Caleb E. Finch - US grants

DESCRIPTION: (Adapted from the application) The applicants propose a continuation of this training program in the Neurobiology and Endocrinology of Aging, which was established in 1982 by CE Finch (Director, PI) and R Bergman (Co-Director, Co-PI). Their goal is to recruit outstanding trainees who will become leaders in biomedical gerontology. They understand that integrative approaches are required for resolving the basic mechanisms in human aging, particularly in regard to the endocrine and neural systems which interface with virtually all aspects of human aging. Training is available in disciplines, technologies, and model systems that comprise the state-of-the-art in biogerontology. They offer a broad range, from mathematical modeling of hormone responses and of learning, to transgenic animals, to population genetics, to psychoneuroendocrinology. Pre- and postdoctoral trainees attend bimonthly meetings with Preceptors, which include short reviews of the laboratory program by the Preceptor, followed by a data- oriented "chalk-talk" by 2-3 trainees. Clinical presentations allow Trainees to interact with patients afflicted by Alzheimer s disease, stroke, and other neurological conditions of aging that give a foundation for developing applications to human health in the later years. Support is requested for 8 predoctoral and 8 postdoctoral fellows to be trained by 24 tenure-track faculty Preceptors who represent established predoctoral graduate programs: Neurosciences; Molecular Biology (both in the USC College of Letters, Arts, and Sciences); Physiology/Biophysics; in Pathology (both in the USC School of Medicine); and Molecular Toxicology/Pharmacology (USC School of Pharmacy). Minority recruitment was expanded in conjunction with Cal State LA, a neighboring 4 year college with a MS program.

These studies concern the neuroendocrine impairments associated with reproductive aging in C57B1/6J mice. The roles of the ovary will be analyzed in two respects: 1. the contribution of the reduced number of ovarian follicles to the early changes in reproductive cycles, and 11. the role of disturbed endocrine regulation in causing hypothalamic-pituitary impairments. Experiments will test the reversibility of age changes by long-term ovariectomy and the intensification of age changes by treatment with estradiol (E2). Neuroendocrine function will be assayed by the LH surge, the performance of ovarian grafts, hypothalamic monoamine metabolism nuclear receptors for E2 in the hypothalamus, and hypothalamic neuronal number. These studies bear on the etiology of pituitary tumors of adult humans (in which disturbances of phyothalamic monoamine metabolism are implicated), on the ovarian role in menopausal dysfunctions, and on possible but yet unidentified effects of chronic steroid usage in humans through contraceptives and postmenopausal therapy.

This program focuses on age changes in dopaminergic regulation in rodent models which are pertinent to idiopathic and drug-induced basal gangliar diseases of human aging. There are three interactive projects. (I) C.E. Finch. Characertization of dopaminergic receptor age changes by biochemical and endocrinological approaches to determine early events in agining. (II) P.K. Randall. Electrophysiological and behavioral characterization of dopaminergic receptor changes during aging. (III) D.S. Linthicum and M.B. Bolger. Development of immunologic probes (anti-idiotypic antibodies with dopaminergic specificities) which will be used to characterize dopamine receptor subtypes in projects I and II.

Alzheimer's disease;

Gene expression in brain cells during Alzheimer' disease (AD) and aging will be studied to resolve changes in relation to likely causes that include transynaptic responses, interactions of steroids with neuronal degeneration, and glial reactivity. Two projects and 3 pilot grants are proposed by the PI and junior investigators in the research group (Drs. Patrick may, David Morgan and Nancy Nichols): 1. Analysis of mRNA changes during hippocampal and cortical neuron sprouting as response to damage afferents (C.E. Finch and P. May, PI) 2. Mechanisms of glial hyperactivity (D. Morgan, PI) Pilot Projects 1. Messenger RNA and Interventions into Neuronal Atrophy (S. Johnson and C.E. Finch PI) 2. Corticoids, Glutamine Synthase and Neurodegeneration (P. May and C.E. Finch, PI) 3. Stress-related Changes in Brain RNA of Vervet Monkeys (N. Nichols and C.E. Finch, PI) Comparisons will be made between changes in postmortem human brain and rodent models, in which we will test the manipulatability of the modelled lesions. These projects will also evaluate the long- standing but inadequately tested view that there are major impairments of RNA and protein synthesis during aging and AD. In addition, the findings bear on the selectivity in neurodegeneration during aging and AD: The apparently maintained functions of some neurons gives a basis for optimism that advancing age is not a sufficient cause of neurodegeneration, and that specific mechanisms may be found to prevent or reverse the degenerative changes. Members of this program will also train pre-and postdocs in experimental approaches to age-related neurodegeneration, and will give lectures to undergraduates in the pre-health sciences major of the Leonard Davis School of Gerontology.

Partial support is requested for presentations at the 1992 annual meeting of the Gerontological Society of America (GSA), the Biological Sciences Section, Nov. 18-22, Washington DC. The grant will support travel and local expenses of speakers in symposia and special lectures during this 4 day meeting. The overall theme of the GSA meeting will be Health Challenges of an Aging Society, while that of the Biological Sciences Section will be Multi-level Mechanisms of Biological Aging. There will be 9 Symposia with 4 speakers each, which were designed to maintain a balance between established and emergent research areas. The meeting will be organized to maintain continuity with major ongoing research areas, but also will introduce new themes that have potential bearing on the emergent lines of investigation. Plenary Lectures will be given on cross-cutting themes that will benefit from critical review and new synthesis by a single scholar. The 5 Plenary lectures will be interspersed between the Symposia, Individual Slide Presentations, and the Poster Sessions. One day will be devoted to Workshops which will give a forum for short presentations and informal debates. The topics of these 2.5 hour Workshops will include: (1) In vitro cell aging; (2) Neuroendocrine and autonomic aging; (3) Comparative approaches; (4) Gene expression and Genomic stability. Following short introductory perspectives by Workshop Chairs, there will be presentations by speakers selected from the Abstracts submitted to GSA that represent different viewpoints on ongoing research issues. The State-of-the Art in Biogerontology for Non-biologists will be presented through Lectures and Symposia that present material from molecular genetics to organ function in humans and a range of animal models. Among the topics will be discussions of genetic engineering approaches to analyzing genes for Alzheimer disease and the issues about the levels of certainty in genetic identification of risk factors in Alzheimer and other diseases. Some speakers will also participate in the other events.

