A Human Pluripotent Stem Cell-Based Screen for Smooth Muscle Cell Differentiation and Maturation Identifies Inhibitors of Intimal Hyperplasia

Stem Cell Reports. 2019 Jun 11;12(6):1269-1281. doi: 10.1016/j.stemcr.2019.04.013. Epub 2019 May 9.

Abstract

Contractile to synthetic phenotypic switching of smooth muscle cells (SMCs) contributes to stenosis in vascular disease and vascular transplants. To generate more contractile SMCs, we performed a high-throughput differentiation screen using a MYH11-NLuc-tdTomato human embryonic stem cell reporter cell line. We identified RepSox as a factor that promotes differentiation of MYH11-positive cells by promoting NOTCH signaling. RepSox induces SMCs to exhibit a more contractile phenotype than SMCs generated using PDGF-BB and TGF-β1, two factors previously used for SMC differentiation but which also cause intimal hyperplasia. In addition, RepSox inhibited intimal hyperplasia caused by contractile to synthetic phenotypic switching of SMCs in a rat balloon injury model. Thus, in addition to providing more contractile SMCs that could prove useful for constructing artificial blood vessels, this study suggests a strategy for identifying drugs for inhibiting intimal hyperplasia that act by driving contractile differentiation rather than inhibiting proliferation non-specifically.

Keywords: MYH11-NLuc-tdTomato reporter cell line; NOTCH; RepSox; contractile smooth muscle cells; differentiation; high-throughput screen; intima hyperplasia; maturation; pluripotent stem cells; restenosis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Becaplermin / metabolism
  • Cell Differentiation*
  • Disease Models, Animal
  • Humans
  • Hyperplasia
  • Muscle, Smooth, Vascular / metabolism*
  • Muscle, Smooth, Vascular / pathology
  • Myocytes, Smooth Muscle / metabolism*
  • Myocytes, Smooth Muscle / pathology
  • Myosin Heavy Chains / metabolism
  • Pluripotent Stem Cells / metabolism*
  • Pluripotent Stem Cells / pathology
  • Rats
  • Transforming Growth Factor beta1 / metabolism
  • Tunica Intima / metabolism*
  • Tunica Intima / pathology

Substances

  • MYH11 protein, human
  • TGFB1 protein, human
  • Transforming Growth Factor beta1
  • Becaplermin
  • Myosin Heavy Chains