دورية أكاديمية
أعرض في EDS

Multiplexed Functional Assessments of MYH7 Variants in Human Cardiomyocytes.

التفاصيل البيبلوغرافية
العنوان: Multiplexed Functional Assessments of MYH7 Variants in Human Cardiomyocytes.
المؤلفون: Friedman CE; Institute 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.).; 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.; Department of Medicine/Cardiology (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., Fayer S; Department of Genome Sciences (S.F., S.P., L.M.S., D.M.F.), University of Washington, Seattle., Pendyala S; Department of Genome Sciences (S.F., S.P., L.M.S., D.M.F.), University of Washington, Seattle., Chien WM; Institute 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.).; 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.; Department of Medicine/Cardiology (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.; Cardiology/Hospital Specialty Medicine, VA Puget Sound HCS, Seattle, WA (W.-M.C., S.D.F., K.-C.Y.)., Loiben A; Institute 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.).; 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.; Department of Medicine/Cardiology (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., Tran L; Institute 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.).; 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.; Department of Medicine/Cardiology (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., Chao LS; Institute 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.).; 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.; Department of Medicine/Cardiology (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., McKinstry A; Institute 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.).; 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.; Department of Medicine/Cardiology (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., Ahmed D; Institute 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.).; 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.; Department of Medicine/Cardiology (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., Farris SD; 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.; Department of Medicine/Cardiology (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.; Cardiology/Hospital Specialty Medicine, VA Puget Sound HCS, Seattle, WA (W.-M.C., S.D.F., K.-C.Y.)., Stempien-Otero A; 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.; Department of Medicine/Cardiology (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., Jonlin EC; Institute 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.)., Murry CE; Institute 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.).; 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.; Department of Medicine/Cardiology (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.; Department of Laboratory Medicine and Pathology (C.E.M.), University of Washington, Seattle.; Department of Bioengineering (C.E.M., D.M.F.), University of Washington, Seattle., Starita LM; Department of Genome Sciences (S.F., S.P., L.M.S., D.M.F.), University of Washington, Seattle.; Brotman Baty Institute for Precision Medicine, Seattle, WA (L.M.S., D.M.F.)., Fowler DM; Department of Genome Sciences (S.F., S.P., L.M.S., D.M.F.), University of Washington, Seattle.; Department of Bioengineering (C.E.M., D.M.F.), University of Washington, Seattle.; Brotman Baty Institute for Precision Medicine, Seattle, WA (L.M.S., D.M.F.)., Yang KC; Institute 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.).; 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.; Department of Medicine/Cardiology (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.; Cardiology/Hospital Specialty Medicine, VA Puget Sound HCS, Seattle, WA (W.-M.C., S.D.F., K.-C.Y.).
المصدر: Circulation. Genomic and precision medicine [Circ Genom Precis Med] 2024 Apr; Vol. 17 (2), pp. e004377. Date of Electronic Publication: 2024 Feb 16.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Lippincott Williams & Wilkins Country of Publication: United States NLM ID: 101714113 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2574-8300 (Electronic) Linking ISSN: 25748300 NLM ISO Abbreviation: Circ Genom Precis Med Subsets: MEDLINE
أسماء مطبوعة: Original Publication: [Baltimore, MD] : Lippincott Williams & Wilkins, [2018]-
مواضيع طبية MeSH: Induced Pluripotent Stem Cells*/metabolism , Cardiomyopathy, Hypertrophic*/genetics , Cardiomyopathy, Hypertrophic*/metabolism, Humans ; Myocytes, Cardiac/metabolism ; Myosin Heavy Chains/genetics ; Cell Differentiation/genetics ; Cardiac Myosins/genetics
مستخلص: 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.
Competing Interests: Disclosures Dr Murry is an equity holder in Sana Biotechnology and StemCardia. The content is solely the responsibility of the authors and does not necessarily represent the official views of the Department of Veterans Affairs or the US Government.
