Mark Anthony Smith, PhD, Nottingham, FRCPath - US grants

neuropathology; apolipoprotein E; oxidative stress; amyloid proteins; Alzheimer's disease; soma; disease /disorder model; genotype; mitochondria; biomarker; enzyme activity; free radical oxygen; in situ hybridization; genetically modified animals; laboratory mouse; brain; postmortem; immunocytochemistry; human tissue;

Morphology and Immunohistochemistry Core Scientific Abstract An understanding of which cell types within the retina and kidney are affected by diabetes is essential for effective treatment targeting. The cellular localization of specific proteins and cell death markers, and the quantitative analysis of effects of therapies on retinal and renal structure are integral parts of the projects in the current proposal. Specific Aim 1: The purpose of the Morphology and Immunohistochemistry Core is to aid in the execution of anatomically- based experiments by providing: a. technical support for tissue collection and processing, b. expert assistance in evaluation of retinal and renal histopathology, and c. quantify reagents and technical support for immunohistochemistry. Specific Aim 2: The Morphology and Immunohistochemistry Core will facilitate and stimulate research in individual projects by providing expert assistance and special facilities essential for the projects but beyond the current capabilities of investigators. It will also provide quality control and facilitate interaction between investigators, and provide teaching in these technical areas.

[unreadable] DESCRIPTION (provided by applicant): The "Amyloid Cascade Hypothesis" positing amyloid-ft (Aft) as a fundamental driving mechanism in disease pathogenesis is supported by numerous cellular, animal and human studies. However, an "Alternate Amyloid Hypothesis", positing Aft as an antioxidant that occurs secondary to oxidative stress, is an equally valid explanation for the wealth of in vitro and in vivo findings related to Aft. That the "Alternate Amyloid Hypothesis" and the "Amyloid Cascade Hypothesis" are essentially unchallenged by any currently available data is troubling from a scientific perspective and more so from a clinical perspective where current therapeutic efforts are targeted at the removal or limiting of Aft from the brain. The goal of our proposal is to conduct a series of experiments that should critically test both hypotheses to determine the role of Aft as pathogen or protectant. Specifically, we propose to use inducible transfected cell cultures together with state of the art molecular techniques to test the predominant hypothesis in the field. Our Specific Aims, guided by novel preliminary data are as follows: Aim 1) Determine whether pharmacologic inhibition of Aft with BACE inhibitors affects susceptibility to oxidative stress and whether this effect can be reversed ("rescued") by transfection with Aft1-40, Aft1-42, or C99; Aim 2) Determine whether mutations associated with familial Alzheimer disease are associated with alterations in oxidative stress and whether alterations in Aft production are affected by antioxidants or, vice versa, whether oxidative stress is altered by inhibition of Aft; Aim 3) Determine whether Aft is regulated by intracellular redox balance. Completion of the proposed studies will help determine the role of Aft and, more importantly, guide future therapeutic targets. [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): Inappropriate cell cycle control is emerging as an important component in the pathogenesis leading to Alzheimer disease (AD). We and others have shown expression of cell cycle-related proteins in the vulnerable neurons in AD. Evidence that this represents a bona fide mitotic event is proved by the observation that DNA replication is occurring in these cells. The earlier occurrence of cell cycle events compared to other pathologies suggests that a mitotic cell cycle-related mechanism may play a role in AD. Additional supportive evidence for an important role for cell cycle events in disease derives from transgenic animal models. First, in transgenic lines with a high amyloid burden, such as Tg2576, there are increases in cell cycle proteins and chromosome duplication and such changes