help to develop new intervention strategies to prevent adulthood psychopathology in victims of childhood abuse.

This is the competitive renewal application for the Emory Conte Center for the Neurobiology of Mental Disorders (CCNMD) in the Department of Psychiatry and Behavioral Sciences (DPBS) at the Emory University School of Medicine. The goal of this research plan is to utilize state of the art methods ranging from molecular biology to cognitive neuroscience and functional brain imaging to characterize the neurobiological mechanisms by which adverse early life stress (ELS, eg., childhood sexual or physical abuse), increases an individuals' vulnerability to depression and related disorders. Our data suggests that the pathophysiology of major depression in patients with a history of ELS is distinct from that observed in patients with major depression without ELS. Such findings have major implications for nosology, neurochemical pathology and treatment of patients with these disorders. Supported by 4 cores (Administrative, Rat, Assay and Primate), nine projects seek to further characterize the long term neurobiological consequences of ELS in rodents, non-human primates and humans. Using rat and rhesus monkey models of ELS and patients with major depression or social anxiety disorder (SAD) with or without ELS, we focus on alterations in CNS neuroregulators 0001, Paul Plotsky, PhD, PI and immune system dysfunction 0009, Andrew Miller, MD, PI. Michael Davis, PhD, PI utilizes a novel model of learned safety to scrutinize our non-human primate and human patients to further elucidate the long-term behavioral consequences of ELS. The role of ELS in cognitive dysfunction of depression is the focus of 0010, in non-human primates (Stuart Zola, PhD, PI) and also an important component of 0007 (Clinical Psychobiology, Charles Nemeroff, MD, PhD, PI). The latter project also will determine whether men with depression and ELS exhibit the same neuroendocrine and autonomic alterations observed in women, whether patients with SAD and ELS exhibit similar or different neurobiological alterations as patients with major depression and ELS, and will better define the vulnerable periods in development and type of early ELS that contribute to the observed CNS alterations. 0012 (Helen Mayberg, MD and Gregory Berns, MD, PhD, PI) and 0013 (Clinton Kilts, PhD and Xiaoping Hu, PhD) use state-of-the-art functional imaging modalities (PET, fMRI) and diffusion tensor imaging to determine the contribution of ELS to the depression-associated alterations in regional CNS activity, as well as to assess alterations in structural and anatomical connectivity. Finally, 0011 (Zachary Stowe, MD, PI) utilizes a unique clinical population, maternal depression during pregnancy and in the postpartum period, as a novel paradigm to study the consequences of ELS in their offspring. The results of these studies will provide novel data that will likely impact upon pathophysiological subtyping of depression, understanding the neurobiology of risk factors, as well as translating into predictors of treatment response and novel treatment strategies.

This study is an amendment to SPID 3250. The purpose of this study is to evaluate alterations in platelet reactivity and sympathetic tone in patients with major depression and then to re-evaluate these patients after 6 weeks of randomized, double-blind treatment with the antidepressant, paroxetine, in comparison to treatment with placebo.

This application, in response to RFA: MH-03-008 (National Cooperative Drug Discovery Groups for the Treatment of Mood Disorders or Nicotine Addiction, NC 336-MD/NA), proposes the creation of"The Emory-GSK-NIMH Collaborative Mood Disorders Initiative." This unique opportunity to accererate antidepressant drug development brings together expertise of three complementary research groups: the Emory University School of Medicine Department of Psychiatry and Behavioral Sciences, the Mood and Anxiety Disorders Program at NIMH and the Center for Excellence in Drug Discovery (CEDD) in Psychiatry of GlaxoSmithKline (GSK), one of the largest multinational pharmaceutical companies. The two major goals of the current application are the development of innovative new models for basic and clinical research in mood disorders and the intensive scrutiny of 5 novel GSK antidepressant candidates in preclinical and clinical paradigms. In addition to an Administrative and Animal/Assay Core, 7 research projects are proposed. Of these, two are based in the intramural NIMH program; Neurogenesis, Synaptic Plasticity and Signal Transduction (Husseni Manji, M.D., PI) and Clinical Models to Assess Novel Antidepressants (Dennis S. Charney, M.D., PI). The remainder are based at Emory University led by established investigators including Jay M. Weiss, Ph.D. (Animal Models of Depression), Michael Davis, Ph.D. (Animal Models of Fear and Anxiety), Clinton D. Kilts, Ph.D. and Mark Goodman, Ph.D., (Functional Brain Imaging, with a focus on new PET ligand development), Michael J. Owens, Ph.D. and Charles B. Nemeroff, M.D., Ph.D. (Ex Vivo Assessment of Neurotransmitter Receptor and Transporter Occupancy of Antidepressants), and Andrew H. Miller, MD (Cytokine Induced Depression: A Rhesus Monkey Model). In conjunction with GSK, 5 novel GSK antidepressant candidates and others that become available via GSK Drug Discovery within the lifetime of the grant will be intensively scrutinized to synergize with the in-house GSK effort. This proposal encompasses virtually all of the major goals outlined in the RFA, namely, development of new neurochemical tools including novel PET ligands, exploration of new models of drug development and facilitation of a partnership between academia, NIMH and industry.

