Carol A. Shively - US grants

While coronary heart disease (CHD) is the leading cause of death in women beyond age 60, research in the area is lacking, especially concerning psychosocial risk factors, the relationship between psychosocial and other risk factors, and the efficacy of psychosocial risk factor modification. The major objective of the proposed research is to determine whether and to what extent psychosocial risk factor modification is effective in reducing the risk of diet-induced atherosclerosis in female monkeys. Other specific aims include the characterization of the nature of the relationship between atherosclerosis and social status, a psychosocial variable previously observed to be associated with atherosclerosis; and the delineation of relationships between social status and other risk factors. Data produced by this experiment will increase basic knowledge of psychosocial risk factors of CHD in females, and may have a direct bearing on the design and implementation of clinical trials of psychosocial risk factor modification. The proposed experiment requires 48 adult female cynomolgus macaques (Macaca fascicularis), fed an atherogenic diet and living in social groups of four animals each. Experimental manipulation will result in four groups: 1) females that are socially dominant before and after the manipulation, 2) females that are dominant before and subordinate after manipulation, 3) females that are subordinate before and dominant after manipulation, and 4) females that are subordinate before and after manipulation. Comparison of atherosclerosis and related behavioral and clinicopathologic risk variables among groups will provide data on the relative efficacy of psychosocial risk factor modification.

APPLICANT'S ABSTRACT: While the deleterious effects of excessive alcohol consumption are well recognized, the effects of moderate alcohol consumption are not well understood. Currently, it is thought that moderate alcohol consumption reduces the risk of coronary heart disease (CHD), increases bone density, is anxiolytic, and promotes social interaction. Thus, there currently appears to be some benefit to moderate alcohol consumption. More worrisome is the possibility that moderate alcohol consumption may increase breast cancer risk, and that it may also lead to excessive alcohol consumption. Hence, there may be a risk/benefit trade-off to moderate alcohol consumption. We propose to assess the effects of moderate alcohol consumption on CHD, osteoporosis, and breast cancer risk in ovariectomized female cynomolgus monkeys (Macaca fascicularis), a useful animal model for each of these endpoints. This animal model will allow randomized assignment to moderate alcohol or placebo treatment groups, thus avoiding the problems of self-selected populations and self-reported alcohol consumption inherent in human studies. The proposed experiment also offers complete control over alcohol dose, diet, and other variables known to affect disease endpoints. In the proposed experiment, hypothesized mechanisms of alcohol action will be addressed during the early and middle stages of disease development which is not possible using human subjects. Consequently, the use of this animal model obviates the need to wait for relatively rare clinical events to occur in order to obtain reliable measures of disease risk. Our long-term goal is to determine the relative risks/benefits of moderate alcohol consumption by simultaneous assessment of CHD, osteoporosis, and breast cancer risk.

There is increasing concern about the possible deleterious effects of menopause and potential beneficial effects of hormone replacement on cognitive function. Traditional hormone replacement therapy effectively reduces symptoms of menopause such as hot flashes, and has beneficial effects on the skeletal and cardiovascular systems, and probably the nervous system. However, hormone replacement therapy increases the likelihood of endometrial and perhaps breast cancer. The medical community is at a crossroads in the treatment of menopause. As alternatives to traditional hormone replacement therapy, novel estrogens and other treatments are being considered to identify a therapeutic intervention that has only neutral or beneficial effects on the target tissues. For this reason, it is important to compare the impact of these treatments on the function of all the target tissues of concern. The efficacy and safety of several of these alternative therapeutic strategies are currently being tested in the monkey model. Unfortunately, the monkey model is deficient in the area of assessment of cognitive function. This deficit is due to the fact that standard tests of nonhuman primate cognitive function have experimental requirements that severely constrain their use under many experimental conditions. These problems are currently addressed by evaluating cognitive performance in small groups of animals separate from those in which the effects of treatment on other tissues are being evaluated. This approach results in greater research expense and wastes valuable animal resources. Therefore, we propose to develop a test of cognitive function that can be administered on a large scale in a short time period and does not require special housing or extensive dietary manipulation. In this application, we propose to develop a short and easily administered assessment of cognitive function, determine its validity against standard tests of cognitive function, determine its reliability using a test-retest paradigm, experimentally manipulate performance on this test with a pharmacological agent known to alter cognitive function, and finally, demonstrate the utility of the test in an experimental situation in which standard assessments of cognitive function are not possible. This research will further the development of the nonhuman primate model of menopause and provide a clinical evaluation of cognitive functioning that will have wide applicability in research with nonhuman primates.

stress; dopamine receptor; social dominance; brain imaging /visualization /scanning; depression; hypothalamic pituitary adrenal axis; socioenvironment; psychological stressor; longitudinal animal study; adrenocorticotropic hormone; biomarker; social model; neurochemistry; neuropharmacology; neurophysiology; corticotropin releasing factor; behavior prediction; bioimaging /biomedical imaging; behavioral /social science research tag; magnetic resonance imaging; female; blood chemistry; positron emission tomography; Macaca fascicularis;

Abstract Not Provided.

