Small molecule-mediated TGF-β type II receptor degradation promotes cardiomyogenesis in embryonic stem cells

Cell Stem Cell. 2012 Aug 3;11(2):242-52. doi: 10.1016/j.stem.2012.04.025.

Abstract

The cellular signals controlling the formation of cardiomyocytes, vascular smooth muscle, and endothelial cells from stem cell-derived mesoderm are poorly understood. To identify these signals, a mouse embryonic stem cell (ESC)-based differentiation assay was screened against a small molecule library resulting in a 1,4-dihydropyridine inducer of type II TGF-β receptor (TGFBR2) degradation-1 (ITD-1). ITD analogs enhanced proteasomal degradation of TGFBR2, effectively clearing the receptor from the cell surface and selectively inhibiting intracellular signaling (IC(50) ~0.4-0.8 μM). ITD-1 was used to evaluate TGF-β involvement in mesoderm formation and cardiopoietic differentiation, which occur sequentially during early development, revealing an essential role in both processes in ESC cultures. ITD-1 selectively enhanced the differentiation of uncommitted mesoderm to cardiomyocytes, but not to vascular smooth muscle and endothelial cells. ITD-1 is a highly selective TGF-β inhibitor and reveals an unexpected role for TGF-β signaling in controlling cardiomyocyte differentiation from multipotent cardiovascular precursors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • Cells, Cultured
  • Dihydropyridines / chemistry
  • Dihydropyridines / pharmacology*
  • Dose-Response Relationship, Drug
  • Down-Regulation / drug effects*
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / drug effects*
  • Embryonic Stem Cells / metabolism
  • Epidermal Growth Factor / deficiency
  • Epidermal Growth Factor / metabolism
  • HEK293 Cells
  • Humans
  • Membrane Glycoproteins / deficiency
  • Membrane Glycoproteins / metabolism
  • Mice
  • Mice, Knockout
  • Molecular Weight
  • Myocytes, Cardiac / cytology
  • Myocytes, Cardiac / drug effects*
  • Myocytes, Cardiac / metabolism
  • Neoplasm Proteins / deficiency
  • Neoplasm Proteins / metabolism
  • Protein-Serine-Threonine Kinases / deficiency*
  • Protein-Serine-Threonine Kinases / metabolism*
  • Proteolysis / drug effects*
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta / deficiency*
  • Receptors, Transforming Growth Factor beta / metabolism*
  • Structure-Activity Relationship

Substances

  • Dihydropyridines
  • Membrane Glycoproteins
  • Neoplasm Proteins
  • Receptors, Transforming Growth Factor beta
  • Tdgf1 protein, mouse
  • Epidermal Growth Factor
  • 1,4-dihydropyridine
  • Protein-Serine-Threonine Kinases
  • Receptor, Transforming Growth Factor-beta Type II