CoChat
Sign Up
Cited by 203

The DOI matches its Crossref record; 11 of the 11 references it cites were found in a registry or an index.

Efficient in vivo genome editing prevents hypertrophic cardiomyopathy in mice

Reichart, Daniel, Newby, Gregory A., Wakimoto, Hiroko, Lun, Mingyue, Gorham, Joshua M., Curran, Justin J., Raguram, Aditya, DeLaughter, Daniel M., Conner, David A., Marsiglia, Júlia D. C., Kohli, Sajeev, Chmatal, Lukas, Page, David C., Zabaleta, Nerea, Vandenberghe, Luk, Liu, David R., Seidman, Jonathan G., Seidman, Christine·2023·Nature Medicine
DOI 10.1038/s41591-022-02190-7PMC PMC9941048
Open this paper in CoChat — free account

Abstract

Dominant missense pathogenic variants in cardiac myosin heavy chain cause hypertrophic cardiomyopathy (HCM), a currently incurable disorder that increases risk for stroke, heart failure and sudden cardiac death. In this study, we assessed two different genetic therapies—an adenine base editor (ABE8e) and a potent Cas9 nuclease delivered by AAV9—to prevent disease in mice carrying the heterozygous HCM pathogenic variant myosin R403Q. One dose of dual-AAV9 vectors, each carrying one half of RNA-guided ABE8e, corrected the pathogenic variant in ≥70% of ventricular cardiomyocytes and maintained durable, normal cardiac structure and function. An additional dose provided more editing in the atria but also increased bystander editing. AAV9 delivery of RNA-guided Cas9 nuclease effectively inactivated the pathogenic allele, albeit with dose-dependent toxicities, necessitating a narrow therapeutic window to maintain health. These preclinical studies demonstrate considerable potential for single-dose genetic therapies to correct or silence pathogenic variants and prevent the development of HCM.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/s41591-022-02190-7

Cites

11 of 11 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Show all 11 references