This ADRC consortium represents more than 100 faculty-level Participating Investigators with major interests in Alzheimer disease and the neurobiology of aging. Most studies are performed in facilities for basic and clinical research at the University of Southern California (USC) and the University of California at Irvine (UCI). Basic research emphasizes molecular mechanisms in neurodegeneration. Clinical research emphasizes new diagnostics and interventions for AD victims and their care-givers. Besides the required Cores [Administrative, Clinical, Neuropathology, and Research Training and Information Transfer Cores], we have Cores for Genetic Epidemiology, Pharmacology, and a Satellite at Cal State Los Angels (CSLA). A new Neuro-imaging Core is proposed. Program at CSLA and in other Cores address minorities that are under-represented in research on Alzheimer's disease, in order to increase access of the victims to new diagnoses and treatments, as well as to reduce the burden of care-givers. A common database is being constructed through the Teradata relational database computer at UCI that will be accessible to researchers throughout this ADRC.

Gene expression in brain cells during Alzheimer' disease (AD) and aging will be studied to resolve changes in relation to likely causes that include transynaptic responses, interactions of steroids with neuronal degeneration, and glial reactivity. Two projects and 3 pilot grants are proposed by the PI and junior investigators in the research group (Drs. Patrick may, David Morgan and Nancy Nichols): 1. Analysis of mRNA changes during hippocampal and cortical neuron sprouting as response to damage afferents (C.E. Finch and P. May, PI) 2. Mechanisms of glial hyperactivity (D. Morgan, PI) Pilot Projects 1. Messenger RNA and Interventions into Neuronal Atrophy (S. Johnson and C.E. Finch PI) 2. Corticoids, Glutamine Synthase and Neurodegeneration (P. May and C.E. Finch, PI) 3. Stress-related Changes in Brain RNA of Vervet Monkeys (N. Nichols and C.E. Finch, PI) Comparisons will be made between changes in postmortem human brain and rodent models, in which we will test the manipulatability of the modelled lesions. These projects will also evaluate the long- standing but inadequately tested view that there are major impairments of RNA and protein synthesis during aging and AD. In addition, the findings bear on the selectivity in neurodegeneration during aging and AD: The apparently maintained functions of some neurons gives a basis for optimism that advancing age is not a sufficient cause of neurodegeneration, and that specific mechanisms may be found to prevent or reverse the degenerative changes. Members of this program will also train pre-and postdocs in experimental approaches to age-related neurodegeneration, and will give lectures to undergraduates in the pre-health sciences major of the Leonard Davis School of Gerontology.

This project addresses steroidal influences of synaptic remodeling in the rodent brain in vivo and in vitro with cell culture models, with a focus on astrocyte genes that are regulated by estradiol (E2) and corticosterone. Little is known about how steroids modulate gene expression in relation to spontaneous and injury-induced synaptic remodelling. This information is crucial to optimizing estrogen replacement therapy (ERT) in the prevention and treatment of Alzheimer disease (AD). We emphasize the regulation of GFAP and other mRNAs in the hippocampus, a brain region that is well studied for synaptic plasticity in association with learning and memory and in response to lesioning, but hat also shows a physiological synaptic remodelling during the rat estrus cycle. The regulation of GFAP receives most attention, because of its sensitivity to sex steroids, glucocorticoids, and lesions, and because of prominent age-related increases in GFAP mRNA and protein. Other responses to lesions and steroids include genes expressed in astrocytes that mediate lipoprotein trafficking and genes expressed in neurons that encode proteins of the cytoskeleton and growth cones. As an efficient in vitro model for steroid-lesion interactions, we are studying "wounding-in-a-dish" with montypic cultures of primary astrocytes, with and without co-cultured neurons. In vivo studies with female rats examine astrocyte- and neuron-expressed genes in response to E2 treatment, perforant path lesions, and E- lesion interactions. Aging rats will be examined for select responses. To probe the basis for the protective effects of ERT in AD, we will use these models to compare the effects of Premarin and its equine estrogen constituents with those of E2.

Cognitive functions and brain cell activities show responses to estrogens that give a general basis for the protective effects of estrogen replacement therapy (ERT) in Alzheimer disease (AD). A team of USC investigators proposes experiments on rodent models in vivo and in vitro to analyze mechanisms by which estrogens interact with neurons and glia to modify synaptic plasticity. We will test the hypothesis that estrogens are neuroprotective by examining molecular, cellular, and physiological responses to estrogen and interactions with lesions in rats as a function of aging. We also examine bioactivities of equine estrogens that are constituents of Premarin, the most widely used ERT.

Administration-Biostatistics (Core A) assists investigators located at two USC campuses; the University Park ("main") campus and the Health Sciences campus. Core A organizes monthly data sessions among the Program Project Investigators and their trainees in three NIA-sponsored training programs. Annual meetings will be held with the External Advisory Committee. Core A interfaces the four projects with investigators of other major research communities at USC, particularly clinical studies of estrogen in Alzheimer disease and vascular dementia. The Core maintains records of expenditures and assists in preparation of annual progress reports. The Biostatistics Module (David Lavond) maintains a database for information derived from shared tissues and advises on statistical analysis for individual projects and on the data generated from shared tissues between projects.

biomedical facility; genetically modified animals; laboratory rat; laboratory mouse;