[unreadable] DESCRIPTION (provided by applicant): CHD is the leading cause of morbidity and mortality of women in the US, exceeding that of all cancers combined. The extent of coronary artery atherosclerosis (CAA) at menopause is an important determinant of postmenopausal CHD risk. A major challenge is to identify premenopausal attributes which influence CAA extent at the menopause. This application is focused on determining the relative contribution of depression to premenopausal CAA, and whether it is a source of "reversible" risk. Although the appearance of CHD is predated by depression, suggesting that depression may be an independent risk factor for CHD, CAA develops for decades before the appearance of CHD symptoms, and the temporal relationship between depression and CAA is unclear. One common underlying mechanism implicated in both the pathology of depression and CHD is the serotonergic system. Importantly, there is initial evidence that selective serotonin reuptake inhibitors (SSRIs), commonly used to treat depression, have beneficial effects on processes that promotes atherogenesis (platelet activation, inflammation, low heart rate variability). Using premenopausal cynomolgus monkeys (Macaca fascicularis), an established model for the study of both coronary artery atherogenesis and depressive behavior, we propose to determine for the first time the temporal relationship between depression and subclinical CAA progression. Furthermore, we will examine the cardiovascular and behavioral effects of manipulating neural serotonergic function with SSRI's. Monkeys will be fed an atherogenic diet and depressive behavior recorded during an 18-month pretreatment period. Atherosclerosis extent will be determined by iliac biopsy. The monkeys then will be treated with an SSRI or placebo, balanced on pretreatment depressive behavior, and atherosclerosis progression will be assessed after 18 months. The specific aims are to determine: the magnitude of the relationship between depressive behavior and atherosclerosis; whether atherosclerosis progression is slowed by treatment targeting the neural serotonin system, or by reductions in depressive behavior, or both; pathophysiologic characteristics (e.g. platelet activation, inflammatory processes, heart rate variability, ovarian dysfunction, plasma lipids) associated with both early atherogenesis and depressive behavior; and the effects of treatment on these characteristics. The results will be relevant to public health as they will determine the relative importance of premenopausal depression to later risk for CHD and whether that risk is reversible, shape the timing and aggressiveness of antidepressant therapy, and provide evidence supporting the need for a prospective clinical trial powered to detect changes in cardiovascular outcomes with SSRI treatment. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): CHD is the leading cause of morbidity and mortality of women in the US, exceeding that of all cancers combined. The extent of coronary artery atherosclerosis (CAA) at menopause is an important determinant of postmenopausal CHD risk. A major challenge is to identify premenopausal attributes which influence CAA extent at the menopause. This application is focused on determining the relative contribution of depression to premenopausal CAA, and whether it is a source of "reversible" risk. Although the appearance of CHD is predated by depression, suggesting that depression may be an independent risk factor for CHD, CAA develops for decades before the appearance of CHD symptoms, and the temporal relationship between depression and CAA is unclear. One common underlying mechanism implicated in both the pathology of depression and CHD is the serotonergic system. Importantly, there is initial evidence that selective serotonin reuptake inhibitors (SSRIs), commonly used to treat depression, have beneficial effects on processes that promotes atherogenesis (platelet activation, inflammation, low heart rate variability). Using premenopausal cynomolgus monkeys (Macaca fascicularis), an established model for the study of both coronary artery atherogenesis and depressive behavior, we propose to determine for the first time the temporal relationship between depression and subclinical CAA progression. Furthermore, we will examine the cardiovascular and behavioral effects of manipulating neural serotonergic function with SSRI's. Monkeys will be fed an atherogenic diet and depressive behavior recorded during an 18-month pretreatment period. Atherosclerosis extent will be determined by iliac biopsy. The monkeys then will be treated with an SSRI or placebo, balanced on pretreatment depressive behavior, and atherosclerosis progression will be assessed after 18 months. The specific aims are to determine: the magnitude of the relationship between depressive behavior and atherosclerosis;whether atherosclerosis progression is slowed by treatment targeting the neural serotonin system, or by reductions in depressive behavior, or both;pathophysiologic characteristics (e.g. platelet activation, inflammatory processes, heart rate variability, ovarian dysfunction, plasma lipids) associated with both early atherogenesis and depressive behavior;and the effects of treatment on these characteristics. The results will be relevant to public health as they will determine the relative importance of premenopausal depression to later risk for CHD and whether that risk is reversible, shape the timing and aggressiveness of antidepressant therapy, and provide evidence supporting the need for a prospective clinical trial powered to detect changes in cardiovascular outcomes with SSRI treatment.