Research works printed but not linked: 44

  1. Garfinkel, A. C., Seidman, J. G. & Seidman, C. E. Genetic pathogenesis of hypertrophic and dilated cardiomyopathy. Heart Fail Clin. 14, 139–146 (2018). · DOI
  2. Walsh, R. et al. Reassessment of Mendelian gene pathogenicity using 7,855 cardiomyopathy cases and 60,706 reference samples. Genet. Med. 19, 192–203 (2017). · DOI
  3. Ho, C. Y. et al. Genotype and lifetime burden of disease in hypertrophic cardiomyopathy: insights from the Sarcomeric Human Cardiomyopathy Registry (SHaRe). Circulation 138, 1387–1398 (2018). · DOI · Paper page
  4. Ho, C. Y. et al. Assessment of diastolic function with Doppler tissue imaging to predict genotype in preclinical hypertrophic cardiomyopathy. Circulation 105, 2992–2997 (2002). · DOI
  5. Michels, M. et al. Diastolic abnormalities as the first feature of hypertrophic cardiomyopathy in Dutch myosin-binding protein C founder mutations. JACC Cardiovasc. Imaging 2, 58–64 (2009). · DOI
  6. Paunovska, K., Loughrey, D. & Dahlman, J. E. Drug delivery systems for RNA therapeutics. Nat. Rev. Genet. 23, 265–280 (2022). · DOI · Paper page
  7. Raguram, A., Banskota, S. & Liu, D. R. Therapeutic in vivo delivery of gene editing agents. Cell 185, 2806–2827 (2022). · DOI
  8. Geisterfer-Lowrance, A. A. et al. A mouse model of familial hypertrophic cardiomyopathy. Science 272, 731–734 (1996). · DOI
  9. Teekakirikul, P. et al. Cardiac fibrosis in mice with hypertrophic cardiomyopathy is mediated by non-myocyte proliferation and requires Tgf-β. J. Clin. Invest. 120, 3520–3529 (2010). · DOI
  10. Mar, J. H., Antin, P. B., Cooper, T. A. & Ordahl, C. P. Analysis of the upstream regions governing expression of the chicken cardiac troponin T gene in embryonic cardiac and skeletal muscle cells. J. Cell Biol. 107, 573–585 (1988). · DOI
  11. Jiang, J., Wakimoto, H., Seidman, J. G. & Seidman, C. E. Allele-specific silencing of mutant Myh6 allele in mice suppresses hypertrophic cardiomyopathy. Science 342, 111–114 (2013). · DOI
  12. Koblan, L. W. et al. In vivo base editing rescues Hutchinson–Gilford progeria syndrome in mice. Nature 589, 608–614 (2021). · DOI
  13. Varga, R. et al. Progressive vascular smooth muscle cell defects in a mouse model of Hutchinson–Gilford progeria syndrome. Proc. Natl Acad. Sci. USA 103, 3250–3255 (2006). · DOI
  14. Levy, J. M. et al. Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses. Nat. Biomed. Eng. 4, 97–110 (2020). · DOI
  15. Miller, S. M. et al. Continuous evolution of SpCas9 variants compatible with non-G PAMs. Nat. Biotechnol. 38, 471–481 (2020). · DOI
  16. Green, E. M. et al. A small-molecule inhibitor of sarcomere contractility suppresses hypertrophic cardiomyopathy in mice. Science 351, 617–621 (2016). · DOI
  17. Sedaghat-Hamedani, F. et al. Clinical outcomes associated with sarcomere mutations in hypertrophic cardiomyopathy: a meta-analysis on 7675 individuals. Clin. Res. Cardiol. 107, 30–41 (2018). · DOI
  18. Wakimoto, H., Seidman, J. G., Foo, R. S. & Jiang, J. AAV9 delivery of shRNA to the mouse heart. Curr. Protoc. Mol. Biol. 115, 23.16.1–23.16.9 (2016). · DOI
  19. Axelsson Raja, A. et al. Ablation of lysophosphatidic acid receptor 1 attenuates hypertrophic cardiomyopathy in a mouse model. Proc. Natl Acad. Sci. USA 119, e2204174119 (2022). · DOI
  20. Raphael, C. E. et al. Cardiovascular magnetic resonance predictors of heart failure in hypertrophic cardiomyopathy: the role of myocardial replacement fibrosis and the microcirculation. J. Cardiovasc. Magn. Reson. 23, 26 (2021). · DOI
  21. Vullaganti, S. et al. Fibrosis in hypertrophic cardiomyopathy patients with and without sarcomere gene mutations. Heart Lung Circ. 30, 1496–1501 (2021). · DOI
  22. Zincarelli, C., Soltys, S., Rengo, G. & Rabinowitz, J. E. Analysis of AAV serotypes 1–9 mediated gene expression and tropism in mice after systemic injection. Mol. Ther. 16, 1073–1080 (2008). · DOI · Paper page
  23. Roşca, M. et al. Right ventricular remodeling, its correlates, and its clinical impact in hypertrophic cardiomyopathy. J. Am. Soc. Echocardiogr. 28, 1329–1338 (2015). · DOI
  24. Nedios, S. et al. Characteristics of left atrial remodeling in patients with atrial fibrillation and hypertrophic cardiomyopathy in comparison to patients without hypertrophy. Sci. Rep. 11, 12411 (2021). · DOI