This renewal of AG-05142 proposes an Alzheimer Disease Research Center (ADRC) program seated at USC with 4 aims and 4 new projects. Aim 1: Identify molecular and cellular mechanisms of neurodegeneration during D and normal aging processes. Aim 2: Develop hormonal interventions into age-related cognitive changes with clinical studies and animal models. Aim 3: Develop cognitive & imaging techniques to monitor aging, AD, & clinical interventions. Aim 4: Increase participation of Hispanic, African-American, and other minorities in drug trials. The USC-ADRC is comprised of the required cores and two special-service cores*: Clinical Core: Dir., Victor Henderson; co-Dir. Lon Schneider (Pharmacology Program). Minority Diagnosis and Treatment Satellite*, Dr. Helena Chui. Neuroimaging Core*, Dr. Manbir Singh. Neuropathology Core, Dir. Carol Miller. Education Information Transfer Core, Dir, Bob Knight. These cores serve four new projects of USC faculty: Proj. Alzheimer-like pathology in inclusion-body myopathy (Valerie Askanas). Proj. Choroid plexus in Abeta/apolipoprotein CNS homeostasis (Berislav Zlokovic). Proj. Aberrant DNA synthesis in dividing and non-dividing cells (Myron Goodman). Proj. fMRI studies of working memory, estrogen, & aging (MaryEllen MacDonald and Manbir Singh). The Cores also support a collaborative longitudinal study with Kaiser Permanente: Alzheimer disease & estrogen replacement; Galen Buckwater, co-PI) and two NIA program projects: Models of estrogen interactions with AD and The aging brain: vasculature, ischemia, & behavior. Genotypes of human subjects are obtained through the Molecular Biology Core of the NIEHS enter (John Peters, Dir.) Other users of these resources includes >40 federally funded USC faculty (Schools of Medicine; of Pharmacy; of Gerontology of Letters, Arts, & Sciences; and of Engineering), as well as >20 non-USC researchers. Collaborations with ADRCs of UCLA, UC Irvine, UC San Diego, Boston U, Columbia, Emory, Washington U, U Washington. These approaches to AD build on AD build on USC's long-standing commitment to research and education on aging.

This renewal of AG-05142 proposes an Alzheimer Disease Research Center (ADRC) program seated at USC with 4 aims and 4 new projects. Aim 1: Identify molecular and cellular mechanisms of neurodegeneration during D and normal aging processes. Aim 2: Develop hormonal interventions into age-related cognitive changes with clinical studies and animal models. Aim 3: Develop cognitive & imaging techniques to monitor aging, AD, & clinical interventions. Aim 4: Increase participation of Hispanic, African-American, and other minorities in drug trials. The USC-ADRC is comprised of the required cores and two special-service cores*: Clinical Core: Dir., Victor Henderson; co-Dir. Lon Schneider (Pharmacology Program). Minority Diagnosis and Treatment Satellite*, Dr. Helena Chui. Neuroimaging Core*, Dr. Manbir Singh. Neuropathology Core, Dir. Carol Miller. Education Information Transfer Core, Dir, Bob Knight. These cores serve four new projects of USC faculty: Proj. Alzheimer-like pathology in inclusion-body myopathy (Valerie Askanas). Proj. Choroid plexus in Abeta/apolipoprotein CNS homeostasis (Berislav Zlokovic). Proj. Aberrant DNA synthesis in dividing and non-dividing cells (Myron Goodman). Proj. fMRI studies of working memory, estrogen, & aging (MaryEllen MacDonald and Manbir Singh). The Cores also support a collaborative longitudinal study with Kaiser Permanente: Alzheimer disease & estrogen replacement; Galen Buckwater, co-PI) and two NIA program projects: Models of estrogen interactions with AD and The aging brain: vasculature, ischemia, & behavior. Genotypes of human subjects are obtained through the Molecular Biology Core of the NIEHS enter (John Peters, Dir.) Other users of these resources includes >40 federally funded USC faculty (Schools of Medicine; of Pharmacy; of Gerontology of Letters, Arts, & Sciences; and of Engineering), as well as >20 non-USC researchers. Collaborations with ADRCs of UCLA, UC Irvine, UC San Diego, Boston U, Columbia, Emory, Washington U, U Washington. These approaches to AD build on AD build on USC's long-standing commitment to research and education on aging.

On September 22-23, 2000 (Fri-Sat), Caleb E. Finch and Robert E. Ricklefs plan to convene a Symposium on Organisms with Slow Aging (SOSA) at the Andrus Gerontology Center, University of Southern California. SOSA will critically examine the emerging evidence that some multi-cellular organisms have evolved very slow rates of aging with anti-aging mechanisms that are pertinent to human aging processes. Rates of aging can be evaluated by recognized criteria: demographic (age-specific mortality rates and fecundity); molecular (oxidative damage and telomere DNA length); and cellular-physiological changes, e.g. clonal life span, cell turnover, and organ functions. Examples from vertebrates, invertebrates, and vascular plants show a range of long life spans, which overlap with, or exceed, the upper ranges of human life spans. Field data for mortality and reproduction are available for certain in long-lived fish, turtles, and birds. The complex biology of long-life spans will be discussed in terms of evolutionary theory. Speakers will identify sources of data and availability of biological specimens to stimulate research and attract new scientists and trainees. The program includes a volunteer poster session. The specific aims of the meeting are to find a common basis for discussing the biology of long life spans in a variety of animal and plant models; organize the building of a comparative database for demographic and physiological aging parameters; and recruit new investigators and trainees to the comparative study of organisms with extended reproductive schedules and extreme longevity.

Cognitive functions and brain cell activities show responses to estrogens that give a general basis for the protective effects of estrogen replacement therapy (ERT) in Alzheimer disease (AD). A team of USC investigators proposes experiments on rodent models in vivo and in vitro to analyze mechanisms by which estrogens interact with neurons and glia to modify synaptic plasticity. We will test the hypothesis that estrogens are neuroprotective by examining molecular, cellular, and physiological responses to estrogen and interactions with lesions in rats as a function of aging. We also examine bioactivities of equine estrogens that are constituents of Premarin, the most widely used ERT.

This renewal of "Training in the Neurobiology and Endocrinology of aging" proposes to support 8 predoctoral and 8 postdoctoral trainees under the direction of 25 tenure track faculty from the School of Gerontology; the College of Letters, Arts, and Sciences; the School of Pharmacy; and the School of Medicine. Effort is given to increasing the recruitment of under-represented minorities. The training program is based on directed research in the laboratory; bimonthly chalk-talks; a new seminar course: "Recent advances in the neurobiology and endocrinology of aging"; and instruction in the responsible conduct of research. Opportunities for human studies on aging, including ethnic differences in aging processes, are provided through the Alzheimer Disease Research Center and the Diabetes Research Center.