DESCRIPTION (provided by applicant): Clinical and experimental studies suggest that hippocampal volumes may be smaller in individuals with depression, although the cellular mechanisms underlying this relationship are unclear. Stressful life events are associated with an increased risk of depression, and animal models, exposed to chronic stress have been used previously to investigate hippocampal shrinkage in depression. Although the data from preclinical stress models are compelling, the degree to which stress responses in animal models are relevant to human depression remains controversial, particularly since women are at two-fold greater risk of depression and the animal models are mostly male rodents. Evaluation of the causes of reduced hippocampal volume in an experimental model that more closely resembles human depression would be valuable. We have developed a primate model of depression in adult female cynomolgus monkeys which closely resembles human depression, and recently observed that depressed monkeys have relatively small anterior hippocampi. The overall goal of this proposal is to evaluate hippocampal morphologic, cellular, and molecular characteristics in depressed and nondepressed female monkeys to determine whether the smaller hippocampi of depressed female monkeys are accompanied by reductions in neuropil and synaptic, spinous, and dendritic integrity. We have a unique and valuable collection of fixed, frozen hippocampi derived from the population of adult female monkeys in which the behavioral and physiological characteristics of depression were studied premortem for 4 years. Using the tissue from 8 depressed and 8 nondepressed monkeys we will determine astrocyte, pyramidal, and granule neuron size and number, and protein and mRNA levels of markers of synaptic, spinous, and dendritic integrity in the cornu ammonis (CA) CA1, CA2, CA3, and DG of the anterior and posterior HC of behaviorally depressed and nondepressed monkeys. The results of this study will establish the use of the model in future investigations of the mechanisms of depression and the efficacy of interventions for depression. The research is particularly responsive to the FOA entitled "Advancing Novel Science in Women's Health Research" (PAS-07-381). The results of the proposed study will be used in support of a competitive NIH application. PUBLIC HEALTH RELEVANCE: Depression is a significant health problem in the US, particularly in women, as 20% of reproductive-aged women experience clinically significant depression. Unfortunately very little research has been conducted in female animal models of depression. The use of the first primate model of adult depression in females proposed here, which has greater similarity to human neurobiology and depression than rodent models, will advance our understanding of the neurobiology of depression especially in women.

DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) is the leading cause of death and disability in women. Psychosocial stress increases CVD and CVD risk factors in women and female cynomolgus monkeys, and leads to depression and ovarian dysfunction which are especially important CVD risk factors for women. Environmental stimuli that affect CV health are initially processed by the brain which is nutrition-sensitive, modulates the function of downstream stress-sensitive systems, and may be structurally different in CVD patients. Importantly, most of the data demonstrating stress effects on CVD have been derived from subjects consuming a Western diet. In contrast, prudent diet consumption is associated with decreases in risk of depression, infertility, traditional CVD risk factors, and CVD. Emerging experimental data suggest that certain characteristics of a Western diet exacerbate physiological and behavioral stress responses, and these acute effects reflect associations linking Western diet consumption with perceived stress in population studies. Thus, psychosocial stress effects on CVD/CAA risk may not be simply main effects of stress; they may reflect diet-exacerbated stress reactivity. Here we propose to test the hypothesis that psychosocial stress-associated CVD risk is due in part to Western diet exacerbation of stress reactivity; therefore, consumption of a Prudent diet will reduce physiological stress reactivity an mitigate the deleterious effects of psychosocial stress on CVD risk. Behavioral and physiological indicators of stress and depression, food consumption and activity levels, CVD risk factors, body composition, tissue gene expression, and volumetric and other measures of key neural regions will be assessed in adult female monkeys at Baseline, and after 18 and 36 months of a Western or Prudent diet. Atherosclerosis will be measured in iliac, carotid and coronary arteries. The aims are to determine the effects of Western and Prudent diets on 1) physiological responses to an acute stressor, 2) physiological and behavioral indices of chronic stress, 3) CVD risk, atherosclerosis and artery gene expression, and 4) structural characteristics of key neural regions in socially stressed subordinate and dominant monkeys. This is a more invasive and detailed investigation of behavioral, physiological, and pathological effects of diet than can be conducted in human studies and the data from this model are more amenable to clinical translation than other animal models. If successful, the proposed studies will demonstrate that a Prudent diet can ameliorate some deleterious effects of psychosocial stress on CVD risk, and thus provide a cost-effective intervention on psychological stress with the promise of wide-spread efficacy.

? DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) is the leading cause of death and disability in women. Psychosocial stress increases CVD and CVD risk factors in women and female cynomolgus monkeys, and leads to depression and ovarian dysfunction, which are especially important CVD risk factors for women. Environmental stimuli that affect CV health are initially processed by the brain. Increasingly, the brain is understood to be nutrition- sensitive, biochemically plastic, and capable of modulating the function of downstream stress-sensitive systems. Importantly, most of the data demonstrating stress effects on CVD have been derived from subjects consuming a Western diet, a metabolic stressor which may exacerbate the effects of psychosocial stress on the system. In contrast, prudent Mediterranean-like diets have been associated with decreased risk of depression, infertility, traditional CVD risk factors, and CVD. Emerging experimental data suggest that certain characteristics of a Western (high-fat) diet exacerbate physiologic and behavioral stress responses, reflecting the associations linking Western diet consumption with perceived stress in population studies. Thus, psychosocial stress effects on CVD or atherosclerosis risk may not be simply main effects of stress; they may reflect diet-exacerbated stress reactivity. A key factor at the interface of stress, diet, and CVD risk is inflammation. Circulating monocytes are important mediators of inflammation, and are precursor cells for macrophages - central players in atherogenesis and CVD. Monocytes are sensitive to dietary and stress effects, and may provide a key nexus for understanding stress effects as well as novel targets for therapies. Here we propose to test the hypotheses that diet and stress interact to have a long term impact on cardiovascular health by epigenetically reprogramming monocyte phenotypes relevant to CVD risk, and that the consumption of a Mediterranean diet will mitigate these effects of stress. These outcomes will be evaluated in the context of transcriptomic and epigenomic phenotypes of peripheral blood monocytes and cardiovascular disease in female nonhuman primates. We will use our well-characterized female cynomolgus macaque model, which consume carefully controlled diets and are physiologically and genetically very similar to people. This study will be cost- effective because we can take advantage of the resources from an existing funded project (R01 HL 87103-06). If successful, the proposed study will provide mechanistic insights into a cost-effective intervention on psychological stress with the promise of widespread efficacy for millions of individuals at risk for CVD.

Summary This study examines the effects of diet on peripheral metabolism and central nervous system (CNS) phenotypes and pathways implicated in early-stage Alzheimer's disease (AD) pathology in nonhuman primates (NHPs). The premise for the study is based on evidence that dietary patterns are powerful modulators of brain aging. Habitual consumption of simple sugars and saturated fat, characteristic of a Western diet, is associated with chronic diseases and increased risk of AD and vascular cognitive impairment. Conversely, high intake of dietary fruits and vegetables, fish, and healthy fats, characteristic of a Mediterranean diet, is associated with reduced risk of chronic diseases, AD, and vascular cognitive impairment. Evidence supporting these associations comes from population-based studies that may be affected by confounders, or from rodent studies with limited translational relevance. Identification of specific mechanisms underlying dietary effects has been challenging to date. However, in our 1-month randomized trial in older adults comparing Western and prudent diet, we observed changes in AD and pathologic aging markers (e.g. cerebrospinal fluid A?42, lipoproteins, insulin, and oxidative stress markers), suggesting that Western diet composition may alter the brain in ways that promote AD-related neuropathologies. Nonhuman primates offer unique opportunities to model complex human diseases, and we and others have shown that they develop AD pathology and diet- related metabolic and vascular disorders with aging. The proposed study leverages a NIH-funded investigation in which adult female cynomolgus monkeys were randomized to consume a Western or Mediterranean-like diet for 30 months. Metabolic, vascular, and behavioral phenotyping, MR imaging, and serial collections of cerebrospinal fluid occurred in the Baseline and Treatment phases. At necropsy, multiple peripheral and vascular tissues were obtained and the brains collected using a protocol adapted from human AD guidelines. We now propose to use these archived tissue, images, and data to determine the effects of Western vs. Mediterranean diet on structural and functional CNS characteristics relevant to AD risk. Our overarching hypothesis is that, compared to a Western diet, consuming a Mediterranean diet protects against neuropathologic, vascular, inflammatory, oxidative, and other phenotypes associated with increased risk of AD. Our Specific Aims are to determine effects of Western and Mediterranean diets on: AD neuropathology, neurovascular pathophysiology, neuroinflammation, oxidative stress, and gene expression to identify the peripheral mediators of diet effects on the CNS, and novel pathways and mechanisms which may be involved in diet/AD interactions.