predate frank amyloid-2 deposition. Second, we recently developed a dedicated transgenic model (CaMKII-MYC) of neuron-specific inducible oncogene driven cell cycle that specifically expresses myc in forebrain neurons. Analysis of such animals shows that MYC expression results in cell cycle re-entry leading to tau phosphorylation, intraneuronal accumulation of amyloid-b, neurodegeneration, and cognitive deficits. The observations from these transgenic lines pinpoint abberant cell cycle control as an early and perhaps pivotal factor in the pathogenesis of AD. We propose to test this notion by inhibiting cell cycle re-entry in both Tg2576 and CaMKII-MYC transgenic models. Using roscovitine to block cell cycle re-entry, we will assess the role of cell cycle re-entry in the pathology (including neuronal degeneration, tau phosphorylation, amyloid-b) and behavioral deficits (such as Morris Water Maze) observed in these mice. At this conclusion of these studies, it is anticipated that we will not only realize the role of cell cycle-mediated events in relation to other aspects of disease pathogenesis but also the potential utility of using cell cycle inhibitory approaches as a therapeutic regimen in AD. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): In previous studies, we and others showed evidence for the aberrant re-expression of a series of cell cycle-related proteins in specific vulnerable neuronal populations in Alzheimer disease (AD). That a mitotic cell cycle-related mechanism may play an important role in disease pathogenesis is highlighted by the earlier occurrence of cell cycle proteins compared to abnormal tau in AD and by their appearance at the very earliest prodrominal phase of disease (i.e., mild cognitive impairment). Furthermore, that cell cycle proteins are representative of a true cell cycle, rather than being an epiphenomena of other processes, is evident from evidence showing that there is a true mitotic alteration that leads to DNA replication (i.e., S phase) in neurons in AD. These findings led us to develop a novel hypothesis that neurodegeneration in AD, like cancer, is a disease of inappropriate cell cycle control. In support of this notion, we found that the expression a powerful cell cycle inducer, MYC, drives primary neurons to re-enter the cell cycle and, moreover, that MYC-induced cell cycle re-entry leads to tau phosphorylation. Based on these in vitro findings, we have recently developed a bitransgenic mouse model that inducibly expresses MYC specifically in forebrain neurons (CaMKII-MYC). In our preliminary analysis of these CaMKII-MYC animals, we found that MYC expression in this animal model: 1) drives neurons to enter the cell cycle;2) causes hyperphosphorylation of tau;3) leads to accumulation of intraneuronal AB;4) results in a "neurodegenerative" (TUNEL) Phenotype;5) leads to gliosis;and 6) causes major cognitive deficits (Y- maze and MWM). Since the aforementioned pathological and behavioral changes are all synonymous with AD, this mouse may be a very useful model of disease pathogenesis. The goal of this proposal is to further delineate the importance of neuronal cell cycle re-entry using this CaMKII-MYC animal model (Aim 1) and examine the effects of neuronal cell cycle re-entry under A2-rich conditions by analysis of a triple transgenic Tg2576/CaMKII-MYC mice (Aim 2). At the conclusion of these studies, we hope to not only have advanced our understanding of fundamental mechanisms that control neuronal cell cycle, particularly as it applies to AD, but also suggest novel therapies that could be manipulated to either prevent initiation of degeneration or stimulate recovery of damaged neurons in neurodegenerative conditions. Alzheimer disease is the leading cause of dementia in the United States. Recent studies have implicated inappropriate cell cycle re-entry in vulnerable neurons as playing a role in the neuronal loss, pathology, and cognitive impairment that are characteristic of the disease. This proposal will help to clarify the role of cell cycle re-entry in Alzheimer disease by characterizing a novel transgenic mouse model of cell cycle re-entry and delineating the effects of cell cycle re-entry in an amyloid-rich environment.

