Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is a stroke disorder caused by mutations in Notch3. Hallmark pathological features of CADASIL include: rarefication of the extracellular matrix, granular osmiophilic material (GOM) deposition, and Notch3 protein accumulation. Identification of molecular cascades that lead to these abnormalities will potentially lead to therapies which can prevent progression of this debilitating disease. Normal Notch signaling requires trans-endocytosis of the Notch3 ectodomain, resulting in movement of Notch from one cell to another. This process reduces Notch3 levels. We propose that defects in trans-endocytosis result all of the hallmark pathological features of CADASIL. We have recently identified a family of proteins that interacts with Notch3 and participate in trans-endocytosis of Notch3. Preliminary data demonstrates specific physical interactions between Notch3 and LRP1, a protein known for its endocytic function. Our data indicate that LRP1 plays an important role in trans- endocytosis of Notch3, which potentiates Notch3 function. Based on these findings, we suggest the following hypothesis: mutant Notch3 in CADASIL dysfunctionally binds to LRP1, leading to LRP1 malfunction;decreased LRP1 results in inhibition of endocytosis of critical extracellular proteins, including Notch3. We propose to test this hypothesis in three specific aims. First, we will determine at the molecular level whether mutant Notch3 proteins interact differently with LRP1 (compared to WT Notch3). Second, we will determine in cell cultures whether mutant Notch3 inhibits LRP1 dependent endocytosis. Third, we will test our overall hypothesis by examining mice with tissue-specific inactivation of LRP1 to test whether LRP1 is a true target of mutant Notch3 in vivo. These studies may lead to important directions in the treatment of CADASIL, since our main hypothesis indicates that targeting the interaction between mutant Notch3 and LRP1 may slow the disease process. In addition, recent evidence shows that LRP1 participates in vascular dysfunction after stroke;our studies may thus offer additional insights into the mechanisms of how LRP1 in the brain regulates vascular homeostasis. In lay termninology, this proposal will define the precise mechanism of how Notch proteins can be regulated by LRP1. We will determine if CADASIL Notch3 mutants influence LRP1 function, and whether LRP1 dysfunction results in the vascular pathology seen in CADASIL patients.

Public Health Relevance

CADASIL is a prototype inherited stroke disorder caused by Notch3 mutations and protein accumulation. Investigation of how Notch3 accumulates in CADASIL cells may offer clues to the molecular pathways governing this disorder. In addition, we intend to study the regulation of wild type Notch levels. Since Notch signaling is responsible for blood vessel growth and normal development, these studies may impact a broad range of disorders including cancer, heart disease, and stroke. LRP1 plays novel roles in regulation of vasculature after injury;therefore, these studies promise to shed light on mechanisms of vascular homeostasis in the brain.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
3R01NS062816-04S1
Application #
8524606
Study Section
Brain Injury and Neurovascular Pathologies Study Section (BINP)
Program Officer
Corriveau, Roderick A
Project Start
2009-08-01
Project End
2014-07-31
Budget Start
2012-08-01
Budget End
2013-07-31
Support Year
4
Fiscal Year
2012
Total Cost
$18,000
Indirect Cost
$6,424
Name
University of Michigan Ann Arbor
Department
Neurology
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Zhang, Xiaojie; Lee, Soo Jung; Young, Marian F et al. (2015) The small leucine-rich proteoglycan BGN accumulates in CADASIL and binds to NOTCH3. Transl Stroke Res 6:148-55
Keep, Richard F; Wang, Michael M; Xiang, Jianming et al. (2014) Full steam ahead with remote ischemic conditioning for stroke. Transl Stroke Res 5:535-7
Zhang, Xiaojie; Lee, Soo Jung; Young, Kelly Z et al. (2014) Latent NOTCH3 epitopes unmasked in CADASIL and regulated by protein redox state. Brain Res 1583:230-6
Lee, Soo Jung; Zhang, Xiaojie; Wang, Michael M (2014) Vascular accumulation of the small leucine-rich proteoglycan decorin in CADASIL. Neuroreport 25:1059-63
Dong, Hairong; Ding, Haixia; Young, Kelly et al. (2013) Advanced intimal hyperplasia without luminal narrowing of leptomeningeal arteries in CADASIL. Stroke 44:1456-8
Wang, Michael M; Lee, Soo Jung; Kim, Jisu et al. (2013) ABO blood antigens define human cerebral endothelial diversity. Neuroreport 24:79-83
Zhang, Xiaojie; Meng, He; Wang, Michael M (2013) Collagen represses canonical Notch signaling and binds to Notch ectodomain. Int J Biochem Cell Biol 45:1274-80
Meng, He; Zhang, Xiaojie; Lee, Soo Jung et al. (2013) Von Willebrand factor inhibits mature smooth muscle gene expression through impairment of Notch signaling. PLoS One 8:e75808
Wang, Michael M; Xi, Guohua; Keep, Richard F (2013) Should the STAIR criteria be modified for preconditioning studies? Transl Stroke Res 4:3-14
Frieler, Ryan A; Ray, Jessica J; Meng, He et al. (2012) Myeloid mineralocorticoid receptor during experimental ischemic stroke: effects of model and sex. J Am Heart Assoc 1:e002584

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