The long term goal of this line of investigation is to identify the factors that can provide inherent protection to the endothelium during the initial stages of atherosclerosis and inflammation. In this proposal, we will focus on NOTCH1, a cell surface receptor and transcription factor that, based on preliminary data, provides an anti-inflammatory phenotype to quiescent (homeostatic) endothelium. NOTCH1 is constitutively expressed by the adult endothelium of mouse and human vessels in vivo. Reduction of NOTCH1 transcripts in human endothelium in vitro or genetic inactivation of Notch1 in mice triggers an inflammatory response in the absence of any additional insult. Exposure of endothelial cells to Western diet, oxidized phospholipids (Ox- PAPC), as well as inflammatory cytokines, results in a rapid reduction in endogenous NOTCH1 (together with its target genes). Using microarray analyses of endothelial cells from 147 human donors, we observed differences in basal and Ox-PAPC treated levels of NOTCH1 and identified a locus that was associated with the response of endothelial cells to NOTCH1 by Ox-PAPC. This same locus was also associated with HDL levels in a large scale GWAS including 100,000 humans. Studies in this project will test the hypothesis that reduction of NOTCH1 by circulating lipids contributes to the prolonged inflammation typical of atherosclerosis lesions. In fact, reduction of NOTCH1 levels in an athero-susceptible background (Apoe or Ldlr null mice) promotes acceleration of atherosclerosis and results in larger lesions. Furthermore, genetic inactivation of Notch1 in the endothelium of adult mice leads to leukocyte infiltration, detachment and loss of endothelial cells from the intima. In addition and consistent with a role in endothelial homeostasis and suppression of inflammation, using a ChIP approach we found that NOTCH1 regulates tristetraprolin, an RNA binding protein that targets to AREs sequences in the 3'UTR causing destabilization of mRNAs encoding a cohort of inflammatory cytokines. The central hypothesis of this application is that NOTCH1 in the endothelium is essential to maintain an anti-inflammatory interface between blood and tissue. To test this hypothesis, we propose two specific aims: 1. To determine the mechanism(s) that control NOTCH1 expression and function in adult arterial endothelium; and 2. To identify the molecular pathways by which NOTCH1 maintains an anti-inflammatory status in the endothelium of quiescent arteries.

Public Health Relevance

Inflammation of blood vessels is a well-recognized predisposing factor that triggers cardiovascular disease. Experiments outlined in this proposal aim at identifying the endogenous genes that prevent inflammation in surface of blood vessels with the ultimate goal of suppressing the initiation of atherosclerosis.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL130290-01
Application #
9007956
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Gao, Yunling
Project Start
2015-12-01
Project End
2019-11-30
Budget Start
2015-12-01
Budget End
2016-11-30
Support Year
1
Fiscal Year
2016
Total Cost
$385,000
Indirect Cost
$135,000
Name
University of California Los Angeles
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
McDonald, Austin I; Shirali, Aditya S; Aragón, Raquel et al. (2018) Endothelial Regeneration of Large Vessels Is a Biphasic Process Driven by Local Cells with Distinct Proliferative Capacities. Cell Stem Cell 23:210-225.e6
Mack, Julia J; Mosqueiro, Thiago S; Archer, Brian J et al. (2017) NOTCH1 is a mechanosensor in adult arteries. Nat Commun 8:1620
Sunshine, Hannah; Iruela-Arispe, Maria Luisa (2017) Membrane lipids and cell signaling. Curr Opin Lipidol 28:408-413
Shirali, Aditya S; McDonald, Austin I; Mack, Julia J et al. (2016) Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model. J Vis Exp :
Uebelhoer, Melanie; Iruela-Arispe, M Luisa (2016) Cross-talk between signaling and metabolism in the vasculature. Vascul Pharmacol 83:4-9
He, Huanhuan; Mack, Julia J; Güç, Esra et al. (2016) Perivascular Macrophages Limit Permeability. Arterioscler Thromb Vasc Biol 36:2203-2212