The prominent risk factor for Alzheimer's Disease (AD) is aging. Animal studies establish an intimate link between signaling pathways that regulate lifespan and processes that control cellular protein homeostasis, including insulin growth factor 1 receptor (DAF-2/IGF-1R) signaling, dietary restriction (DR) and mitochondria! electron transport chain perturbations. The overall goal of Project 2 is to study cellular aging or senescence in cell-based models that enable us to understand more clearly how aging affects the cell biology that can lead to loss of protein homeostasis control and the onset of AD.
In Aim 1, we hypothesize that aging fundamentally changes the processing of APP into Abeta. We will utilize established fibroblast cell-based aging models and we will develop neuronal cell-based aging models to first demonstrate that aging signaling pathways that control the lifespan of worms and mice also influence cellular lifespan in culture. We will then express APP in these cell-based aging models to discern whether the youthful control of protein homeostasis is lost upon aging of the cells and whether DAF-2/IGF-1R signaling and related pathways influencing aging will extend their lifespan and protect the cells from proteotoxicity, as has been observed in whole animal models including worms and mice. Biochemical and morphological approaches including western blotting, immunofluorescence (IF) and immunoelectron microscopy (IEM) will be used to follow the processing of APP into Abeta, the subcellular appearance of aggregates and their spatial and temporal correlation with proteotoxicity contributing to senescence and cell death.
In Aim 2 we will utilize the aging models to study the membrane trafficking pathways to explore the hypothesis that they function well in young cells yet appear to fail in old cells, in an effort to begin to understand how the expression of APP and its conversion into Abeta becomes toxic with aging in the context of its extensive intracellular processing and trafficking.
Aim 2 is distinguished from Aim 1 by the utilization of numerous known biological perturbants of membrane trafficking pathways in a systematic fashion to discern how these pathways influence the processing of APP into Abeta the subcellular appearance of aggregates and their spatial and temporal correlation with proteotoxicity in the context of aging.

National Institute of Health (NIH)
National Institute on Aging (NIA)
Research Program Projects (P01)
Project #
Application #
Study Section
Special Emphasis Panel (ZAG1-ZIJ-8)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Scripps Research Institute
La Jolla
United States
Zip Code
Roth, Daniela Martino; Hutt, Darren M; Tong, Jiansong et al. (2014) Modulation of the maladaptive stress response to manage diseases of protein folding. PLoS Biol 12:e1001998
Park, Sung Kyu Robin; Aslanian, Aaron; McClatchy, Daniel B et al. (2014) Census 2: isobaric labeling data analysis. Bioinformatics 30:2208-9
Greiner, Erin R; Kelly, Jeffery W; Palhano, Fernando L (2014) Immunoprecipitation of amyloid fibrils by the use of an antibody that recognizes a generic epitope common to amyloid fibrils. PLoS One 9:e105433
Bamberger, Casimir; Pankow, Sandra; Park, Sung Kyu Robin et al. (2014) Interference-free proteome quantification with MS/MS-based isobaric isotopologue detection. J Proteome Res 13:1494-501
Tsigelny, Igor F; Sharikov, Yuriy; Kouznetsova, Valentina L et al. (2014) Structural diversity of Alzheimer's disease amyloid-* dimers and their role in oligomerization and fibril formation. J Alzheimers Dis 39:583-600
Amschl, David; Neddens, Jorg; Havas, Daniel et al. (2013) Time course and progression of wild type ýý-synuclein accumulation in a transgenic mouse model. BMC Neurosci 14:6
Kim, Changyoun; Ho, Dong-Hwan; Suk, Ji-Eun et al. (2013) Neuron-released oligomeric *-synuclein is an endogenous agonist of TLR2 for paracrine activation of microglia. Nat Commun 4:1562
Yates 3rd, John R (2013) The revolution and evolution of shotgun proteomics for large-scale proteome analysis. J Am Chem Soc 135:1629-40
Toyama, Brandon H; Savas, Jeffrey N; Park, Sung Kyu et al. (2013) Identification of long-lived proteins reveals exceptional stability of essential cellular structures. Cell 154:971-82
Dusaban, Stephanie S; Purcell, Nicole H; Rockenstein, Edward et al. (2013) Phospholipase C epsilon links G protein-coupled receptor activation to inflammatory astrocytic responses. Proc Natl Acad Sci U S A 110:3609-14

Showing the most recent 10 out of 41 publications