Molecular and genetic dissection of dilated cardiomyopathy

• Main Author: Hafizah Khalidh
• Other Authors: Li Guang
• Format: Final Year Project
• Language: English
• Published: 2015
• Subjects: DRNTU::Science::Biological sciences::Genetics
• Online Access: http://hdl.handle.net/10356/62952
• Institution: Nanyang Technological University

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.