DESCRIPTION (provided by applicant): Title: 38th, 39th, and 40th Annual Meetings of the American Aging Association The 38th Annual Meeting of the American Aging Association (AAA) and 23rd Meeting of the American College of Clinical Gerontology (ACCG) will be held in Scottsdale, AZ, May 29 - June 1, 2009 and the 39th Annual Meeting of the AAA with the 24th Meeting of AACG in Portland, OR, June 4-10, 2010, and the 40th annual meeting from June 3-6, 2011 in Baltimore, MD. The theme for the 2009 meeting is "Integrative Biology, Hormones. Signaling, and Aging," and for the 2010 meeting "Inflammaging: Aging of the Immune System and Age-related Diseases." The theme of the 40th AAA meeting remains to be to be determined. The goal of the annual AAA/AACG meetings is to assemble investigators working in diverse areas of basic and clinical experimental gerontology to review and discuss recent developments in their fields. The AAA/AACG meeting provides a unique forum for presentation of recent advances in aging research and cross-fertilization of different gerontologic disciplines, as well as for recruiting students, recent graduates, and senior scientists into aging research. Based on rate of increase in participation in past meetings, we anticipate that at least 350 scientists, clinicians, students, and laypersons will attend the 2009 meeting and over 400 the 2010 meeting. The aging process is clearly multi-factorial. Therefore, within the themes of the meetings, a wide variety of topics at the forefront of aging research will be presented and discussed. The programs consist of invited and Award lectures presented in Plenary Sessions;additional major sessions consist of 4 to 5 invited presentations, some scheduled in a dual, simultaneous format, and dual sessions of short platform contributed presentations chosen from abstracts submitted by registrants. All sessions incorporate time for questions and discussion. We will also include poster sessions to allow all registrants wishing to present data to do so. Other key elements of each meeting include sessions devoted to enhancement of graduate students'participation in gerontological research, a Trans-Atlantic Symposium funded by Biotechnology and Biological Sciences Research Council of the United Kingdom to support presentation by British scientists and to encourage cross- Atlantic collaboration, and the annual Nathan Shock Center Symposia featuring research presentations by investigators from each of the five NIA-sponsored Nathan Shock Gerontology Research Centers. We request funds to cover costs for providing the venues for these meetings and travel expenses for speakers for the central programs highlighting the following key areas: in 2009- Integrative biology of aging: signaling and coordination from molecules to cells to organisms;Hormones, genes, and longevity;Hormones, skeletal muscle, and neuromuscular junctions;Pituitary and steroid hormones;Nutritional factors;Progress toward negligible senescence;Physiology and genomics of caloric restriction;Hormone effects on the neurobiology of aging;Adipose, insulin resistance, inflammation and atherosclerosis;and Evolutionary issues in aging and age-related disease;in 2010- aging effects on immune system function and related topics. Special efforts will be made to encourage participation of women and minorities. The 2009 meeting will be preceded by a pre- conference workshop, funded by non-NIH sources, on "Protein Quality and Aging," which will, as the title implies, cover a wide variety of topics related to changes in protein synthesis, function, damage, and degradation associated with the aging process. Topics for the 2010 and 2011 Pre-conference workshops remain to be determined. Attachment The graying of the baby boom generation and the progressive increase in life expectancy together are leading to unparalleled expansion of the population over 65 and over 85 years of age, producing a demographic imperative for improvement in the health of the elderly. The annual meetings of the American Aging Association (AAA) serve the important function of helping to inform the public regarding the progress of aging research and the quest for practical means of prolonging the healthy productive portion of human lifespan. In concord with meetings of the American College of Clinical Gerontology (ACCG), the AAA meetings increase awareness and knowledge of gerontology among physicians and other members of the health professions. By bringing together leading investigators working in the various areas of the biology of aging, AAA meetings provide a forum for critical discussion of recent developments and presentation of key findings from ongoing research. Therefore, this meeting is instrumental in stimulating further progress in gerontology and in keeping investigators up-to-date on developments and technologies in experimental gerontology. PUBLIC HEALTH RELEVANCE: Project Narrative Attachment: The graying of the baby boom generation and the progressive increase in life expectancy together are leading to unparalleled expansion of the population over 65 and over 85 years of age, producing a demographic imperative for improvement in the health of the elderly. The annual meetings of the American Aging Association (AAA) serve the important function of helping to inform the public regarding the progress of aging research and the quest for practical means of prolonging the healthy productive portion of human lifespan. In concord with meetings of the American College of Clinical Gerontology (ACCG), the AAA meetings increase awareness and knowledge of gerontology among physicians and other members of the health professions. By bringing together leading investigators working in the various areas of the biology of aging, AAA meetings provide a forum for critical discussion of recent developments and presentation of key findings from ongoing research. Therefore, this meeting is instrumental in stimulating further progress in gerontology and in keeping investigators up-to-date on developments and technologies in experimental gerontology.