Modulation of mechanosensitive genes during embryonic aortic arch development

• Published in: Developmental dynamics Vol. 254; no. 3; pp. 222 - 239
• Main Authors: Siddiqui, Hummaira Banu, Golcez, Tansu, Çelik, Merve, Sevgin, Börteçine, Çoban, Mervenur, Süder, İlke, Kaya, Özen, Özören, Nesrin, Pekkan, Kerem
• Format: Journal Article
• Language: English
• Published: Hoboken, USA John Wiley & Sons, Inc 01.03.2025; Wiley Subscription Services, Inc
• Subjects: Animals; Aorta; Aorta, Thoracic - embryology; Aorta, Thoracic - metabolism; Aortic arch; Arteries; Cardiovascular system; Chick Embryo; Diameters; Embryogenesis; embryonic aortic arches; Expression vectors; Extracellular matrix; Gelatinase A; Gene expression; Gene Expression Regulation, Developmental - physiology; gene knockdown; Gene Knockdown Techniques; Genes; genetic mapping; Genetic modification; Growth models; Heart diseases; hemodynamics; Immunohistochemistry; in ovo electroporation; Material properties; mechanobiology; Mechanotransduction, Cellular - genetics; Microvasculature; Phenotypes; siRNA; Transforming Growth Factor beta3 - genetics; Transforming Growth Factor beta3 - metabolism; vascular growth model; Western blotting; mechanobiology; genetic mapping; embryonic aortic arches; gene knockdown; hemodynamics; vascular growth model; in ovo electroporation
• ISSN: 1058-8388; 1097-0177; 1097-0177
• DOI: 10.1002/dvdy.728

Background Early embryonic aortic arches (AA) are a dynamic vascular structures that are in the process of shaping into the great arteries of cardiovascular system. Previously, a time‐lapsed mechanosensitive gene expression map was established for AA subject to altered mechanical loads in the avian embryo. To validate this map, we investigated effects on vascular microstructure and material properties following the perturbation of key genes using an in‐house microvascular gene knockdown system. Results All siRNA vectors show a decrease in the expression intensity of desired genes with no significant differences between vectors. In TGFβ3 knockdowns, we found a reduction in expression intensities of TGFβ3 (≤76%) and its downstream targets such as ELN (≤99.6%), Fbn1 (≤60%), COL1 (≤52%) and COL3 (≤86%) and an increase of diameter in the left AA (23%). MMP2 knockdown also reduced expression levels in MMP2 (≤30%) and a 6‐fold increase in its downstream target COL3 with a decrease in stiffness of the AA wall and an increase in the diameter of the AA (55%). These in vivo measurements were confirmed using immunohistochemistry, western blotting and a computational growth model of the vascular extracellular matrix (ECM). Conclusions Localized spatial genetic modification of the aortic arch region governs the vascular phenotype and ECM composition of the embryo and can be integrated with mechanically‐induced congenital heart disease models. Key Findings Among other findings, this article successfully validates the previously presented time resolved gene network map. The study also indicates the role of key genes (MMP2 and TGFb3) involved in development of the aortic arches. The results clearly indicate a direct relation between stiffness and elasticity with the expression levels of MMP2 and TGFb3 respectively. The simulation model of the aortic arch along with the results presented in the current study provides evidence of the validity of the previously presented gene network map and improve our predictive ability for the non‐target genes.