DESCRIPTION (provided by applicant): Core A serves general administrative functions and is the seat of the Biostatistics module and a new web site. The Core distributes information (articles of interest, seminars, etc.) and organizes monthly meetings to support projects on two campuses. Most personnel and project directors are in nearby labs on the University Park Campus, except for Dr. Brinton, whose lab is on the Health Sciences Campus. Meetings with External Advisors are convened every 18 months. Investigators of this PO1 also give presentations to trainees of several Training Programs and to the USC Alzheimer Center. The Biostatistics module (Dr. David Lavond) organizes data management and archiving and maintains the web site.

Apolipoprotein J (clusterin, SGP-2) expression is increased during Alzheimer disease (AD) and to a lesser extent during normal brain again. New data shows that apoL can activate microglia in vivo and in vitro. We propose the hypothesis that apoJ expression is an adverse factor in AD and aging. In vitro, apoJ blocks the fibrillogenic pathway of AB aggregation and favors the formation of soluble toxic oligomers (ADDLs). New data show that ADDLs arise in AD brains. The effect of apoJ will be tested by infusion of apoJ into APP transgenic mice and in begenic apoJ-deficient x APP over-expressing mice. We will manipulate aging and AD-like changes in APP transgenics with caloric restriction and ibuprofen. We also predict that apoJ-deficiency will attenuate microglial activation during normal aging. In vitro studies with primary glial cultures from wildtype and apoJ-deficient mice will examine mechanisms of apoJ in microglial activation and in the sensitivity of neurons to AB.

Alzheimer's disease;

DESCRIPTION (provided by applicant): This program project grant, entitled "Models of estrogen action in Alzheimer disease," addresses age changes in brain cell responses to sex steroids that are pertinent to Alzheimer disease, with a focus on estradiol (E2). The five projects investigate components of the general pathway of E2 effects on astrocytes and neurons, using mice that lack sex steroid receptors or src. Project 1: Astrocytes, estradiol, and aging in hippocampal plasticity; C Finch and I Rozovsky Project 2: Estrogenic steroids: neurotrophic action and mechanism; R Brinton and T Berger Project 3: Sex hormones, synaptic plasticity, aging and stress; R Thompson and M Foy Project 4: Estrogen and glutamate receptors in plasticity and AD; M Baudry Project 5: Androgens and neuroprotection in AD models; C Pike Core A ( Finch) includes a Biostatistics Module and Database (D Lavond) for data obtained on the same animals by separate projects Core B (Animals, Finch) provides aging rodents of defined hormonal status (Sprague-Dawley rats; C57BL/6J mice) and genotyped mutant and knockout mice (Er-alpha-KO, Er-beta-KO, Tfm (mutant androgen receptor), and src-KO).

DESCRIPTION (provided by applicant): This revised proposal (R21 AG025496) aims to develop new mouse models to understand the neurobiology and neuropathology of C-reactive protein (CRP). Blood CRP, the major acute phase reactant detected during an inflammatory response, is a robust risk predictor for first-ever or recurrent coronary events. CRP elevation is also associated with cerebral ischaemia and may be a contributing factor in Alzheimer disease (AD) and vascular dementia. Because inflammatory processes are active in aging, AD, in vascular disease, we hypothesize that CRP elevations will accelerate glial activation during normal brain aging and will intensify AD-like changes. This hypothesis will be evaluated with transgenic mouse models which have chronic CRP elevations, either systemically or specifically in neurons. We propose four aims: Aim 1: Determine effects of chronically elevated blood human CRP (huCRP) on glial activation during normal brain aging, using existing CRP overexpressing mice (huCRPtg). Aim 2: Construct mice with neuron-specific expression of transgenic huCRP (neuron-huCRPtg) Aim 3: Study contribution of huCRP elevations to brain lesions in young neuron-huCRPtg mice. Aim 4: Study contribution of hu CRP elevations in existing AD transgenic mice (APPswe/ind) that rapidly accumulate deposits of Abeta-amyloid. F1 hybrids will be made with APPswe/ind and the existing systemic huCRPtg (Y1) and with the neuron-specific huCRPtg (when available). The F1 hybrids will be examined for changes in deposits of Abeta-amyloid.

Progesterone, Glial Aging, and Alzheimer Disease In this Program Project we jointly propose to examine the effects of progesterone (P4) and estrogen (E2) on synaptic plasticity in rodent brain. In this project, we hypothesize that progestogens directly regulate glial activities via progesterone receptor/s and via interactions with E2-dependent pathways. We will examine effects of P4, and selected progestogens including medroxyprogesterone (MPA), on glial support of neurite sprouting and repair. Pilot data show P4 supported neurite sprouting in astrocyte/neuron co-cultures. However, when microglia were added to the co-culture (mixed glia/neuron), P4 did not support sprouting and reduced neuron survival. Moreover, P4 attenuated E2-dependent sprouting in mixed glia/neuron co-cultures. These observations suggest that P4 may regulate microglial-mediated inflammatory mechanisms. Because aging also influences sprouting and inflammation, we will examine the role of glial age in P4-E2 interactions, using in vivo and in vitro models for aging and Alzheimer's Disease (AD). New in vivo models for hormone therapy (HT) will be developed including perimenopausal and postmenopausal equivalents in the rat and in the triple transgenic AD mouse. In the perimenopausal model, HT will be given to middle-aged female rats characterized by irregular cycles. The menopausal model will address one of the critical questions in human HT - is HT effectiveness dependent on the lenght of time after menopause that HT is begun? These newly developed models will provide insight into the basic mechanisms underlying the effects of HT. In vitro models will include glial-neuron co-cultures of "wounding-in-a-dish" and glial-induced neurotoxicity, using glia derived from adult rat brains of different ages.

The Animal Resources Core (Core A) will provide young and middle age Sprague-Dawley rats of defined ovarian status used by all projects. Daily vaginal cytology will identify middle-aged irregular cyclers as a perimenopausal model. Some animals will be OVX and given replacements of estradiol (E2) with or without progesterone (P4). Necropsy will identify gross organ pathology, e.g. pituitary tumors. All projects also use the Alzheimer disease (AD) triple transgenic Alzheimer mouse: females of different ages will be OVXs and given steroid replacements. Tissues will be collected and deposited into storage managed by the Analytic Core. Blood will be given to the Analytic Core to be assayed for E2 and P4. A main goal of the animal core is the development of the rodent perimenopause and menopause model. These latter animal models will be crucial in determining the impact of reproductive aging on the neural responses to hormone therapy.

