Multiplexed Functional Assessments of MYH7 Variants in Human Cardiomyocytes
Open accessAbstract
BACKGROUND: Pathogenic autosomal-dominant missense variants in MYH7 ( myosin heavy chain 7 ), which encodes the sarcomeric protein (β-MHC [beta myosin heavy chain]) expressed in cardiac and skeletal myocytes, are a leading cause of hypertrophic cardiomyopathy and are clinically actionable. However, ≈75% of MYH7 missense variants are of unknown significance. While human-induced pluripotent stem cells (hiPSCs) can be differentiated into cardiomyocytes to enable the interrogation of MYH7 variant effect in a disease-relevant context, deep mutational scanning has not been executed using diploid hiPSC derivates due to low hiPSC gene-editing efficiency. Moreover, multiplexable phenotypes enabling deep mutational scanning of MYH7 variant hiPSC-derived cardiomyocytes are unknown. METHODS: To overcome these obstacles, we used CRISPRa On-Target Editing Retrieval enrichment to generate an hiPSC library containing 113 MYH7 codon variants suitable for deep mutational scanning. We first established that β-MHC protein loss occurs in a hypertrophic cardiomyopathy human heart with a pathogenic MYH7 variant. We then differentiated the MYH7 missense variant hiPSC library to cardiomyocytes for multiplexed assessment of β-MHC variant abundance by massively parallel sequencing and hiPSC-derived cardiomyocyte survival. RESULTS: Both the multiplexed assessment of β-MHC abundance and hiPSC-derived cardiomyocyte survival accurately segregated all known pathogenic variants from synonymous variants. Functional data were generated for 4 variants of unknown significance and 58 additional MYH7 missense variants not yet detected in patients. CONCLUSIONS: This study leveraged hiPSC differentiation into disease-relevant cardiomyocytes to enable multiplexed assessments of MYH7 missense variants for the first time. Phenotyping strategies used here enable the application of deep mutational scanning to clinically actionable genes, which should reduce the burden of variants of unknown significance on patients and clinicians.
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Recent progress has been made towards overcoming problems with iPSC gene editing and promoter silencing.
“although recent progress has been made towards overcoming these problems (87).”
From Massively parallel functional genomic assays in endocrinology: from promise to delivery (2026) · cites this paper as [87]
The Conclusions (page unavailable) describe CRaTER enrichment as enabling the generation of hiPSC variant libraries despite low gene-editing efficiency, supporting progress on the gene-editing problem. The passages do not address promoter silencing or progress toward overcoming it (searched: “promoter silencing” and “silencing promoter”). (LLM, conf=0.84)
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Massively parallel functional genomic assays in endocrinology: from promise to delivery
Xu, J., Aslanzadeh, V., Kudla, G., & Semple, R. K. (2026) · Journal of Molecular Endocrinology · doi:10.1530/jme-25-0134
- Authors
- Friedman, C. E., Fayer, S., Pendyala, S., Chien, W.-M., Loiben, A., Tran, L., Chao, L. S., McKinstry, A., Ahmed, D., Farris, S. D., Stempien-Otero, A., Jonlin, E. C., Murry, C. E., Starita, L. M., Fowler, D. M., & Yang, K.-C.
- Affiliations
- University of Washington (US)
- Center for Cardiovascular Biology (C.E.F., W.-M.C., A.L., L.T., L.S.C., A.M., D.A., S.D.F., A.S.-O., C.E.M., K.-C.Y), University of Washington, Seattle.
- Corresponding author
- Yang, Kai-Chun
- ORCID
- Venue
- Circulation: Genomic and Precision Medicine
- Published
- 2024-04
- Volume
- 17
- Issue
- 2
- Pages
- e004377-e004377
- ISSN
- 2574-8300 (ISSN-L)
- PMC
- PMC11196868
- OpenAlex
- W4391882159
- Cited by
- 17
- References
- 2
- Citations by year
- 2026: 11 · 2025: 5 · 2024: 1
- FWCI
- 4.71
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Licence
- cc-by-nc-nd
- Locations
- Circulation Genomic and Precision Medicine (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- Funders
- U.S. Department of Health and Human Services
- U.S. Department of Veterans Affairs (IK2 BX004642)
- University of Washington
- Seattle Children's Research Institute
- National Institutes of Health
- National Heart, Lung, and Blood Institute (R01HL148081, F32HL164108)
- National Human Genome Research Institute (RM1HG010461, T32HG000035)
- National Institute of General Medical Sciences (T32GM007266)
- National Institute of Diabetes and Digestive and Kidney Diseases (P30DK017047)
- Topics
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiomyopathy and Myosin Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genomics and Rare Diseases
- Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac electrophysiology and arrhythmias
- Keywords
- MYH7, beta-myosin heavy chain, hypertrophic cardiomyopathy, missense mutations, variants of uncertain significance, deep mutational scanning, hiPSC-derived cardiomyocytes, variant effect prediction, cardiomyocyte survival
- MeSH terms
- Cardiomyopathy, Hypertrophic, Cell Differentiation, Humans, Myosin Heavy Chains, Cardiac Myosins, Myocytes, Cardiac, Induced Pluripotent Stem Cells
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