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Hum Mutat. 2017 Apr;38(4):439-450. doi: 10.1002/humu.23174. Epub 2017 Jan 27.

Transdifferentiation of Human Dermal Fibroblasts to Smooth Muscle-Like Cells to Study the Effect of MYH11 and ACTA2 Mutations in Aortic Aneurysms.

Author information

1
Department of Surgery, Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, The Netherlands.
2
Department of Physiology, Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, The Netherlands.
3
Department of Clinical Genetics, Institute for Cardiovascular Research, VU University Medical Center, Amsterdam, The Netherlands.
4
Department of Oral Cell Biology, ACTA University of Amsterdam and VU University Amsterdam, MOVE Research Institute, Amsterdam, The Netherlands.

Abstract

Mutations in genes encoding proteins of the smooth muscle cell (SMC) contractile apparatus contribute to familial aortic aneurysms. To investigate the pathogenicity of these mutations, SMC are required. We demonstrate a novel method to generate SMC-like cells from human dermal fibroblasts by transdifferentiation to study the effect of variants in genes encoding proteins of the SMC contractile apparatus (ACTA2 and MYH11) in patients with aortic aneurysms. Dermal fibroblasts from seven healthy donors and cells from seven patients with MYH11 or ACTA2 variants were transdifferentiated into SMC-like cells within a 2-week duration using 5 ng/ml TGFβ1 on a scaffold containing collagen and elastin. The induced SMC were comparable to primary human aortic SMC in mRNA expression of SMC markers which was confirmed on the protein level by immunofluorescence quantification analysis and Western blotting. In patients with MYH11 or ACTA2 variants, the effect of intronic variants on splicing was demonstrated on the mRNA level in the induced SMC, allowing classification into pathogenic or nonpathogenic variants. In conclusion, direct conversion of human dermal fibroblasts into SMC-like cells is a highly efficient method to investigate the pathogenicity of variants in proteins of the SMC contractile apparatus.

KEYWORDS:

aortic aneurysms; contractile cytoskeleton; pathogenic variant; smooth muscle cells; transdifferentiation

PMID:
28074631
DOI:
10.1002/humu.23174
[Indexed for MEDLINE]

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