Early adverse experiences, such as childhood abuse, neglect and parental loss, have been firmly established as risk factors for the development of depression, certain anxiety disorders, and perhaps other disorders, in adulthood. We demonstrated during the previous funding period that women with a history of childhood sexual or physical abuse exhibit markedly increased hypothalamic-pituitary-adrenal (HPA) axis and autonomic responses to psychosocial laboratory stress and this effect was most pronounced in those abused women who suffered from current depression. We subsequently demonstrated alterations at multiple levels of the HPA axis, including the CNS, which together likely contribute to the increased sensitivity to stress and the manifestation of psychopathology, particularly in response to additional stress exposure. The objective of this proposed project is to confirm and extend these findings with a focus on the further characterization of the neurobiological consequences of ELS, identification of factors that might moderate or mediate increased vulnerability to stress after ELS in humans, and to expand our sample to include male subjects. We will determine whether age at the time of the ELS, and characteristics of the ELS influence responsiveness to a standardized psychosocial laboratory stressor and to a standard neuroendocrine challenge test in individuals with and without MDD. We will further determine whether the interaction of ELS and adolescent/adulthood trauma stress influences neuroendocrine reactivity in these subjects. We will also determine whether there are sex differences in the effects of ELS on neuroendocrine reactivity and whether the effects of ELS are moderated by polymorphisms of genes that are known to influence the manifestation of psychopathology in relation to life stress. The specificity of our previous observations in patients with depression will also be expanded by assessing neuroendocrine function in patients with social anxiety disorder (SAD) with and without a history of ELS, and by comparing these results to those of a new cohort of patients with major depression with and without ELS. Finally, because cognitive dysfunction in depression remains a largely unexplored research area, we will seek to determine the impact of ELS in patients with and without depression or SAD on cognitive function, i.e. perception, attention, memory, and problem solving, and whether the severity of cognitive impairment is related to HPA axis reactivity, as well as the timing and type of ELS, adulthood trauma, gender and psychopathology. This project will also serve as a human subjects core for the identification and description of individuals that will undergo assessments in Projects 0009 (Miller,PI), 0006 (Davis, PI), 0012 (Mayberg, PI) and 0013 (Kilts, PI).

psychopathology; fluoxetine; psychological stressor; neuropsychology; early experience; major depression; clinical research; human subject;

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. The objective of this study is to evaluate the effects of corticotropin releasing factor 1 receptor (CRF 1) antagonists in our measures of fear and anxiety in rats and compare it to our measure of conditioned fear. We found that the GSK CRF 1 antagonist blocks the increase in acoustic startle amplitude when rats are given CRF intraventricular or when they are exposed to bright light for long periods of time but not when startle is increased in the presence of a cue previously paired with footshock. Because the first two measures of anxiety are dependent on a brain area called the bed nucleus of the stria terminalis, but the last measure is dependent on the amygdala, we believe that the CRF antagonist may work by blocking receptors in the bed nucleus of the stria terminalis. We found that over-expression of CRF in the central nucleus of the amygdala using viral vector gene transfer leads to a persistent increase in several measures that suggests a persistent increase in anxiety.

