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Author ORCID Identifier

https://orcid.org/0000-0001-7882-4921

Date of Award

7-2022

Degree Type

Thesis

Degree Name

Master of Interdisciplinary Studies (MIS)

Department

Biomedical Sciences

First Advisor

Dr. Hongyu Qiu

Second Advisor

Dr. Chunying Li

Abstract

Recent studies have demonstrated that the Valosin-containing protein (VCP), an ATPase – associated protein, plays a protective role in the heart against cardiomyopathies and injuries caused by various cardiac stresses, including ischemia and pressure overload. However, the underlying molecular mechanisms remain largely unknown. It has been shown that VCP presents in stressed hearts. Our hypothesis is that VCP plays an essential role in cardiomyocyte growth and survival through a dual regulatory effect on the signaling of the mechanistic target of rapamycin (mTOR). Thus, we used a functional gain and loss strategy to determine the critical role of VCP in regulating mTOR signaling in cardiomyocytes in vitro. By using recombined adeno-virus system (Ad-CMV-GFP-VCP for overexpression and Ad-U6- siRNA-VCP for knockdown), we successfully overexpressed VCP and knocked down VCP in H9C2 cells respectively. This study aimed to better understand how VCP overexpression and knockdown affected certain proteins involved in the mTOR pathway – in particular phosphorylated Akt (pAkt T308 and pAkt S473), Raptor, Rictor and mTOR. Western blots were also done to confirm VCP knockdown and overexpression with GaPDH as the loading control. This preliminary study, although did not yield substantial results, can be designed as an in vitro system to better understand VCP and mTOR interactions.

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