  25. Tai, D. S., Hu, C., Kim, E. H. & Lipshutz, G. S. Augmentation of transgene-encoded protein after neonatal injection of adeno-associated virus improves hepatic copy number without immune responses. Pediatr. Res. 78, 239–246 (2015). · DOI
  26. Christodoulou, D. C. et al. 5′RNA-Seq identifies Fhl1 as a genetic modifier in cardiomyopathy. J. Clin. Invest. 124, 1364–1370 (2014). · DOI
  27. Lazzarotto, C. R. et al. Defining CRISPR–Cas9 genome-wide nuclease activities with CIRCLE-seq. Nat. Protoc. 13, 2615–2642 (2018). · DOI
  28. Davis, J. R. et al. Efficient in vivo base editing via single adeno-associated viruses with size-optimized genomes encoding compact adenine base editors. Nat. Biomed. Eng. 6, 1272–1283 (2022). · DOI
  29. Jones, W. K. et al. Ablation of the murine alpha myosin heavy chain gene leads to dosage effects and functional deficits in the heart. J. Clin. Invest. 98, 1906–1917 (1996). · DOI
  30. Fatkin, D. et al. Neonatal cardiomyopathy in mice homozygous for the Arg403Gln mutation in the α cardiac myosin heavy chain gene. J. Clin. Invest. 103, 147–153 (1999). · DOI
  31. Neill, W. A., Sewell, D. H., Gosalia, K. P., Kingsley, E. M. & Oxendine, J. M. Atrial coronary hyperemia and ischemia in response to variations in atrial pressure in dogs. Am. Heart J. 106, 554–558 (1983). · DOI
  32. van Bragt, K. A. et al. Dynamic regulation of atrial coronary blood flow in healthy adult pigs. Heart Rhythm 12, 991–1000 (2015). · DOI
  33. Gudmundsson, S. et al. Variant interpretation using population databases: lessons from gnomAD. Hum. Mutat. 43, 1012–1030 (2022). · DOI
  34. Rees, H. A., Wilson, C., Doman, J. L. & Liu, D. R. Analysis and minimization of cellular RNA editing by DNA adenine base editors. Sci. Adv. 5, eaax5717 (2019). · DOI
  35. Gaudelli, N. M. et al. Directed evolution of adenine base editors with increased activity and therapeutic application. Nat. Biotechnol. 38, 892–900 (2020). · DOI
  36. Anzalone, A. V. et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature 576, 149–157 (2019). · DOI · Paper page
  37. Kim, D. Y., Moon, S. B., Ko, J.-H., Kim, Y.-S. & Kim, D. Unbiased investigation of specificities of prime editing systems in human cells. Nucleic Acids Res. 48, 10576–10589 (2020). · DOI
  38. Ho, C. Y. et al. Evolution of hypertrophic cardiomyopathy in sarcomere mutation carriers. Heart 102, 1805–1812 (2016). · DOI
  39. Lock, M. et al. Rapid, simple, and versatile manufacturing of recombinant adeno-associated viral vectors at scale. Hum. Gene Ther. 21, 1259–1271 (2010). · DOI
  40. Sanmiguel, J., Gao, G. & Vandenberghe, L. H. Quantitative and digital droplet-based AAV genome titration. Methods Mol. Biol. 1950, 51–83 (2019). · DOI
  41. Clement, K. et al. CRISPResso2 provides accurate and rapid genome editing sequence analysis. Nat. Biotechnol. 37, 224–226 (2019). · DOI
  42. Korsunsky, I. et al. Fast, sensitive and accurate integration of single-cell data with Harmony. Nat. Methods 16, 1289–1296 (2019). · DOI · Paper page
  43. Bernstein, N. J. et al. Solo: doublet identification in single-cell RNA-seq via semi-supervised deep learning. Cell Syst. 11, 95–101.e5 (2020). · DOI · Paper page
  44. Diroma, M. A., Ciaccia, L., Pesole, G. & Picardi, E. Elucidating the editome: bioinformatics approaches for RNA editing detection. Brief. Bioinform. 20, 436–447 (2019). · DOI

Cited by

Sorted by citations

See all 203

Metadata

Authors
Reichart, D., Newby, G. A., Wakimoto, H., Lun, M., Gorham, J. M., Curran, J. J., Raguram, A., DeLaughter, D. M., Conner, D. A., Marsiglia, J. D. C., Kohli, S., Chmatal, L., Page, D. C., Zabaleta, N., Vandenberghe, L., Liu, D. R., Seidman, J. G., & Seidman, C.
Venue
Nature Medicine
Publisher
Springer Science and Business Media LLC
Published
2023-02
Volume
29
Issue
2
Pages
412-421
ISSN
  • 1078-8956
  • 1546-170X
DOI
10.1038/s41591-022-02190-7
PMC
PMC9941048
Cited by
203
References
11
Language
en
Licence
Funders
  • Howard Hughes Medical Institute
  • U.S. Department of Health & Human Services | NIH | Office of Extramural Research, National Institutes of Health (HL080494, HL084553, U01 AI142756, RM1 HG009490, R35 GM118062)
  • Deutsche Forschungsgemeinschaft (N/A)
  • Helen Hay Whitney Foundation
  • National Science Foundation Graduate Research Award

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review