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. The objective of this study is to develop a way to measure inhibition of fear in rhesus monkeys and humans. In rats we have developed such a procedure in which one pair of cues (AX+) is paired with an aversive airblast and another pair of cues (BX-) is not paired with an aversive airblast. Under these conditions, rats learn to be afraid of A, not to be afraid of B and fear of X is some where in the middle. Importantly, when we test fear to A in the presence of B we see that B inhibits fear to A. We have set up the AX+, BX- discrimination in humans and are testing it in patients with post-traumatic stress disorder. We found that patients with PTSD do not show normal conditioned inhibition which means we have an objective measure of one of the core symptoms of PTSD, namely the inability to feel safe. In addition, we set up the AX+, BX- in rhesus monkeys at Yerkes and are now testing it in monkeys who have had separation from their mothers for brief periods of time during development as well as in monkeys that have had lesions of various brain areas. We found that lesions of the amygdala in rhesus monkeys block acquisition but expression of fear-potentiated startle, in contrast to rats, that should a blockade of both.

[unreadable] DESCRIPTION (provided by applicant): There is considerable evidence that neurotensin (NT)-containing neuronal systems are altered in schizophrenia and mediate some of the effects of antipsychotic drugs (APDs). Much of the available literature on the manifold interactions of NT and dopamine (DA)-containing neural circuits, as well as the role of this neuropeptidergic system in the biology of schizophrenia and in the action of APDs was generated by research supported by this grant now completing its 28th year of funding. Whether NT neuronal systems play a seminal role in mediating the effects of all APDs or whether, as our preliminary data generated during the present funding period suggests, that some atypical APDs are NTergic-dependent and others are not, remains an important unanswered question. This research area is one major focus of the current proposal. Using two well characterized and validated sensorimotor gating paradigms that readily quantify APD effects, prepulse inhibition (PPI) of the acoustic startle response and latent inhibition (Ll), we determine whether APDs can be classified as NT-dependent and NT-independent. NT-dependence may represent a novel method of classifying APDs. In addition, we address the critical and related questions of which NT receptor subtype mediates the APD-like effects of NT and what characteristic of APDs renders them dependent on NT neurotransmission. Because of the lack of truly receptor specific NT receptor antagonists, the NT receptor subtype mediating the APD-like effects of NT will be identified using two innovative approaches. First, experiments utilizing viral vectors over expressing the NTT and NT2 receptors will examine the brain region specific effects of modulating NT receptor subtype specific expression. Viral vectors expressing the antisense RNA to the NTT and NT2 receptors will also be generated in order to produce brain region specific knockdown of the NT receptor subtypes. Second, the efficacy of APDs in the PPI paradigm will be compared in NT-, and NT2 receptor knockout mice. Lastly, APD effects on the NT system and APD modulation of PPI are dependent on estrous cycle stage in female rats. Therefore, the role of NT in estrous cycle regulation of PPI and APD effects on PPI will be assessed. Elucidation of the role of NTergic systems in the mechanism of action of APDs may be a critical key in understanding effective APD therapy and has the potential to assist in the development of more effective treatments of psychotic disorders. This research is particularly timely as a landmark antipsychotic drug effictiveness study (CATIE, see within) has recently been published documenting the need for novel APDs with greater efficacy and tolerability. Drugs that act upon NT circuits represent just such a novel research direction. [unreadable] [unreadable] [unreadable]

The Operations, Clinical Assessment and Administrative Core oversees this entire project and[unreadable] is responsible for the conduct of the clinical trial and the collection of biological and behavioral[unreadable] data. Based on studies of groups of major depressive disorder (MDD) patients, any single[unreadable] psychotherapeutic or pharmacological strategy in clinical practice today achieves remission in only[unreadable] a minority of patients. Thus the initial treatment an individual with MDD receives is largely an[unreadable] accident of chance rather than an evidence based choice. Over a 5-year period, 400 treatment[unreadable] naive patients with MDD will be recruited into a 12 week treatment trial of randomly assigned[unreadable] treatment with an SSRI, escitalopram, a dual 5-HT/NE re-uptake inhibitor, duloxetine or cognitivebehavior[unreadable] therapy (CBT). Subjects will be evaluated using state-of-the-art clinical assessments prior[unreadable] to, during and at the end of acute treatment period. This core describes the efforts that will result in[unreadable] the identification, recruitment of the patients, and assessment and delivery of the treatments. A[unreadable] unique component of this proposal is the study of individuals with MDD not contaminated by[unreadable] previous treatments that may confound measures of treatment response. Commensurate with the[unreadable] CIDAR goals, the data generated in this core will provide the critical foundation for hypothesis[unreadable] generation and testing in the other cores and projects.

