Vanessa Rodgers
Research Mentor: Adam Helms
Mentor Department: Internal Medicine / Cardiovascular Medicine, Medicine
Author(s): Vanessa Rodgers, Eric Smith, Adam Helms
Session: Session 5 (2:00 PM – 2:50 PM)
Presentation Type: Poster 118
Abstract
Background: Desmoplakin (DSP) cardiomyopathy is a genetic heart disease characterized by arrhythmias and excessive cardiac fibrosis, caused by truncating variants in the DSP gene that reduce DSP protein at cardiomyocyte junctions. While prior in vitro studies have demonstrated that increasing DSP expression can restore cellular phenotypes, these effects have not yet been evaluated in an in vivo mouse model. Objective: The objective of this study is to develop an adeno-associated virus (AAV)-based CRISPR activation (CRISPRa) strategy to restore DSP expression for subsequent in vivo evaluation in mouse models. Methods: Multiple AAV-compatible CRISPRa constructs targeting DSP were designed and assembled using standard cloning techniques, including Gibson and Golden Gate cloning, then sequence-verified prior to viral packaging. Constructs were initially tested in cultured cells to confirm expression and functionality before downstream in vivo application. Anticipated Results: We anticipate that AAV-mediated CRISPRa will increase DSP expression in cardiac tissue, resulting in reduced fibrosis, decreased incidence of arrhythmias, and overall preservation of cardiac structure and function. Conclusions: This work establishes an experimental framework for translating CRISPRa-based DSP restoration strategies from in vitro systems to in vivo models and may inform the development of potential gene therapies for DSP cardiomyopathy.
