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Multiplexed Functional Assessments of MYH7 Variants in Human Cardiomyocytes
Clayton E Friedman1,2,3, Shawn Fayer4, Sriram Pendyala4
1Institute for Stem Cell and Regenerative Medicine, University of Washington, School of Medicine, Seattle (C.E.F., W.-M.C., A.L., L.T., L.S.C., A.M., D.A., E.C.J., C.E.M., K.-C.Y.).
Circulation. Genomic and Precision Medicine
|February 16, 2024
Summary
This study developed a new method to test MYH7 gene variants using human stem cells, improving the understanding of hypertrophic cardiomyopathy and reducing unknown genetic variants in patients.
Area of Science:
- Cardiovascular Genetics
- Stem Cell Biology
- Molecular Cardiology
Background:
- Pathogenic variants in MYH7 (myosin heavy chain 7) cause hypertrophic cardiomyopathy.
- Most MYH7 missense variants are of unknown significance, hindering clinical diagnosis.
- Previous methods lacked efficiency for deep mutational scanning in human-induced pluripotent stem cells (hiPSCs).
Purpose of the Study:
- To develop a deep mutational scanning method for MYH7 variants using hiPSC-derived cardiomyocytes.
- To assess the functional impact of MYH7 missense variants in a disease-relevant cellular model.
- To reduce the burden of variants of unknown significance for patients and clinicians.
Main Methods:
- Generated an hiPSC library with 113 MYH7 codon variants using CRISPRa.
- Differentiated hiPSCs into cardiomyocytes for functional assessment.
- Assessed beta myosin heavy chain (β-MHC) variant abundance and cardiomyocyte survival via massively parallel sequencing.
Main Results:
- Multiplexed assessment of β-MHC abundance and cardiomyocyte survival accurately distinguished pathogenic from synonymous MYH7 variants.
- Functional data were generated for 4 variants of unknown significance and 58 novel MYH7 missense variants.
- Established β-MHC protein loss in a hypertrophic cardiomyopathy heart with a pathogenic MYH7 variant.
Conclusions:
- Leveraged hiPSC-derived cardiomyocytes for multiplexed assessment of MYH7 missense variants.
- Developed phenotyping strategies enabling deep mutational scanning for clinically actionable genes.
- This approach can decrease the clinical burden of variants of unknown significance.
