Molecular and genetic dissection of dilated cardiomyopathy
Date of Issue2015
School of Biological Sciences
National Heart Research Institute Singapore
Dilated Cardiomyopathy (DCM), caused by mutations in more than 30 genes, is a life– threatening disease with an estimated prevalence of 1:250. TTN-truncating variants (TTNtv) by nonsense mutations are the most common (~27%) genetic cause of DCM. Around one in 50 people have TTNtv. However, only far end TTNtv are associated with DCM and the mechanism is still unknown. We have identified up-regulation of melusin, integrin β1D and phospho-Akt in the heart samples of TTNtv related DCM patients. In this study, we aim to investigate the interaction of melusin and phospho-Akt to understand how they are involved in DCM. We generated cardiac specific, and actin/GFP co-expressing plasmid to incorporate melusin miRNA cassette. Quantitative real time PCR (Q-PCR) was used to test most potent miRNA cassette and western blot was used to test the effect of melusin inhibition on phospho-Akt level at H9C2 cells. We also tested the effect of overexpression of melusin in H9C2 cells and found that up-regulated melusin activates Akt whereas down-regulation inhibits Akt phosphorylation. This study, for the first time, established the linkage of melusin and phospho-Akt in TTNtv related DCM, which will facilitate our understanding of DCM and provide clues for targeted drug design.
Final Year Project (FYP)