The effort to recruit and collect multidisciplinary data on a large number of subjects is of[unreadable] most benefit when attempting to use this information to provide practical guidelines for[unreadable] clinicians treating patients. In order to accomplish this, we will need to test the possible[unreadable] interactions among separate measures such as brain function, stress reactivity and underlying[unreadable] genetic structure. The purpose of this core will be to use a combination of model building[unreadable] techniques, similar to those used in models of medical risk, and a comprehensive collection of[unreadable] relevant biological and clinical measurements to begin the process of developing reliable[unreadable] models of treatment response for depressed patients. We will use state-of-the-art variable[unreadable] selection techniques to build and validate a clinical model for overall and treatment-specific[unreadable] response, and evaluate its predictive ability through the use of statistical techniques. In[unreadable] addition, we will be incorporating genetic information, which will require specialized estimation[unreadable] techniques in order to model the relationship with treatment response. Thus we will have a[unreadable] statistical geneticist (Michael Epstein, PhD) working closely with the core to provide expertise[unreadable] and supervision for any treatment response related analyses, and a specific member of the[unreadable] core designated for handling those techniques.

[unreadable] Major depression (MDD) is a highly prevalent disease associated with significant morbidity and mortality and[unreadable] estimated to be one of the leading causes of disability worldwide. A variety of antidepressant drugs,[unreadable] psychotherapies, and non-drug somatic therapies have demonstrated efficacy in acute treatment trials.[unreadable] However, the majority of patients in these trials do not attain remission, increasing the risk for the[unreadable] development of chronic depression, suicide, substance abuse and several serious medical disorders. A[unreadable] major unmet need in the field is the identification of predictors of response to individual treatment modalities,[unreadable] as has been utilized other branches of medicine such as oncology and infectious disease, to improve patient[unreadable] outcome. In view of advances in functional brain imaging, molecular neurobiology and genetics, and a[unreadable] number of promising findings in small studies, it is propitious to conduct both hypothesis-generating and[unreadable] hypothesis-testing studies to determine whether a concatenation of factors, taken together, predict[unreadable] antidepressant treatment response. To achieve that goal, we propose a 3-arm, 12-week treatment trial led[unreadable] by Philip T. Ninan, M.D. of 400 treament naive adult depressed patients randomized to one of the following[unreadable] treatments after a 1 week placebo treatment period: 1) escitalopram, an SSRI; 2) duloxetine, an SNRI and[unreadable] 3) CBT. A series of behavioral and biological measures will be obtained that include functional brain imaging[unreadable] (fMRI) led by Helen S. Mayberg, M.D., several genetic polymorphisms led by Joseph F. Cubells, M.D., Ph.D.[unreadable] and Elisabeth Binder, M.D., Ph.D., indices of HPA axis activity, markers of immune and inflammatory[unreadable] function, as well as measures of personality, early life trauma, depression and anxiety, and cognitive[unreadable] function. In addition, in a study led by Michael J. Owens, Ph.D. both PET and an ex vivo method will be[unreadable] utilized to determine the relationship of the magnitude of SERT occupancy to clinical response in the[unreadable] escitalopram and duloxetine treatment groups and, moreover, the ex vivo method will be used to assess the[unreadable] importance of NET occupancy in treatment response to duloxetine. A Special Scientific Procedures,[unreadable] Statistical Modeling Core, led by Mary Kelley, Ph.D. will seek to determine which measure and/or[unreadable] combination of measures leads to predictors of response to the three treatments under study. Delineation of[unreadable] patient characteristics that predict treatment response to a specific treatment modality will dramatically[unreadable] improve patient outcomes and reduce the risk of inadequate treatment.[unreadable]

