Alphabeta generation occurs via serial cleavage of APP by p- and gamma-secretase, but is precluded via serial cleavage by alpha- and gamma-secretase. Project 1 (Selkoe/Wolfe) has generated evidence for an alpha/gamma-secretase complex, which we have termed the "sheddasome". In this project, we will explore factors that can modulate the sheddasome either pharmacologically (using novel gamma-secretase modulators [GSM]), or genetically (using novel late-onset Alzheimer's disease [LOAD] mutations in ADAMVO which we discovered during the current PPG period). In our first set of aims, we will carry out studies of a novel series of highly potent, APP-specific GSMs. These GSMs are aryl 2-aminothiazole GSMs that bind directly to the gamma-secretase complex, decreasing AP42 and AP40 levels and increasing APas and AP37 levels. This project will serve to leverage an independent ongoing project on these GSMs supported by the NIH Blueprint Neurotherapeutics Network (NIH-BNN). While we receive no funding from the NIH-BNN, we co-developed these GSM'and serve as close collaborators and Lead Development Team members for the project. As part of Aim 1, in collaboration with Project 1, we will test for the effects of these novel GSMs on gamma-secretase preparations, including gamma-secretase- enriched membranes and sheddasome complexes. We will also test whether the GSMs allosterically impact PS1 conformation in the gamma-secretase complex in collaboration with Project 3 (Berezovska/Hyman). Finally, we will test for the potency and substrate selectivity of these GSMs in collaboration with Project 4 (Kovacs). In the second set of aims, we will characterize two rare LOAD missense mutations in the prodomain oi ADAMIO, which we have already shown to tightly co-segregate with LOAD in 7 families (age of onset ~70 yr). We have also shown that both of these mutations significantly attenuate alpha-secretase activity and elevate alphabeta levels (relative to wild-type) in vitro and in vivo. We have already generated transgenic mice overexpressing either wild-type (WT) or mutant (Q170H;R181G; dominant-negative) forms of ADAMIO. We will use these animal models to identify physiologic ADAM 10 substrates, and characterize their serial cleavage by gamma-secretase (with Project 4). In collaboration with Project 1, we will test whether the LOAD and dominant negative mutations in ADAMIO affect the interaction of ADAM10 with gamma-secretase in the sheddasome.

National Institute of Health (NIH)
Research Program Projects (P01)
Project #
Application #
Study Section
Special Emphasis Panel (ZAG1)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Brigham and Women's Hospital
United States
Zip Code
Holmes, Oliver; Paturi, Swetha; Wolfe, Michael S et al. (2014) Functional analysis and purification of a Pen-2 fusion protein for ?-secretase structural studies. J Neurochem 131:94-100
McFarland, Karen N; Huizenga, Megan N; Darnell, Shayna B et al. (2014) MeCP2: a novel Huntingtin interactor. Hum Mol Genet 23:1036-44
Li, Yi; Lu, Stephen Hsueh-Jeng; Tsai, Ching-Ju et al. (2014) Structural interactions between inhibitor and substrate docking sites give insight into mechanisms of human PS1 complexes. Structure 22:125-35
Choi, Se Hoon; Kim, Young Hye; Hebisch, Matthias et al. (2014) A three-dimensional human neural cell culture model of Alzheimer's disease. Nature 515:274-8
Bolduc, David M; Wolfe, Michael S (2014) Structure of nicastrin unveils secrets of ?-secretase. Proc Natl Acad Sci U S A 111:14643-4
Winslow, Ashley R; Moussaud, Simon; Zhu, Liya et al. (2014) Convergence of pathology in dementia with Lewy bodies and Alzheimer's disease: a role for the novel interaction of alpha-synuclein and presenilin 1 in disease. Brain 137:1958-70
Holmes, Oliver; Paturi, Swetha; Selkoe, Dennis J et al. (2014) Pen-2 is essential for ?-secretase complex stability and trafficking but partially dispensable for endoproteolysis. Biochemistry 53:4393-406
Kim, Doo Yeon; Wertz, Mary H; Gautam, Vivek et al. (2014) The E280A presenilin mutation reduces voltage-gated sodium channel levels in neuronal cells. Neurodegener Dis 13:64-8
Wahlster, Lara; Arimon, Muriel; Nasser-Ghodsi, Navine et al. (2013) Presenilin-1 adopts pathogenic conformation in normal aging and in sporadic Alzheimer's disease. Acta Neuropathol 125:187-99
Li, Xuejing; Uemura, Kengo; Hashimoto, Tadafumi et al. (2013) Neuronal activity and secreted amyloid ? lead to altered amyloid ? precursor protein and presenilin 1 interactions. Neurobiol Dis 50:127-34

Showing the most recent 10 out of 95 publications