Brianna Ferguson
Research Mentor: Adam Helms
Mentor Department: Internal Medicine / Cardiovascular Medicine, Medicine
Author(s): Brianna Ferguson, Eric Smith, Ahmed Elmansi, Adam Helms
Session: Session 2 (10:00 AM – 10:50 AM)
Presentation Type:
Abstract
Desmoplakin (DSP) cardiomyopathy is a heart disease characterized by excessive cardiac fibrosis and arrhythmias. Cardiac fibrosis is the build up of connective tissue within the heart as a result of injury – resulting in heart muscle dysfunction and abnormal electrical conduction. Arrhythmias, a consequence of altered electrical conduction, are irregular beats of the heart that can cause chest pain, shortness of breath and left untreated can result in death. DSP cardiomyopathy is caused by truncating variants in desmoplakin (DSPtv) which result in decreased DSP protein at myocardial cell junctions. Cardiomyocyte muscle bundles harboring DSPtv show increased susceptibility to injury with mechanical stress, such as exercise, and is the hypothezied mechanism of cardiac fibrosis in vivo. The therapies in deveopment to mitigate the progression of DSP cardiomyopathy primarily focus at restoration of DSP protein abundance, for example, gene therapy. The current standard of care focuses on modulating the secondary effects of fibrosis, treating arrhythmias and in extreme cases replacing the entire heart by transplant. To test the effectiveness of therapies that may prevent disease progression, the quantity of DSP within the heart tissue needs to be quantified. In past research methodologies, such as western blotting, quantifying DSP is time and resource intensive, in part due to its low abundance in cardiac tissue. Recently, a new protein assay technique was developed, called AlphaLISA. An AlphaLISA assay utilizes the fluorescent properties of two different types of bioengineered beads, acceptor beads and donor beads, that emit light when in close proximity to each other. By conjugating these beads to antibodies that target a protein of interest, in this case, DSP, the beads are brought together and emit a fluorescent signal. At this time there is not a validated AlphaLISA assay for reproducible measurement of DSP abundance. For my Research Scholars project, I developed and tested combinations of beads, antibodies, and various experimental procedures to create a reliable AlphaLISA assay for DSP that can be used for all future experiments. This tool will become indispensable for researching therapies for desmoplakin cardiomyopathy and to save lives.
