Saturation Mutagenesis of MYH7 to Identify Hypertrophy-causing Variants

Hannah De Jong, Stanford University

Photo of Hannah De Jong

Hypertrophic cardiomyopathy (HCM) is a common inherited cardiac condition, characterized by enlargement of the cells in the left ventricular wall of the heart. Many cases of HCM are caused by genetic variants in the gene MYH7, which encodes beta cardiac myosin. However, hundreds of rare variants are also observed in MYH7, and only a subset cause HCM. A method to predict pathogenic variants would facilitate better clinical diagnosis and improve our understanding of MYH7 biology.

We therefore aim to screen all possible single nucleotide variants (SNVs) in MYH7 using iPSC-cardiomyocytes as a model system. We have created a pool of variant oligonucleotides to systematically generate MYH7 SNVs for expression in iPSC-cardiomyocytes. We will then phenotype the resulting cells for hypertrophy in high-throughput using a microfluidic sorting device developed in the lab. Deep sequencing to identify which MYH7 variants are present in the largest cells will yield candidate variants for HCM pathogenicity.

Abstract Author(s): Hannah De Jong, Shirley Sutton, Euan Ashley