CREG ameliorates embryonic stem cell differentiation into smooth muscle cells by modulation of TGF-β expression

• Published in: Differentiation (London) Vol. 125; pp. 9 - 17
• Main Authors: Peng, Chengfei, Shao, Xiaoping, Tian, Xiaoxiang, Li, Yang, Liu, Dan, Yan, Chenghui, Han, Yaling
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01-05-2022; Elsevier Science Ltd
• Subjects: Actin; Angiotensin; Angiotensin II; Calponin; Cell culture; Cell differentiation; Cell Differentiation - genetics; Cells, Cultured; Contraction; CREG; Embryonic stem cells; Embryonic Stem Cells - metabolism; Muscle contraction; Myocytes, Smooth Muscle - metabolism; Repressor Proteins - genetics; Repressor Proteins - metabolism; Smad2 protein; Smooth muscle; Smooth muscle cells; Stem cells; TGF-β; Transfection; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - metabolism; TGF-β; Embryonic stem cells; Smooth muscle cells; CREG; Contraction
• ISSN: 0301-4681; 1432-0436
• DOI: 10.1016/j.diff.2022.03.001

Vascular smooth muscle cell (SMCs) differentiation is critical for cardiovascular development, but the mechanisms remain largely unknown. The overall aim of this study was to investigate the functional impact and mechanism of cellular repressor of E1A-stimulated genes (CREG) in SMC differentiation. Two embryonic stem cell (ESC) models were generated (1) the overexpression of CREG (CREG-OE), by transfection with Pcreg-IRECS2-EGFP vector, and (2) the knockout of CREG, by transfection with CREG shRNA (CREG-KO). Interesting, SMC-marker levels (SM α-actin, SM22, Calponin, and SM-MHC) dramatically increased in CREG-OE ESCs into the SMC while significantly decreased in CREG-KO ESCs during differentiation. After 14 days, and calcium ion concentrations in angiotensin II-stimulated embryoid bodies were increased in CREG-OE ESCs but reduced in CREG-KO ESCs. Consistently, the contractile capacity of SMC from CREG-OE ESC was increased, while the contractile capacity of SMC CREG1 from CREG-KO ESCs was significantly reduced. Furthermore, we demonstrated that CREG promotes differentiation of ESCs to SMCs and maturation of their function through the transforming growth factor-β -smad2/3 pathway.