Alzheimer disease (AD) involves infiammatory processes at many levels. Because ovarian steroids show many anfi-infiammatory activities, hormone therapy (HT) in AD may involve inflammatory pathways activated during the menopause transitions when hormonal deficits emerge progressively. We propose rodent models of perimenopausal stages to analyze inflammatory changes in brain regions vulnerable to AD, with a focus on aging Sprague-Dawley rats. The 3xTg-AD mouse model will be used to examine specific hypothesis about relationships of inflammation during aging to human familial AD. These models will be examined for responsiveness to HT, according to the KEEPS clinical model of E2 + cyclic P4. We will examine E2-P4 effects on microglial-astrocyte interacfions in infiammafion and in neurotrophic effects. Specific Aim 1: Infiammatory profile in rat and mouse models of human perimenopause and menopause stages (Animal Core B). 1.1. Inflammatory gene expression and bioinformafic analysis in hippocampus and cerebral cortex of aging rats and mice from different menopausal stages (Analytic Core). 1.2. In vitro modeling of perimenopausal inflammatory changes, using primary glial cultures originated from perimenopausal rats. 1.3. Bioenergefics of glia cultured from perimenopausal rats (with Project 1). Specific Aim 2: Neurodegenerative mechanisms in perimenopausal infiammatory changes, using glial- neuronal co-cultures. 2.1. Role of infiammatory genes in glial-mediated neuronal support. 2.2, Role of estrogen receptors (ERa:ER|3) in regulating perimenopausal glial inflammatory phenotypes. Astrocytes from aging rats are deficient in neurotrophic support, in which one factor is altered ratio of estrogen receptors ERa:ERp (pilot data). 2.3. Role of progesterone receptors (PR) in regulafing perimenopausal glial infiammatory changes, with a focus on Pgmrcl (pilot data). 2.4. Role of glial N0S2 in neuronal mitochondria dysfunction (with Project 1). 2.5. Obesity-infiammation interactions (with Project 3). Specific Aim 3: Effects of perimenopausal HT on brain cell infiammation. Following the KEEPS model of E2 + cyclic P4, rats and 3xTg-AD mice will be given HT during perimenopausal cycling stages. 3.1. Infiammatory gene expression and bioinformafics (Analytic Core). 3.2. In vitro models of infiammatory changes, using primary glial from aging rat cerebral cortex. 3.3. Bioenergefics of glial cultures (with Project 1). RELEVANCE (See instmctions): Our primary goal is to develop animal models for the perimenopause transition to analyze brain cell sensitivity to ovarian steroids. Findings are relevant to optimization of hormone therapy to protect against normal age-related cognitive decline, multi-infarct demenfia and Alzheimers Diease

DESCRIPTION (provided by applicant): Air pollution may accelerate brain aging. Epidemiological studies of the middle-age have shown accelerated cognitive decline in association with local differences in air pollution (Chen & Schwartz 2010), while brains of young adults from a highly polluted Mexican city had premature deposits of diffuse amyloid and glial inflammatory activation which were greater in apoE4 carriers (Block & Calderon-Garciduenas 2009). Corresponding rodent models with defined exposure to traffic-generated nano-sized particulate material (nPM) have shown glial inflammatory changes. Our pilot data show that exposure to nPM for 10 weeks activated glia and altered neuronal glutamatergic receptors in vivo and in vitro. We propose to analyze effects of nPM on glial inflammation, amyloidogenesis, and neurodegeneration in J20-ADtg mice and in triple transgenic AD mice (3xTg-AD), which has more extensive AD-like neuropathology. We will also test the hypotheses that nPM promotes Abeta42 production and exacerbates Abeta42-induced toxicity using in vitro models.

DESCRIPTION (provided by applicant): Air pollutants derived from fossil fuels are recognized for accelerating arterial and pulmonary diseases. Moreover, brain dysfunctions during development and aging have been associated with air pollutants by postmortem findings on young humans (Block & Calderon-Garciduenas 2009) and by epidemiologic studies of the middle-aged (Chen & Schwartz 2009). Correspondingly, young rodents show glial inflammatory changes with defined exposure to traffic-generated nano-sized PM (nPM) (Zanchi et al 2010; Kleinman et al 2008). Our pilot data show effects of nPM on neuronal glutamatergic receptors in vivo and in vitro. We propose to analyze age and gender interactions in neurodegenerative effects of nPM in young adult and middle-aged C57BL/6 mice, using re-aerosolized nPM from urban Los Angeles. Aim 1 will expose animals to re-aerosolized nPM for 20 weeks, with assessment of memory, and neuronal and glial functions. Aim 2 will evaluate in vitro glial age effects on neuronal activities in response to aqueous suspensions of nPM.

? DESCRIPTION (provided by applicant): Amyloid and inflammation: modulation by apoE, gender, air pollution, and drugs. We propose to test novel inflammation-gender-environment interactions on brain amyloid and microbleeds (microhemorrhages) in mouse models for Alzheimer disease (AD) and aging. The highest AD risk group, women apoE4 carriers, is modelled in EFAD mice carrying familial dominant AD genes in combination with human apoE alleles (targeted replacement, ApoE-TR). Female E4FAD mice have the most brain cytokine induction, amyloid peptide (Abeta) accumulation, and microbleeds (pilot data). In humans, cerebral microbleeds are most prevalent in apoE4 carriers, and are associated with higher conversion to clinical AD. Among environmental influence on cognitive aging and AD, urban air pollution is also considered in an experimental model with nano-scale particulate matter (nPM) from an urban traffic air corridor (with Constantinos Sioutas, co-Investigator). The nPM of well-characterized composition are re-aerosolized for controlled exposure of mice. Pilot data show EFAD mice respond with apoE allele specificity. Aim 1 addresses the age schedule of emergence of gender-apoE allele interactions in AD phenotypes during postnatal development and aging. EFAD brains will be studied for inflammatory responses (TNFa, microglia); for APP processing and Abeta deposits; for cerebrovascular amyloid and microbleeds; and for spatial memory (with Christian Pike, co-Investigator). Sex differences will be manipulated by neonatal steroid treatment to test the hypothesis that excess expression of Abeta, in either genetic sex, will drive the level of inflammation and microbleeds. In vitro, EFAD microglia will be studied for apoE-sex interactions on inflammatory gene expression. Aim 2 examines air pollution nPM effects in female EFAD mice by dose and time on accelerating brain inflammation and AD phenotypes. Critical ages of nPM exposure for AD-phenotypes will be examined with findings from Aim 1 for the earliest age showing gender differences in AD-phenotypes. We can only test one sex because nPM treatment doubles the number of variables. We prioritize females in this aim since females are the highest AD risk group. However, in vitro studies of microglia and neurons from adult brains will include both sexes for nPM induced inflammatory and pro-amyloidogenic responses. Aim 3 examines a drug intervention for brain inflammation and microbleeds by a novel gamma secretase modulator to attenuate Abeta synthesis. Drug-treated female EFAD mice will be exposed to chronic nPM and examined for apoE interactions on AD-phenotypes.

