Grant: Master's Award: Canada Graduate Scholarships

Hypertrophic cardiomyopathy (HCM), defined as the thickening of the heart’s walls, is a cardiovascular disease that affects 1/500 people. Pathogenic HCM is often caused by monogenic heterogenous point mutations in sarcomeric proteins, leading to reduced contractile efficiency of cardiac tissue. Because of the genetic nature of HCM, CRISPR:Cas9 gene editing therapy has been heavily considered as a non-invasive treatment option, however, lacks an efficient delivery system that could effectively target diseased cardiac tissue. Adeno-associated vectors are commonly used for in-vivo studies, however, are viral in nature and are limited by their small carrying capacity for cargo. Microbubbles (MBs) and ultrasound (US) may be a viable alternative for in-vivo delivery of Cas9 for gene therapy. MBs are small gas-filled lipid-encapsulated bubbles that under US conditions, oscillate about their resting size. MBs are currently approved as contrast agents however under the correct US parameters, can be utilized to reversibly permeate cellular membranes. This can allow for nearby otherwise impermeable macromolecules such as proteins to pass through treated tissue. Our aim is to utilize a focused US system and MBs to deliver a Cas9 ribonucleopeptide complex (RNP) to hypertrophic human cardiomyocytes. We will first generate a pathogenic hypertrophic model in human induced pluripotent stem cells (hiPSC).We will then differentiate the pathogenic hiPSC line into diseased cardiomyocytes for treatment using the Cas9:prime editing guide RNA system. MBs and US could revolutionize the field of gene editing therapy by allowing for targeted image-guided therapy of cardiovascular diseases.