@article{7f4e3929ce4d407ba2384f2ea2430aeb,
title = "Base editing correction of hypertrophic cardiomyopathy in human cardiomyocytes and humanized mice",
abstract = "The most common form of genetic heart disease is hypertrophic cardiomyopathy (HCM), which is caused by variants in cardiac sarcomeric genes and leads to abnormal heart muscle thickening. Complications of HCM include heart failure, arrhythmia and sudden cardiac death. The dominant-negative c.1208G>A (p.R403Q) pathogenic variant (PV) in β-myosin (MYH7) is a common and well-studied PV that leads to increased cardiac contractility and HCM onset. In this study we identify an adenine base editor and single-guide RNA system that can efficiently correct this human PV with minimal bystander editing and off-target editing at selected sites. We show that delivery of base editing components rescues pathological manifestations of HCM in induced pluripotent stem cell cardiomyocytes derived from patients with HCM and in a humanized mouse model of HCM. Our findings demonstrate the potential of base editing to treat inherited cardiac diseases and prompt the further development of adenine base editor-based therapies to correct monogenic variants causing cardiac disease.",
author = "Chai, {Andreas C.} and Miao Cui and Francesco Chemello and Hui Li and Kenian Chen and Wei Tan and Ayhan Atmanli and McAnally, {John R.} and Yu Zhang and Lin Xu and Ning Liu and Rhonda Bassel-Duby and Olson, {Eric N.}",
note = "Funding Information: Peripheral blood mononuclear cells from two patients with the MYH7 c.1208G>A (p.R403Q) PV were reprogrammed to iPSCs (HCM1 and HCM2) using Sendai virus. The HCM1 line was derived from a 56-year-old woman with extensive family history of HCM and nonobstructive HCM with a history of reduced left ventricular EF and low maximal oxygen uptake (VO max). A biventricular pacemaker was placed for a complete heart block. The HCM2 line was derived from a 32-year-old man with a history of HCM, an implantable cardioverter-defibrillator and a strong family history of HCM. He has a dilated left atrium but has improved VO max, metabolic equivalent and no evidence of atrial fibrillation by cardiopulmonary exercise testing. These two human iPSC lines were obtained from Joseph C. Wu, MD, PhD at the Stanford Cardiovascular Institute funded by National Institutes of Health grant R24 HL117756. Peripheral blood mononuclear cells from a male HD were reprogrammed to iPSCs at the UT Southwestern Wellstone Myoediting Core using Sendai virus (CytoTune 2.0 Sendai Reprogramming Kit, Thermo Fisher Scientific). To generate isogenic iPSCs containing the MYH7 c.1208G>A (p.R403Q) variant via homology-directed repair, HD iPSCs were nucleofected using the P3 Primary Cell 4D-Nucleofector X Kit (Lonza) with a single-stranded oligodeoxynucleotide template (Integrated DNA Technologies, IDT) including the PV, and the PX458 plasmid encoding SpCas9-P2a-EGFP and a sgRNA targeting MYH7. For base editing correction of HCM1 and HCM2 patient-derived lines, iPSCs were nucleofected with a single plasmid encoding ABEmax-VRQR-P2a-EGFP and h403_sgRNA. After 48 h, GFP + iPSCs were collected by fluorescence-activated cell sorting, clonally expanded, and genotyped by Sanger sequencing. 2 2 Funding Information: We thank the members of the E.N.O. laboratory for helpful discussions; J. C. Wu and the Stanford Cardiovascular Institute for providing the HCM patient-derived iPSCs; C. Seidman and H. Wakimoto for suggestions on intrathoracic AAV9 delivery; T. Lanigan, H. Kopera and R. Agate from the University of Michigan Vector Core for rAAV9 production; C. Llamas and P. Mishra from the Children{\textquoteright}s Medical Center Research Institute for help with Seahorse assays; J. Cabrera and S. Vargas for graphics; D. Martin from Envigo for custom chow consultation; J. Xu and Y. J. Kim from the Children{\textquoteright}s Medical Center Research Institute for performing the Illumina NextSeq sequencing; C. Rodriguez-Caycedo for assistance with iPSCs; the UT Southwestern McDermott Center Sanger Sequencing Core; the UT Southwestern McDermott Center Next-Generation Sequencing Core; the UT Southwestern Flow Cytometry Core; and J. Shelton from the Molecular Histopathology Core for help with histology. This work was supported by grants from the National Institutes of Health (R01HL130253, P50HD087351 and R01HL157281 to E.N.O. and R.B.-D.; F30HL163915 to A.C.C.), the American Heart Association (907611 to A.C.C.), the Foundation Leducq Transatlantic Networks of Excellence in Cardiovascular Research and the Robert A. Welch Foundation (grant 1-0025 to E.N.O.). The E.N.O. laboratory is supported by CureHeart, the British Heart Foundation{\textquoteright}s Big Beat Challenge Award (BBC/F/21/220106). Funding Information: We thank the members of the E.N.O. laboratory for helpful discussions; J. C. Wu and the Stanford Cardiovascular Institute for providing the HCM patient-derived iPSCs; C. Seidman and H. Wakimoto for suggestions on intrathoracic AAV9 delivery; T. Lanigan, H. Kopera and R. Agate from the University of Michigan Vector Core for rAAV9 production; C. Llamas and P. Mishra from the Children{\textquoteright}s Medical Center Research Institute for help with Seahorse assays; J. Cabrera and S. Vargas for graphics; D. Martin from Envigo for custom chow consultation; J. Xu and Y. J. Kim from the Children{\textquoteright}s Medical Center Research Institute for performing the Illumina NextSeq sequencing; C. Rodriguez-Caycedo for assistance with iPSCs; the UT Southwestern McDermott Center Sanger Sequencing Core; the UT Southwestern McDermott Center Next-Generation Sequencing Core; the UT Southwestern Flow Cytometry Core; and J. Shelton from the Molecular Histopathology Core for help with histology. This work was supported by grants from the National Institutes of Health (R01HL130253, P50HD087351 and R01HL157281 to E.N.O. and R.B.-D.; F30HL163915 to A.C.C.), the American Heart Association (907611 to A.C.C.), the Foundation Leducq Transatlantic Networks of Excellence in Cardiovascular Research and the Robert A. Welch Foundation (grant 1-0025 to E.N.O.). The E.N.O. laboratory is supported by CureHeart, the British Heart Foundation{\textquoteright}s Big Beat Challenge Award (BBC/F/21/220106). Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive licence to Springer Nature America, Inc.",
year = "2023",
month = feb,
doi = "10.1038/s41591-022-02176-5",
language = "English (US)",
volume = "29",
pages = "401--411",
journal = "Nature Medicine",
issn = "1078-8956",
publisher = "Nature Publishing Group",
number = "2",
}