title: NOTCH1 regulates matrix gla protein and calcification gene networks in human valve endothelium.
abstract: Valvular and vascular calcification are common causes of cardiovascular morbidity and mortality. Developing effective treatments requires understanding the molecular underpinnings of these processes. Shear stress is thought to play a role in inhibiting calcification. Furthermore, NOTCH1 regulates vascular and valvular endothelium, and human mutations in NOTCH1 can cause calcific aortic valve disease. Here, we determined the genome-wide impact of altering shear stress and NOTCH signaling on human aortic valve endothelium. mRNA-sequencing of primary human aortic valve endothelial cells (HAVECs) with or without knockdown of NOTCH1, in the presence or absence of shear stress, revealed NOTCH1-dependency of the atherosclerosis-related gene connexin 40 (GJA5), and numerous repressors of endochondral ossification. Among these, matrix gla protein (MGP) is highly expressed in aortic valve and vasculature, and inhibits soft tissue calcification by sequestering bone morphogenetic proteins (BMPs). Altering NOTCH1 levels affected MGP mRNA and protein in HAVECs. Furthermore, shear stress activated NOTCH signaling and MGP in a NOTCH1-dependent manner. NOTCH1 positively regulated endothelial MGP in vivo through specific binding motifs upstream of MGP. Our studies suggest that shear stress activates NOTCH1 in primary human aortic valve endothelial cells leading to downregulation of osteoblast-like gene networks that play a role in tissue calcification.
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: Mark P
authors: MP
authors: White
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: Christina V
authors: CV
authors: Theodoris
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: Lei
authors: L
authors: Liu
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: William J
authors: WJ
authors: Collins
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: Kathleen W
authors: KW
authors: Blue
authors: University of Colorado Denver School of Medicine, Aurora, CO, USA.
authors: Joon Ho
authors: JH
authors: Lee
authors: University of Colorado Denver School of Medicine, Aurora, CO, USA.
authors: Xianzhong
authors: X
authors: Meng
authors: Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, USA.
authors: Robert C
authors: RC
authors: Robbins
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA.
authors: Kathryn N
authors: KN
authors: Ivey
authors: Gladstone Institute of Cardiovascular Disease and University of California, San Francisco, USA. Electronic address: dsrivastava@gladstone.ucsf.edu.
authors: Deepak
authors: D
authors: Srivastava
mesh_terms: D001021:Aortic Valve / Q000473:pathology*; D001024:Aortic Valve Stenosis / Q000235:genetics* / Q000473:pathology; D002114:Calcinosis / Q000235:genetics* / Q000473:pathology; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D016000:Cluster Analysis; D004247:DNA / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D015894:Genome, Human; D006801:Humans; D011485:Protein Binding; D051881:Receptor, Notch1 / Q000378:metabolism*; D012212:Rheology; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000235:genetics; D013314:Stress, Mechanical; D000097568:Matrix Gla Protein
chemical_list: D002135:Calcium-Binding Proteins; D004247:DNA; D016326:Extracellular Matrix Proteins; D051881:Receptor, Notch1; D000097568:Matrix Gla Protein
keywords: Matrix gla protein; NOTCH signaling; NOTCH1; Valve calcification; Valve endothelium