? DESCRIPTION (provided by applicant): Prior work has associated air pollution with accelerated atherosclerosis and with the incidence of stroke. Moreover, recent epidemiological studies associate air pollution with accelerated cognitive decline of normal aging. Associations with Alzheimer disease are being studied, supported by postmortem findings that humans exposed to extreme urban pollution have premature brain inflammation and deposits of diffuse amyloid-beta (Abeta). Rodent models support these emerging associations of cognitive dysfunctions with air pollution. After exposure to various urban air pollutants, several rodent genotypes show synaptic changes and increased Abeta, as well as induction of TNFa and other inflammatory responses. Our model uses chronic exposure of mice to nano-scale PM (nPM) derived from urban traffic, which shows greater cytotoxicity than larger PM. Under controlled exposure by duration and dose (nPM density), mice show a 30% decrease of the hippocampal GluA1 AMPA receptor subunit, with little change in other AMPA receptor subunits, or in NMDA receptor subunits, while in vitro studies show that nPM exposure increases excitotoxic damage. Because of its role in fast excitatory transmission and in spatial memory, GluA1 decreases could contribute to cognitive decline in association with air pollutants. Another potential link to brain aging involves interactions of AMPA receptor subunits with Abeta. This proposal addresses key gaps in experimental models for epidemiological associations and postmortem findings: the impact of nPM exposure on hippocampal neuronal physiology associated with cognitive processes, and associations of pro-amyloidogenic amyloid precursor protein (APP) processing with GluA1 changes. These studies use C57BL/6 mice of both sexes, because gender effects of air pollution on brain functions are understudied. Aim 1: Effects of chronic nPM exposure on hippocampal-dependent learning and cellular correlates. After behavioral testing of chronic nPM exposed mice, hippocampal slices will be analyzed for changes in the AMPA GluA1 vs GluA2 receptor subunits at excitatory Schaffer-collateral hippocampal synapses and in long-term potentiation (LTP). Subcellular fractionation of cortical tissue will address alterations in AMPA receptor trafficking, which regulates synaptic plasticity. The processing of the APP will also be examined in relation to glutamate receptor function. Aim 2: Manipulation of APP processing in male mice with two drugs: a novel gamma secretase modulator (BPN-15606) and with memantine to test the role of endogenous Abeta in cognitive changes and down- regulation of GluA1 during chronic in vivo nPM exposure. Indications of glutamatergic involvement in responses to air pollution nPM are relevant to the epidemiological findings of cognitive impairment because of the role of glutamate receptors in learning and memory and because of AMPA receptor sensitivity to Abeta. Findings will enable deeper analysis of synaptotoxic mechanisms of air pollution, which are needed to identify therapeutic neuroprotection and to adjust permissible environmental limits of ultrafine PM.

PROJECT SUMMARY/ABSTRACT Little is known about the incidence, prevalence, and predictors of Alzheimer's disease (AD) in populations living traditional pre-industrial lifestyles similar to those experienced over human pre-history. This information is critical to determine whether AD is a byproduct of modern environments. Compared to age-matched industrialized populations, Tsimane exhibit: a) delayed atherosclerosis progression over their lifetime; b) low prevalence of diabetes and hypertension; and c) a near absence of atrial fibrillation, stroke and myocardial infarctions. At the same time Tsimane experience high rates of infection and inflammation throughout life. The two major goals of this proposal are to: 1) measure rates of cerebral atrophy and cognitive decline in association with atherosclerotic and inflammatory burden, APOE genotype and schooling, and 2) generate estimates of the prevalence and incidence of all-cause dementia and AD. Our central hypothesis is that compared to Westerners, the low rate of atherosclerosis among Tsimane will be paralleled by a slower rate of cerebral atrophy, and reduced age-related cognitive impairment. We will test the alternative hypothesis that infection and inflammation are associated with accelerated rates of cerebral atrophy and cognitive impairment. To test these predictions we propose the following specific aims, utilizing a panel study design, state-of-the-art bioimaging technology, and a representative sample of 1,310 Tsimane adults aged 40+, which comprises ~85% of the population in that age range: Aim 1 is to conduct longitudinal assessment of cognitive impairment and dementia with measurement of physical activity between assessments; Aim 2 is to conduct anatomic neuroimaging of the brain related to cognitive impairment, AD and other dementias; and Aim 3 is to investigate the epidemiology of brain atrophy, cognitive impairment, AD and other dementias. This research is time-sensitive, as Tsimane are modernizing at an accelerating rate. It may be our last chance to study the natural history of AD, cerebral atrophy and cognitive impairment with a large sample in a population living a subsistence lifestyle, similar to pre- historic populations, with low rates of CVD and high rates of infectious disease. The multi- disciplinary approach will leverage 14 years of Tsimane research, including data on atherosclerosis in four arterial beds, heart disease, infection and inflammation, physical activity level, and cognitive performance. If rates of cerebral atrophy and cognitive impairment are lower among aging Tsimane, despite their high systemic inflammation and limited schooling, those findings will have important implications for our understanding of AD in the US.