This proposal is a response to PAS-19-241, Stimulating Urology Interdisciplinary Team Opportunity Research, the aim of which is to promote innovative, high-quality, interdisciplinary research relevant to the NIDDK. Importantly, this PAS and a prior workshop identified psychosocial factors in women with urinary incontinence as an important knowledge gap. To this end, we bring together experts from behavioral sciences, urology, molecular pathology, and regenerative medicine to explore further our initial findings that socially subordinate female monkeys do not respond as well to cell therapy for urinary incontinence as their dominant counterparts. We now propose a more comprehensive approach to assess (a) sympathetic nervous system (SNS) arousal, hypothalamic-pituitary-adrenal (HPA) activation, and impaired ovarian function in socially housed monkeys; and (b) the likely pathways by which social subordination stress affects structural and functional regeneration within the urinary sphincter. Two likely pathways through which social subordination stress may modulate these processes are cortisol and SNS effects on tissue and cell damaging inflammation and estrogen-deficiency-associated inhibition of cell mobilization. These processes are not mutually exclusive and may involve the CXCL12/CXCR4 signaling pathway. Based on our previous studies and the published literature, our central hypothesis is that psychosocial stress inhibits the regenerative effects of cell therapy by reducing the mobilization of tissue healing bone marrow progenitor cells, and increasing the presence of hematopoietic tissue damaging inflammatory cells in the urinary sphincter. This hypothesis will be tested in a prospective, randomized, nonhuman primate preclinical trial using our well-characterized female cynomolgus monkey model of psychosocial stress due to social subordination, and our model of intrinsic urinary sphincter deficiency. Our Specific Aims are to determine the effect of social status on: 1) the structural (cellular, acellular, vascular, innervation) and functional (urinary sphincter and bladder) effects of autologous skMPC therapy in female primates with ISD; 2) The injected lenti-M-cherry+ skMPC cell retention in, and lenti GFP+ labeled bone marrow cell mobilization to, the urinary sphincter of dominant and subordinate monkeys; 3) The effect of social subordination stress on abundance and polarization of inflammatory cells and associated molecules in the urinary sphincter; and 4) Whether social status effects on cell therapy-induced tissue regeneration are mediated by behavioral stress, SNS, HPA, or ovarian function. The results of this translational research will promote understanding of limitations and potential future regenerative medicine strategies for women with urinary incontinence.

ASTRACT Non-human primates (NHP) such as baboon, cynomolgus macaque, rhesus macaque, and vervet are susceptible to the same age-related health challenges and diseases as humans. The overall aging trajectory in NHP is also influenced by stressors and lifestyle similar to humans. Research in NHP allows for controlled environments, providing longitudinal data with less ?noise? than in human studies. The genetic, metabolic and physiologic similarities between NHP and humans make findings in NHP directly translatable to our understanding of human disease processes, including genetic predispositions and early molecular indicators of these processes. In the R21 phase of this project, we will 1) harmonize existing samples and data in four NHP species that can be leveraged to better understand molecular and cellular mechanisms underlying human aging, and 2) implement the Monkey Inventory and Data management of Samples (MIDAS), a LabKey Server data management system, for integration of omic data and pedigree data with clinical and research data from Aim 1. In the subsequent R33 phase, we will demonstrate the unique value of this harmonized, comparative cross- species resource for aging studies using integrated omics and clinical measures to quantify aging in liver and plasma across the lifespan in these four NHP. We will 1) determine the biological age trajectory for each NHP species from multiple chronological ages that capture human equivalent 18-80 years, by measuring molecules known to reflect biological age including gene expression, DNA methylation, and proteins in liver samples (n=48 for each species (F, 24; M,24)); and 2) identify circulating molecular signatures that correlate with liver specific signatures of aging that precede clinical markers of aging common to the four NHP species Alignment and harmonization of data and samples for these four NHP cohorts will provide a critical resource translating discoveries to humans, and supporting interdisciplinary studies of aging across the lifespan.