Comparison of protocols to generate induced pluripotent stem cell-derived cardiomyocytes and validation of a RNA-seq dataset in in vivo models of arrhythmogenic cardiomyopathy.

Arrhythmogenic cardiomyopathy (ACM) is a rare genetic disease, characterized by ventricular arrhythmias and myocardial fibrofatty replacement. Mechanisms underling its pathogenesis are still unknown, however, useful novel insights can arise integrating human in vitro and murine in vivo models. In this context, reproducibility and comparability of the data are crucial. Human induced pluripotent stem cell (hiPSCs) technology is a valuable source for in vitro models preserving the patient-specific background. In the first part of this thesis, two protocols for differentiation into hiPSC-derived cardiomyocytes were compared, providing information regarding their robustness, and guiding the choice of which to use. The results showed validity for both, with higher beating properties for the protocol previously used by our group than for the protocol used in Goettingen Medical Centre. The second part of this thesis aimed gaining new insights in ACM pathogenesis through an RNA-seq dataset validation from mouse heart samples carrying the p.Q563* DSG2 mutation associated with ACM. Despite only 3 of the 27 analysed genes were validated (Dsg2, Des, Tmem88), differences between three- and six- month-old mice were detected in mitochondria, apoptosis and MAPK pathway related genes, suggesting possible changes underlining pathogenesis temporal evolution, that could be further investigated in the future.