? DESCRIPTION (provided by applicant): Prior work has associated air pollution with accelerated atherosclerosis and with the incidence of stroke. Moreover, recent epidemiological studies associate air pollution with accelerated cognitive decline of normal aging. Associations with Alzheimer disease are being studied, supported by postmortem findings that humans exposed to extreme urban pollution have premature brain inflammation and deposits of diffuse amyloid-beta (Abeta). Rodent models support these emerging associations of cognitive dysfunctions with air pollution. After exposure to various urban air pollutants, several rodent genotypes show synaptic changes and increased Abeta, as well as induction of TNFa and other inflammatory responses. Our model uses chronic exposure of mice to nano-scale PM (nPM) derived from urban traffic, which shows greater cytotoxicity than larger PM. Under controlled exposure by duration and dose (nPM density), mice show a 30% decrease of the hippocampal GluA1 AMPA receptor subunit, with little change in other AMPA receptor subunits, or in NMDA receptor subunits, while in vitro studies show that nPM exposure increases excitotoxic damage. Because of its role in fast excitatory transmission and in spatial memory, GluA1 decreases could contribute to cognitive decline in association with air pollutants. Another potential link to brain aging involves interactions of AMPA receptor subunits with Abeta. This proposal addresses key gaps in experimental models for epidemiological associations and postmortem findings: the impact of nPM exposure on hippocampal neuronal physiology associated with cognitive processes, and associations of pro-amyloidogenic amyloid precursor protein (APP) processing with GluA1 changes. These studies use C57BL/6 mice of both sexes, because gender effects of air pollution on brain functions are understudied. Aim 1: Effects of chronic nPM exposure on hippocampal-dependent learning and cellular correlates. After behavioral testing of chronic nPM exposed mice, hippocampal slices will be analyzed for changes in the AMPA GluA1 vs GluA2 receptor subunits at excitatory Schaffer-collateral hippocampal synapses and in long-term potentiation (LTP). Subcellular fractionation of cortical tissue will address alterations in AMPA receptor trafficking, which regulates synaptic plasticity. The processing of the APP will also be examined in relation to glutamate receptor function. Aim 2: Manipulation of APP processing in male mice with two drugs: a novel gamma secretase modulator (BPN-15606) and with memantine to test the role of endogenous Abeta in cognitive changes and down- regulation of GluA1 during chronic in vivo nPM exposure. Indications of glutamatergic involvement in responses to air pollution nPM are relevant to the epidemiological findings of cognitive impairment because of the role of glutamate receptors in learning and memory and because of AMPA receptor sensitivity to Abeta. Findings will enable deeper analysis of synaptotoxic mechanisms of air pollution, which are needed to identify therapeutic neuroprotection and to adjust permissible environmental limits of ultrafine PM.

Project Summary The parent RF1 (1 RF1 AG054442-01) provides new data on brain atrophy and Alzheimer?s disease and Related Dementias (ADRD) in two indigenous Bolivian populations, the Tsimane and the Moseten, with extremely low rates of cardiovascular disease (CVD) but high burdens of infection and inflammation. The aims of this project are: (1) Longitudinal assessment of cognitive impairment and dementia in a low cardiovascular disease (CVD) risk population, Tsimane forager-horticulturalists of lowland Bolivia; (2) Neuroimaging to identify brain atrophy correlates of cognitive impairment, AD and other dementias; (3) Characterization of the prevalence of brain atrophy, cognitive impairment, and dementias in the Tsimane; (4) Collect in- depth data on diet and physical activity; and (5) Examine the interaction of inflammation and APOE genotype on cognitive functioning for individual longitudinal change. ADRD prevalence was estimated during the first 2.5 years of NIA funding. Current results indicate a low prevalence (<5%) of dementia after age 70 with no cases of moderate to severe dementia. The two specific aims of this administrative supplement will determine: (1) whether cognitive impairment and dementia are risk factors for SARS-CoV-2 infection and/or morbidity and mortality, if infected; and (2) if Covid-19 results cognitive and neurological aging impairment. The first aim is necessary for interpreting existing data and achieving the original project goals. it is necessary to determine whether those who develop cognitive impairment dementia are at greater risk for COVID-19 mortality in order to control for a potential downward bias in the estimation of dementia incidence. The second aim presents a new unique opportunity to investigate the original project goal of studying impacts of infection on cognitive decline and ADRD. It will evaluate the impact of COVID-19 infection on cognitive function and ADRD incidence.

  Project 3 Abstract: Caleb Finch, with Todd Morgan and Christian Pike Age-Sex-ApoE Allele Interactions in White Matter Vulnerability to Air Pollution. To reviewers: new text is italicized. Accelerated cognitive aging is strongly associated with air pollution fine- sized particulate matter (PM2.5u) in two recent reports of community-living elderly 1, 2). Besides cognitive deficits and loss of white matter, the WHIMS cohort incurred higher risk of dementia (Project 1). In mouse models, nano-sized PM (nPM) from traffic-related air pollution (TRAP-nPM) is pro- amyloidogenic. In our experimental model, mice receive whole body exposure to TRAP-nPM for 150 hours intermittently during 10 weeks (Core C2). Besides brain-wide inflammatory responses, TRAP-nPM causes selective damage to hippocampal CA1 myelinated neurons (Fig. 4) that are most vulnerable in AD and to cerebrovascular ischemia. Because AD risk is elevated in women, particularly in apoE4 carriers, we propose to study age-sex-ApoE allele interactions in the vulnerability of myelinated pathways to nPM using EFAD mice. We also examine the blood-brain barrier (BBB), which shows greater age-related leakiness in human ApoE4 carriers. New data show that nPM increases activity in a TLR4 inflammatory pathway that mediates post- ischaemic neurodegeneration. We hypothesize that TRAP increases AD risk by TLR4-related inflammatory processes that synergize with CA1-specific hippocampal neurodegenerative mechanisms. Aim 1 (refocused): Age and sex in brain susceptibility of C57BL/6 (B6) mice to nPM. Both sexes of B6 mice will be exposed to nPM at ages 2 mo, 10, and 18 mo, spanning reproductive senescence. We will assay hippocampus mediated learning and hippocampal subregional analysis of myelin degeneration and neuron atrophy. Microglial and TLR4-TNF? pathway responses will be evaluated by immunohistochemistry and Western blots. Cerebrovascular and white matter tract responses to nPM exposure is assessed in Core B2 by in vivo multiphoton imaging for regional CBF and BBB permeability and angiography. DTI-MRI at 80µm isotropic spatial resolution will provide fractional anisotropy maps for white matter connectivity. BBB cellular integrity is assessed by confocal microscopy. Collaboration with Project 4 examines B6 brains for synergies of nPM with chronic cerebral hypoperfusion (CCH). Aim 2: Age-Sex-ApoE allele interactions in EFAD mouse responses to nPM. The Aim 1 parameters of age and sex for hippocampal-mediated learning and neurodegeneration in hippocampal subfields are examined in EFAD mice. Cerebrovascular amyloid and microbleeds will be evaluated in EFAD mice. We hypothesize that nPM will show female bias in accelerating AD changes. Aim 3 (major revisions): Role of TLR4. A new mouse model with inducible macrophage/microglial- specificTLR4 knockout (i-mTLR4-ko) will evaluate TLR4 contributions to nPM-induced myelin degeneration and neurite atrophy. This new model will identify targets of TLR4 pathway components in the effects of TRAP on myelinated pathways.

