18 GOALS FORFELLOWSHIPTRAININGANDCAREER Name (Last,first,middle initial) Linford, Nancy J INSTITUTION MENTOR Harvey Mudd College University of Washington Dr Dan Dorsa INSTITUTIONJCOMPANY SUPERVISOR/EMPLOYER My basic research interests lie in the discovery of cellular mechanisms associated with human aging with the goal of discovering and developing therapies for increasing quality of life in the elderly and extending human healthspan It is my long term goal to pursue these research lines as an independent investigator in a highly collaborative university environment In this fellowship I plan to explore the role of mitochondrial oxidative stress in aging I will learn techniques including the culture of cardiac myocytes, assays of cell function and physiology by flow cytometry, cDNA microarray analysis and informatics, and real time RT-PCR With this I hope to broaden my approach from the study of individual intracellular markers to include assays that can simultaneously measure multiple factors and give a more global picture of cellular response to a stimulus I will also have the opportunity to use a quantitative approach to traditional assays of mRNA and protein expression by leaming quantitative immunofluorescence analysis and real time RT-PCR This fellowship gives me the opportunity to use cutting edge techniques in a laboratory that is well established in studies of aging at an institution that has the highest quality instrumentation and infrastructure as well as world-renowned researchers that will be an invaluable advantage for doing truly novel research and develolin I mrselfasan investilator tiPl,Ft'], 19 NAMEANDDEGREE(S) Dr Peter Rabinovitch, MD, PhD (Dr George Martin, MD Co-sponsor) 20 POSITION/RANK Professor, Department of Pathology (PR and GM) 21 RESEARCHINTERESTS/AREAS Molecular aspects of aging and cancer development 22 DESCRIPTION(Do notexceedspaceprovided) The oxidative damage theory of aging proposes that oxygen free radicals are responsible for the cellular changes associated with aging The majority of reactive oxygen species (ROS) are produced in the mitochondria as a byproduct of mitochondrial energy production To test this theory in mammals, Drs George Martin and Peter Rabinovitch have created a transgenic mouse model overexpressing catalase, an enzyme that prevents the formation of oxygen free radicals from hydrogen peroxide Using a beta actin promoter, the catalase was expressed chiefly in muscle and heart Targeting the catalase to mitochondria (by addition of a mitochondrial localization signal- mCAT), but not to nuclei or peroxisomes, results in a 20% increase in average lifespan and a reduction in cardiac pathology during aging (ms in preparation) This is the first genetic model in mammals to demonstrate the link between mitochondrial oxidative stress and aging The proposed experiments will investigate the molecular connections between reduced mitochondrial oxygen free radical formation and reduced cardiac pathology in this aging model We hope to establish a clear mechanism for this effect and to uncover potentially novel intracellular mediators of aging First, the response of cultured cardiac myocytes to cellular stressors that are believed to act through formation of mitochondrial oxidative stress will be analyzed to confirm at a cellular level that the overexpression of mCAT reduces intracellular signaling associated with oxidative stress Next, mitochondrial energy turnover will be analyzed and determine whether the increase in lifespan is correlated with a reduction in age-related decline of mitochonddal function Thirdly, cDNA microarrays will be used to determine the profile of age-related gene expression changes in cultured cardiac myocytes from mCAT vs control mice These experiments will provide novel insight into the intracellular changes that correlate the overexpression of catalase in mitochondria with life extension and reduction of cardiac pathology PHS416-1 (Rev 12/98) FormPage 2 BB CC Individual NRSA Application Table of Contents ========================================Section End===========================================
| Qin, Li-Xuan; Beyer, Richard P; Hudson, Francesca N et al. (2006) Evaluation of methods for oligonucleotide array data via quantitative real-time PCR. BMC Bioinformatics 7:23 |