التعليقات: Comment in: Circ Genom Precis Med. 2024 Apr;17(2):e004599. doi: 10.1161/CIRCGEN.124.004599. (PMID: 38497213)
References: JACC Basic Transl Sci. 2018 Dec 31;3(6):728-740. (PMID: 30623132)
Am J Hum Genet. 2021 Dec 2;108(12):2248-2258. (PMID: 34793697)
Physiol Genomics. 2020 Jul 1;52(7):293-303. (PMID: 32567507)
J Mol Cell Cardiol. 2023 Jun;179:60-71. (PMID: 37019277)
Annu Rev Genet. 2022 Nov 30;56:441-465. (PMID: 36055970)
J Community Genet. 2020 Apr;11(2):139-145. (PMID: 31432391)
Bioinformatics. 2014 Mar 1;30(5):614-20. (PMID: 24142950)
Front Cell Dev Biol. 2022 Jun 16;10:894635. (PMID: 35784482)
Circulation. 2018 Dec 11;138(24):2852-2854. (PMID: 30565988)
Proc Natl Acad Sci U S A. 2006 Nov 21;103(47):17713-7. (PMID: 17095604)
Genet Med. 2023 Aug;25(8):100866. (PMID: 37347242)
Am J Hum Genet. 2017 Sep 7;101(3):315-325. (PMID: 28886340)
Nat Genet. 2018 Jun;50(6):874-882. (PMID: 29785012)
J Mol Evol. 2016 Jan;82(1):11-6. (PMID: 26584803)
Nat Protoc. 2017 Jan;12(1):15-31. (PMID: 27906170)
Proc Natl Acad Sci U S A. 2021 Jun 15;118(24):. (PMID: 34117120)
Nucleic Acids Res. 2015 Jan;43(Database issue):D512-20. (PMID: 25514926)
Genet Med. 2021 Aug;23(8):1381-1390. (PMID: 34012068)
Cell Stem Cell. 2018 Oct 4;23(4):586-598.e8. (PMID: 30290179)
Nature. 2021 Nov;599(7883):91-95. (PMID: 34707284)
Eur Heart J. 2018 Nov 14;39(43):3879-3892. (PMID: 29741611)
Genet Med. 2017 Feb;19(2):192-203. (PMID: 27532257)
Stem Cells. 2020 Feb;38(2):174-186. (PMID: 31664757)
FEBS Lett. 1999 Nov 19;461(3):246-52. (PMID: 10567705)
Circulation. 2022 Apr 19;145(16):1238-1253. (PMID: 35384713)
Nucleic Acids Res. 2016 Jan 4;44(D1):D862-8. (PMID: 26582918)
J Mol Biol. 2005 Oct 21;353(2):459-73. (PMID: 16169011)
Circulation. 2020 Mar 10;141(10):828-842. (PMID: 31983222)
Genet Med. 2018 Mar;20(3):351-359. (PMID: 29300372)
Circulation. 2020 Dec 8;142(23):2262-2275. (PMID: 33025817)
Front Cardiovasc Med. 2022 Feb 21;9:816330. (PMID: 35265683)
Int J Mol Sci. 2023 Mar 03;24(5):. (PMID: 36902340)
Genet Med. 2015 May;17(5):405-24. (PMID: 25741868)
Trends Mol Med. 2016 Nov;22(11):925-934. (PMID: 27742414)
J Mol Cell Cardiol. 2020 Aug;145:43-53. (PMID: 32531470)
Cell. 2008 Feb 22;132(4):661-80. (PMID: 18295582)
Circ Cardiovasc Genet. 2014 Aug;7(4):434-43. (PMID: 25031304)
J Am Coll Cardiol. 2020 Dec 22;76(25):e159-e240. (PMID: 33229116)
J Law Biosci. 2018 Jan 22;4(3):648-657. (PMID: 29868193)
Genet Med. 2015 Nov;17(11):880-8. (PMID: 25611685)
Anat Rec (Hoboken). 2014 Sep;297(9):1670-80. (PMID: 25125180)
معلومات مُعتمدة: RM1 HG010461 United States HG NHGRI NIH HHS; T32 HG000035 United States HG NHGRI NIH HHS; F32 HL164108 United States HL NHLBI NIH HHS; R01 HL148081 United States HL NHLBI NIH HHS; IK2 BX004642 United States BX BLRD VA; T32 GM007266 United States GM NIGMS NIH HHS
فهرسة مساهمة: Keywords: cardiomyopathy, hypertrophic; deep mutational scanning; induced pluripotent stem cells; myosin heavy chain 7
المشرفين على المادة: EC 3.6.4.1 (Myosin Heavy Chains)
0 (MYH7 protein, human)
EC 3.6.1.- (Cardiac Myosins)
تواريخ الأحداث: Date Created: 20240216 Date Completed: 20240418 Latest Revision: 20240626
رمز التحديث: 20240626
مُعرف محوري في PubMed: PMC11196868
DOI: 10.1161/CIRCGEN.123.004377
PMID: 38362799
قاعدة البيانات: MEDLINE
الوصف
تدمد:2574-8300
DOI:10.1161/CIRCGEN.123.004377