Although there are several effective treatments for a major depressive episode, there are no[unreadable] reliable predictors of the likelihood of remission, response or non-response with an initial trial of[unreadable] either an antidepressant medication or psychotherapy. In prioritizing a role for direct measures of[unreadable] brain functioning in the development of new algorithms for clinical management of depressed[unreadable] patients, a systematic characterization of pretreatment patterns predictive of unambiguous[unreadable] remission to standard treatments is a necessary first step. This project will characterize imagingbased[unreadable] brain subtypes that distinguish groups of never-treated depressed patients who[unreadable] subsequently respond to pharmacotherapy or cognitive behavior therapy (CBT), respectively. A[unreadable] prospectively-treated cohort of 400 never-treated depressed patients randomized to receive either[unreadable] escitalopram, duloxetine or CBT for 1.2 weeks will define these subtypes. Resting-state BOLD[unreadable] functional magnetic resonance imaging (fMRI) scans will be acquired prior to initiating[unreadable] antidepressant therapy and at a fixed, early time point specific for each treatment. Pre-treatment[unreadable] scan patterns derived using multivariate analyses and associated with the six possible response[unreadable] outcomes (3 types of response; 3 types of nonresponse) will be used to determine whether[unreadable] pretreatment brain patterns can distinguish among outcome groups. A second fMRI scan,[unreadable] acquired early in the treatment course, will be used to assess the likelihood of response to the[unreadable] specific treatment assigned. The proposed studies are a first step towards defining brain-based[unreadable] subtypes predictive of differential treatment outcome in major depression. The data from these[unreadable] studies will also be entered into more complex algorithms integrating imaging findings with[unreadable] behavioral, environmental, biochemical and genetic information for individual patients.

The overarching goal of this project is to identify hypothalamic-pituitary-adrenal (HPA)-axis-related[unreadable] parameters as potential predictors and/or biomarkers of disease progression and response to treatment for[unreadable] major depression in treatment-naTve patients. We will develop candidate parameters related to the HPA-axis,[unreadable] and more specifically, to glucocorticoid receptor (GR) function. Those parameters may include genotypes or[unreadable] haplotypes at GR-related loci, differences in expression of GR chaperone genes, measurements of GR[unreadable] function in vivo and in vitro, or some combination of these. Once our genetic investigations have identified[unreadable] putative predictors of treatment response, they will be integrated with data from neuro-imaging, transporteroccupancy[unreadable] studies, clinical assessments and other data for inclusion in an overall model of response[unreadable] predictors to be developed in the Special Scientific Procedures Core.[unreadable] The specific aims of this project include examination of relationships among HPA-axis-associated[unreadable] markers, measured at the genotypic, mRNA-expression, biochemical and systemic levels. More specifically[unreadable] we will investigate how sequence variation at the genetic level in GR receptor- regulating genes associate[unreadable] with mRNA expression in monocytes isolated from patients at multiple timepoints, and to glucocorticoid[unreadable] receptor function measured in vitro (i.e., in monocytes) as well as in vivo (using the combined[unreadable] dexamethasone suppression/CRH stimulation (DEX-CRH) test).[unreadable] Integrating multiple levels of analysis may help to identify genetic and molecular mechanisms for HPAaxis[unreadable] dysregulation and its normalization in response to anti-depressant treatment, thereby suggesting[unreadable] specific predictors for response to antidepressant treatments or disease progression. The elucidation of[unreadable] molecular mechanisms for the normalization of HPA-axis hyperactivity that accompanies successful[unreadable] antidepressant treatment may also be an important step in the development of novel antidepressants.[unreadable] This project will interact closely with the Operations and Clinical Assessment Core by coordinating all[unreadable] necessary blood draws and endocrine challenge tests and through a shared database integrating genetic[unreadable] and phenotypic data, the Research Methods Core by genotyping polymorphisms in all candidate genes[unreadable] relevant for this project and providing detailed information on their population-specific haplotypic structure,[unreadable] and the Special Scientific Procedures Core by generating multi-level HPA-axis related data for inclusion in[unreadable] the overall predictive model for treatment outcome.

