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

Kak K Yeung, Natalija Bogunovic, Niels Keekstra, Adriaan A M Beunders, Jorrit Pals, Kim van der Kuij, Eline Overwater, Willem Wisselink, Jan D Blankensteijn, Victor W M van Hinsbergh, Rene J P Musters, Gerard Pals, Dimitra Micha, Behrouz Zandieh-Doulabi

Research output: Contribution to journalArticleAcademicpeer-review

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.

Original languageEnglish
Pages (from-to)439-450
Number of pages12
JournalHuman Mutation
Volume38
Issue number4
DOIs
Publication statusPublished - Apr 2017

Cite this

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title = "Transdifferentiation of Human Dermal Fibroblasts to Smooth Muscle-Like Cells to Study the Effect of MYH11 and ACTA2 Mutations in Aortic Aneurysms",
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 = "Journal Article",
author = "Yeung, {Kak K} and Natalija Bogunovic and Niels Keekstra and Beunders, {Adriaan A M} and Jorrit Pals and {van der Kuij}, Kim and Eline Overwater and Willem Wisselink and Blankensteijn, {Jan D} and {van Hinsbergh}, {Victor W M} and Musters, {Rene J P} and Gerard Pals and Dimitra Micha and Behrouz Zandieh-Doulabi",
note = "{\circledC} 2017 WILEY PERIODICALS, INC.",
year = "2017",
month = "4",
doi = "10.1002/humu.23174",
language = "English",
volume = "38",
pages = "439--450",
journal = "Human Mutation",
issn = "1059-7794",
publisher = "Wiley-Liss Inc.",
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Transdifferentiation of Human Dermal Fibroblasts to Smooth Muscle-Like Cells to Study the Effect of MYH11 and ACTA2 Mutations in Aortic Aneurysms. / Yeung, Kak K; Bogunovic, Natalija; Keekstra, Niels; Beunders, Adriaan A M; Pals, Jorrit; van der Kuij, Kim; Overwater, Eline; Wisselink, Willem; Blankensteijn, Jan D; van Hinsbergh, Victor W M; Musters, Rene J P; Pals, Gerard; Micha, Dimitra; Zandieh-Doulabi, Behrouz.

In: Human Mutation, Vol. 38, No. 4, 04.2017, p. 439-450.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Transdifferentiation of Human Dermal Fibroblasts to Smooth Muscle-Like Cells to Study the Effect of MYH11 and ACTA2 Mutations in Aortic Aneurysms

AU - Yeung, Kak K

AU - Bogunovic, Natalija

AU - Keekstra, Niels

AU - Beunders, Adriaan A M

AU - Pals, Jorrit

AU - van der Kuij, Kim

AU - Overwater, Eline

AU - Wisselink, Willem

AU - Blankensteijn, Jan D

AU - van Hinsbergh, Victor W M

AU - Musters, Rene J P

AU - Pals, Gerard

AU - Micha, Dimitra

AU - Zandieh-Doulabi, Behrouz

N1 - © 2017 WILEY PERIODICALS, INC.

PY - 2017/4

Y1 - 2017/4

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1002/humu.23174

DO - 10.1002/humu.23174

M3 - Article

VL - 38

SP - 439

EP - 450

JO - Human Mutation

JF - Human Mutation

SN - 1059-7794

IS - 4

ER -