Presentation Information
[II-TKIS-P-4]Examining The Effect Of Laminin On The Differentiation And Behaviour Of In Vitro Cardiomyocytes
○Clara C. Bick1,2, Masaki Shigeta1, Kenta Yashiro1 (1.Anatomy & Developmental Biology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 2.Faculty of Health, Medicine, and Life Science, Maastricht University)
Keywords:
Cardiomyocyte,Differentiation,Laminin
Laminin is a key component of the basement membrane and plays essential roles in cell adhesion, differentiation and mechanotransduction through integrin-mediated interactions. It is also vital for muscle development. Impaired laminin function, as seen in LAMA2-related muscular dystrophy, leads to progressive muscle weakness and atrophy. With advances in respiratory management, cardiac involvement in these patients is reported increasingly.Although LAMA2 has been extensively studied in skeletal and smooth muscle, its role in cardiac muscle remains largely unexplored, despite being a major component of the myocardial extracellular matrix (ECM).This study investigated the role of LAMA2 in mouse embryonic stem cell-derived cardiomyocytes (CMs).A novel Lama2 knockout (KO) model was generated by targeting exon 3 with CRISPR-based methods. KO and wild-type (WT) cells were differentiated into CMs via embryoid body formation using the hanging drop method. Immunostaining confirmed the absence of LAMA2 in KO-derived CMs.Morphological examination showed that KO and WT cells differentiated comparably at early phases. However, with prolonged culture, KO CMs presented reduced maturation and impaired beating activity, consistent with previous studies of other laminin chain KO.This phenotype suggests a gradual decline in CM function and may help explain the progressive cardiac symptoms observed in patients with LAMA2-related muscular dystrophy.RNA-seq data analysis of KO CMs is currently underway to assess the impact of Lama2 loss on gene expression and cardiac signalling pathways.