The Administrative Core supports this interdisciplinary and multi-university team of investigators. The P01 program has three performance sites: the USC University Park Campus (Core C, Projects 3); USC Health Science Campus (Core B, Projects 1 & 4); and University of California, San Diego (Project 2). The proposed program includes an established group of collaborative scientists with expertise in neurobiology of AD, population neuroimaging, mouse brain imaging, brain vascular biology, clinical neurology, cognitive neurosciences, neuropsychology, epidemiology of AD, large-scale air pollution modeling, environmental epidemiology, inhalation exposure assessment and neurotoxicology. The specific aims of the Administrative Core are: 1) Provide the integrated program leadership and coordination; (2) Strengthen existing partnerships, facilitate new collaborations and foster the career development of junior investigators/trainees across USC and participating institutes; (3) Assess scientific progress of the program, through joint group meetings per month, work-in-progress meetings per quarter, and annual meetings of the project-core leaders and key personnel with the External Advisory Committee; (4) Provide fiscal management and administrative services and monitor resources and expenditures; (5) Carry out the joint dissemination of study results to the broader scientific community. The Administrative Core will provide leadership, oversight, planning, and coordination to facilitate scientific integration and synthesis across projects, promote cross-disciplinary interactions, and support multi- site operation of proposed program activities. The collaborative infrastructure of the proposed P01 is the product of the AirPollBrain (PIs: Finch & Chen), a collaborative network funded by USC since 2010. Over the last few years, the AirPollBrain has developed partnerships with several USC-based Centers/Institutes in brain sciences (e.g., ADRC/Memory and Aging Center, Zilkha Neurogenetic Institute, Mark and Mary Stevens Neuroimaging and Informatics Institute; Institute of Developing Mind) and also helped create the new Research Program on Neurological Effects of Environmental Exposure for the latest renewal of Southern California Environmental Health Sciences Center (P30 ES007048-20). The Administrative Core is well positioned to leverage these institutional resources and maximize the impact of the proposed P01 research.

Our overarching goal is to further resolve the neurodegenerative role of traffic-related air pollutants (TRAP), a ubiquitous exposure in urban areas where most older Americans reside. Our recently published epidemiologic data showed strong association between elevated PM2.5 (particulate matter <2.5µm) and increased dementia risk in women >65 years, with a bias for ApoE4 homozygotes (JC Chen in Cacciottolo et al 2017, PMID 28140404). In this same report, experimental studies of female mice from the Finch-Sioutas Labs showed that exposure to nPM (a nano-sized subfraction of TRAP) was pro-amyloidogenic with an ApoE4 bias. Other changes include attrition of hippocampal CA1 neurites and myelin, which model selective damage in AD and cerebral ischemia. In a mouse stroke model, nPM exposure exacerbated cerebral ischemic damage (William Mack: Liu et al 2016, PMID 27071057). Differences by sex and ApoE alleles suggest sources of heterogeneity in human responses to TRAP. We propose four projects: epidemiological studies of two nationwide cohorts of women (Project 1, JC Chen: Women's Health Initiative Memory Studies; WHIMS) and men (Project 2, C Franz and W Kremen: Vietnam Era Twin Study of Aging, VETSA) and two experimental studies of air pollution exposure (Project 3, Finch: mouse models of aging and AD; Project 4 Wm Mack, chronic ceebral hypoperfusion). These projects address a common set of questions: (1) What is the AD risk imposed by TRAP and does the associated risk vary by sex, life stage, and APOE/other alleles? (2) What neurodegenerative changes are induced by TRAP and what is the resulting risk for early cognitive decline of AD? (3) Which brain pathways are most susceptible to TRAP neurotoxicity? (4) Do shared mechanisms (e.g., amyloidogenesis, cerebrovascular damage and hypoperfusion, and neuroinflammation) predispose to premature cognitive decline and an increased AD risk? Two supporting Cores provide population neuroinformatics, neuroimaging of blood-brain-barrier (BBB) and myelinated tracts, large-scale air pollution modeling and epidemiology, inhalation exposure assessment and neurotoxicology. Neuroimaging Core B1 provides human brain imaging harmonized across sites and mediation analyses (Projects1-2). B2 provides high-resolution imaging of BBB and tractography for mouse models (Projects 3-4). Core C Environmental Exposure and Neurotoxicology subcore C1 harmonizes population exposure estimates for WHIMS and VETSA (Projects1-2); C2 provides inhalation exposure of mice for studies of sex and ApoE allele responses (Project 3) and of chronic cerebral hypoperfusion (Project 4); C3 analyzes brain inflammatory protein responses to TRAP. For P01 integration, the Administrative Core builds on the infrastructure of AirPollBrain (led by Finch& Chen), a USC-funded collaborative network since 2010. Results of this program will advance understanding of TRAP contributions to AD risk and accelerated cognitive decline, and provide a rationale for preventive intervention in the environmental neurotoxicology of AD.