TY - JOUR
T1 - Molecular phenotyping and functional assessment of smooth muscle-like cells with pathogenic variants in aneurysm genes ACTA2, MYH11, SMAD3 and FBN1
AU - Burger, Joyce
AU - Bogunovic, Natalija
AU - De Wagenaar, Nathalie P.
AU - Liu, Hui
AU - Van Vliet, Nicole
AU - Ijpma, Arne
AU - Maugeri, Alessandra
AU - Micha, Dimitra
AU - Verhagen, Hence J.M.
AU - Ten Hagen, Timo L.M.
AU - Majoor-Krakauer, Danielle
AU - Van Der Pluijm, Ingrid
AU - Essers, Jeroen
AU - Yeung, Kak K.
N1 - Funding:
The authors would like to thank Stichting Lijf en Leven (Genexpressie analyse ter detective van de moleculaire mechanismen van aneurysmavorming—GAMMA), Erasmus MC Mrace grant and Amsterdam Cardiovascular Sciences Institute (ICaR AiO 2015 PhD grant) for supporting this research.
Publisher Copyright:
© 2021 The Author(s) 2021.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Aortic aneurysms (AAs) are pathological dilatations of the aorta. Pathogenic variants in genes encoding for proteins of the contractile machinery of vascular smooth muscle cells (VSMCs), genes encoding proteins of the transforming growth factor beta signaling pathway and extracellular matrix (ECM) homeostasis play a role in the weakening of the aortic wall. These variants affect the functioning of VSMC, the predominant cell type in the aorta. Many variants have unknown clinical significance, with unknown consequences on VSMC function and AA development. Our goal was to develop functional assays that show the effects of pathogenic variants in aneurysm-related genes. We used a previously developed fibroblast transdifferentiation protocol to induce VSMC-like cells, which are used for all assays. We compared transdifferentiated VSMC-like cells of patients with a pathogenic variant in genes encoding for components of VSMC contraction (ACTA2, MYH11), transforming growth factor beta (TGFβ) signaling (SMAD3) and a dominant negative (DN) and two haploinsufficient variants in the ECM elastic laminae (FBN1) to those of healthy controls. The transdifferentiation efficiency, structural integrity of the cytoskeleton, TGFβ signaling profile, migration velocity and maximum contraction were measured. Transdifferentiation efficiency was strongly reduced in SMAD3 and FBN1 DN patients. ACTA2 and FBN1 DN cells showed a decrease in SMAD2 phosphorylation. Migration velocity was impaired for ACTA2 and MYH11 cells. ACTA2 cells showed reduced contractility. In conclusion, these assays for showing effects of pathogenic variants may be promising tools to help reclassification of variants of unknown clinical significance in AA-related genes.
AB - Aortic aneurysms (AAs) are pathological dilatations of the aorta. Pathogenic variants in genes encoding for proteins of the contractile machinery of vascular smooth muscle cells (VSMCs), genes encoding proteins of the transforming growth factor beta signaling pathway and extracellular matrix (ECM) homeostasis play a role in the weakening of the aortic wall. These variants affect the functioning of VSMC, the predominant cell type in the aorta. Many variants have unknown clinical significance, with unknown consequences on VSMC function and AA development. Our goal was to develop functional assays that show the effects of pathogenic variants in aneurysm-related genes. We used a previously developed fibroblast transdifferentiation protocol to induce VSMC-like cells, which are used for all assays. We compared transdifferentiated VSMC-like cells of patients with a pathogenic variant in genes encoding for components of VSMC contraction (ACTA2, MYH11), transforming growth factor beta (TGFβ) signaling (SMAD3) and a dominant negative (DN) and two haploinsufficient variants in the ECM elastic laminae (FBN1) to those of healthy controls. The transdifferentiation efficiency, structural integrity of the cytoskeleton, TGFβ signaling profile, migration velocity and maximum contraction were measured. Transdifferentiation efficiency was strongly reduced in SMAD3 and FBN1 DN patients. ACTA2 and FBN1 DN cells showed a decrease in SMAD2 phosphorylation. Migration velocity was impaired for ACTA2 and MYH11 cells. ACTA2 cells showed reduced contractility. In conclusion, these assays for showing effects of pathogenic variants may be promising tools to help reclassification of variants of unknown clinical significance in AA-related genes.
UR - http://www.scopus.com/inward/record.url?scp=85119481190&partnerID=8YFLogxK
U2 - 10.1093/hmg/ddab190
DO - 10.1093/hmg/ddab190
M3 - Article
C2 - 34244757
AN - SCOPUS:85119481190
SN - 0964-6906
VL - 30
SP - 2286
EP - 2299
JO - Human Molecular Genetics
JF - Human Molecular Genetics
IS - 23
ER -