Until now, determining whether patients were receiving an adequate dose of a given antidepressant[unreadable] medication was based almost exclusively upon the experience of the prescribing physician. Recently in[unreadable] vivo imaging technology with PET (positron emission tomography) and SPECT (single photon emission[unreadable] computed tomography) are emerging as new tools for assessment and optimization of pharmacological[unreadable] treatment (e.g. monitor adequacy of dosing), psychiatric medication development, and basic[unreadable] understanding of the pathophysiology of psychiatric illness. However, these techniques are associated[unreadable] with limited availability and significant financial costs that preclude the availability of this technology to the[unreadable] vast majority of clinicians. The goal of this project is to provide a valid and simpler alternative (an assay[unreadable] using a human blood sample) to PET imaging to furnish similar in vivo molecular site occupancy data.[unreadable] Briefly, it now appears that most SSRI antidepressants block approximately 80% of their target serotonin transporters[unreadable] (SERT) at standard clinical doses. This data suggests that appropriate clinical dosing might best be[unreadable] determined by assessing brain SERT occupancy. We have developed a unique method in which we are[unreadable] able to measure the magnitude of 5-HT, NE or dopamine (DA) transporter occupancy in antidepressanttreated[unreadable] patients by exposing cells transfected with the human SERT, NET or DAT to the patients' serum[unreadable] after steady-state is attained. Following validation of this technique using concomitant PET imaging we will determine what magnitude of serotonin and/or norepinephrine uptake blockade is required for an optimal treatment response.[unreadable] Simply stated, if a patient has not responded to a standard dose of an SSRI or SNRI, is it because they[unreadable] have not yet achieved a substantial occupancy of the SERT and/or norepinephrine (NET) transporter?[unreadable] These data may be extremely valuable in monitoring patient compliance, the need for dosage adjustment[unreadable] and, in the case of adequate occupancy without therapeutic response, information that provides a[unreadable] rational decision to switch medication class or initiate other treatment options.

DESCRIPTION (provided by applicant): Post-traumatic Stress Disorder (PTSD) is one of the most highly prevalent psychiatric disorders and its prevalence is likely increasing in the United States and worldwide due to the rising numbers of natural disasters (earthquakes, hurricanes, tsunamis), man-made disasters (oil spills), terrorism and wars, as well as violent crime and automobile accidents. Although the majority of trauma victims experience the cardinal symptoms of re-experiencing, avoidance and hyperarousal, for the large majority of such individuals, these symptoms do not become chronic nor do they develop syndromal PTSD. It is important to identify the large minority of trauma victims with a high likelihood of developing PTSD because of the very significant medical and psychiatric morbidity and mortality associated with this disorder. There is already considerable evidence that the likelihood of developing PTSD after trauma exposure is due to a combination of genetic and environmental factors. This two-site, linked R-01 application seeks to utilize state-of-the art advances in genomics, transcriptomics and epigenetics, coupled with comprehensive clinical and psychological measures, to address this seminal unanswered question in PTSD clinical service and research. To achieve this goal, 500 trauma-exposed subjects will be recruited at the University of Miami Ryder Trauma Center and the Emory University affiliated Grady Memorial Hospital and followed at regular intervals for one year. This focused, hypothesis-driven study will scrutinize previously identified psychological and biological risk factors. Genetic risk factors include polymorphisms of the ADCYAP1R1, FKBP5, DAT, BDNF, COMT, CRFR1, 5HTTLPR, RGS2, GABA2 and 5HT3R genes, novel genetic and epigenetic risk factors and most importantly, the primary downstream effects of these genomic and epigenetic findings by the use of conventional and newer statistical modeling methods. These findings should provide the means to identify trauma survivors who will likely develop PTSD and can therefore be referred for appropriate psychotherapeutic and/or psychopharmacologic treatment. Such a strategy has the potential to help redefine psychobiological subtypes of PTSD as well as to reduce the burden of chronic PTSD on our healthcare system.

