instmctions): (Project 5, Apte) Versican Proteolysis and Regulation of Vascular Smooth Muscle Function -Versican, an extracellular matrix (ECM) proteoglycan, is a prominent product of vascular smooth muscle cells (VSMCs) and the developing heart. Its proteolytic turnover by ADAMTS metalloproteases is a major mechanism underlying cardiac outflow tract and myocardial development. In contrast, the role of ADAMTS-mediated versican proteolysis in the vascular wall is poorly understood. Our preliminary data demonstrate that ADAMTSI, ADAMTS5 and ADAMTS9 are expressed by embryonic and adult VSMCs, and we propose a cooperative mode of action of these proteases in versican turnover. Preliminary studies identified aortic wall anomalies associated with versican accumulation in Adamts9+/- mice . We found that ADAMTS proteolytic activity influenced the volume of the hyaluronan-based pericellular matrix of cultured fibroblasts, of which versican is a critical component. Associated with this change, we saw enhanced TGF-beta signaling and acquisition of a myofibroblastic phenotype. The hypothesis underlying the proposal is that ADAMTS proteases work cooperatively in VSMCs to regulate the structural and pericellular matrix. Through this physiological function, they are proposed to influence VSMC behavior and the development and integrity of the vasculature. The hypothesis will be addressed by taking a combined genetic and cell biology approach. Using mice deficient In Adamtsi, Adamts5, and Adamts9, we will use combinatorial genetics in aim 1 to define the individual and cooperative roles of these genes in the vasculature. Using VSMCs isolated from these mouse mutants, we will determine the role of ADAMTS proteases in influencing VSMC function in aim 2. We will complement genetic experiments with approaches that use wild-type VSMCs in conjunction with chemical ADAMTS inhibition, as well as gain-of-function strategies (increased expression of versican and ADAMTS) to comprehensively define the underlying mechanisms. Since versican in the pericellular matrix is complexed to hyaluronan (HA), we will undertake quantitative analysis of the effect of ADAMTS proteases on the dynamics of the versican-HA glycocalyx using resources available in Core B.

Public Health Relevance

These experiments address a critical area of vascular biology and medicine, with potential relevance for developmental defects, aneurysms, re-stenosis and atherosclerosis. The research is expected to provide new candidate genes for genetic predisposition to these conditions, and the mechanisms that are uncovered are likely to offer potentially new therapeutic approaches.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL107147-04
Application #
8669090
Study Section
Special Emphasis Panel (ZHL1-CSR-H)
Project Start
Project End
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
4
Fiscal Year
2014
Total Cost
$354,303
Indirect Cost
$128,248
Name
Cleveland Clinic Lerner
Department
Type
DUNS #
135781701
City
Cleveland
State
OH
Country
United States
Zip Code
44195
Kim, Yeojung; West, Gail A; Ray, Greeshma et al. (2017) Layilin is critical for mediating hyaluronan 35kDa-induced intestinal epithelial tight junction protein ZO-1 in vitro and in vivo. Matrix Biol :
Kessler, Sean P; Obery, Dana R; Nickerson, Kourtney P et al. (2017) Multifunctional Role of 35 Kilodalton Hyaluronan in Promoting Defense of the Intestinal Epithelium. J Histochem Cytochem :22155417746775
Saikia, Paramananda; Bellos, Damien; McMullen, Megan R et al. (2017) MicroRNA 181b-3p and its target importin ?5 regulate toll-like receptor 4 signaling in Kupffer cells and liver injury in mice in response to ethanol. Hepatology 66:602-615
Zahreddine, Hiba Ahmad; Culjkovic-Kraljacic, Biljana; Emond, Audrey et al. (2017) The eukaryotic translation initiation factor eIF4E harnesses hyaluronan production to drive its malignant activity. Elife 6:
Ghatak, Shibnath; Markwald, Roger R; Hascall, Vincent C et al. (2017) Transforming growth factor ?1 (TGF?1) regulates CD44V6 expression and activity through extracellular signal-regulated kinase (ERK)-induced EGR1 in pulmonary fibrogenic fibroblasts. J Biol Chem 292:10465-10489
Petrey, Aaron C; de la Motte, Carol A (2017) The extracellular matrix in IBD: a dynamic mediator of inflammation. Curr Opin Gastroenterol 33:234-238
Ghatak, Shibnath; Hascall, Vincent C; Markwald, Roger R et al. (2017) Transforming growth factor ?1 (TGF?1)-induced CD44V6-NOX4 signaling in pathogenesis of idiopathic pulmonary fibrosis. J Biol Chem 292:10490-10519
Asano, Keiichi; Nelson, Courtney M; Nandadasa, Sumeda et al. (2017) Stromal Versican Regulates Tumor Growth by Promoting Angiogenesis. Sci Rep 7:17225
Hendee, Kathryn; Wang, Lauren Weiping; Reis, Linda M et al. (2017) Identification and functional analysis of an ADAMTSL1 variant associated with a complex phenotype including congenital glaucoma, craniofacial, and other systemic features in a three-generation human pedigree. Hum Mutat 38:1485-1490
Stober, Vandy P; Johnson, Collin G; Majors, Alana et al. (2017) TNF-stimulated gene 6 promotes formation of hyaluronan-inter-?-inhibitor heavy chain complexes necessary for ozone-induced airway hyperresponsiveness. J Biol Chem 292:20845-20858

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