Abstract Alcohol use disorder (AUD) with comorbid major depression (MD) is among the most frequent and serious conditions encountered in clinical practice, for which effective treatment interventions are currently limited. Activation of pro-inflammatory cytokines and pathways are emerging as key pathophysiological factors in the etiology of both alcoholism and major depression. Although immune and inflammatory mechanisms have been studied to a considerable extent in alcoholism and major depression separately, patients with co-occurring alcoholism and major depression are most likely to exhibit substantial inflammation. Pro-inflammatory mechanisms appear to play a bidirectional role in both depression and alcoholism and may underlie the limited treatment response reported in clinical trials. The lack of previously effective treatments demands new intervention modalities and different treatment targets targeting mechanisms underlying such comorbidity. We hypothesize that increased inflammation in these patients represents a major factor fostering decreased treatment response. Thus, patients with comorbid AUD and MD represent an ideal group to test the effect of a potent anti-inflammatory intervention. We propose to use allogeneic human mesenchymal stem cell (ahMSC) therapy, a highly novel treatment pioneered at the University of Miami Miller School of Medicine, to treat a variety of inflammatory conditions. This treatment has been shown to be safe and well-tolerated in a variety of medical disorders and exerts a robust and sustained anti-inflammatory effect. We plan to test the effects of ahMSCs on inflammation and on alcohol and depression outcomes in patients with comorbid AUD and recurrent MD (AUD- MD) preselected for the presence of high inflammatory markers (hsCRP>3 mg/L). Our study has the following Specific Aims: Aim 1. To examine the effects of a single ahMSC infusion on inflammation, as assessed by C- Reactive Protein (CRP) concentrations in a 12-week randomized, placebo-controlled trial in 80 MD-AUD patients (40 active infusion, 40 placebo) all preselected for the presence of inflammation. Aim 2. To examine the effects of the reduction in CRP and other inflammatory markers associated with ahMSC therapy on clinician- administered measures of the severity of alcohol use (TLFB-% heavy drinking days) and depression (MADRS) and global clinical functioning (CGI). Aim 3. To examine the direct (reduction in CRP and other inflammatory biomarkers) and indirect (reduction in alcohol use and depression) effects of ahMSC therapy on secondary study outcomes including, craving, cognition, everyday functioning and perceived quality of life. Aim 4. Our exploratory aim includes assessment of the mediating role of childhood trauma and assessment of the persistence of treatment effects over the one year follow-up period, with collection of blood samples to explore the role of inflammation related-polymorphisms and epigenetics in treatment response. If successful, this treatment would represent a paradigm shift in treating AUD-MD comorbidity and will have a significant impact on the field of therapeutics for other complex conditions in which altered inflammatory mechanisms play a substantial role in their pathogenesis.

Summary Post-traumatic Stress Disorder (PTSD) is among the most prevalent of psychiatric disorders. Trauma exposure is common, including natural disasters, terrorism, wars, automobile crashes, and violent crime. Although the majority of trauma victims experience the symptoms of re-experiencing, avoidance and hyperarousal, for the large majority of such individuals, these symptoms do not become chronic nor do they develop syndromal PTSD. It is critical to identify the underlying neurobiology of PTSD because of the very significant medical and psychiatric morbidity and mortality, and the promise of new therapeutics based on its biological underpinnings. Despite its clinical importance, there have yet to be human biology- focused postmortem studies of well-matched cases and controls to leverage the fact that this is arguably among the best understood Psychiatric Disorder in terms of neural circuit regulation. This proposal utilizes a Linked R01 mechanism across 3-sites (University of Miami (1), Lieber Institute for Brain Development (2), and McLean Hospital with Emory University (3)), to perform a postmortem, multi-omic study of brains from 300 total subjects: PTSD (civilian + military trauma, n=100), mood disorder non-PTSD psychiatric controls (n=100), and normal controls (n=100). We will focus on targeted brain regions with known differential association with PTSD risk as a function of identified intermediate phenotypes, including amygdala, prefrontal cortex and hippocampal dentate gyrus. We will determine differential DNA genotyping / methylation, RNA expression, and Proteomic patterns across brain areas. We hypothesize 1) that known pathways we have previously identified in the periphery and in PTSD models will be differentially identified in the brain regions of PTSD subjects, and 2) that genome- wide exploratory approaches will identify novel epigenetically gene pathways. Our main outcome will be a predictive model of genetic, epigenetic, transcriptomic, and proteomic profiles of brain regions from cases vs. controls. These data will allow an understanding of the region-specific genotype-dependent transcriptional and translational profiles, and findings will be integrated with detailed multi-omic data from other studies. We will use state-of-the-art statistical modeling, combined with rich biological and psychological phenotype measurements to determine a predictive model across conserved brain regions and molecular pathways. This novel, integrated, and impactful linked R01 proposal will lead to the identification of so far unknown trauma-associated genes and proteins, noncoding RNAs, and epigenetic marks in trauma related disorders and will allow the identification of novel therapeutic targets on the level of regulatory RNAs and proteins. Such a strategy has the potential to help redefine psychobiological subtypes of PTSD as well as to reduce the burden of chronic PTSD on our healthcare system.