Go to The Journal of Clinical Investigation
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
  • Current issue
  • Past issues
  • By specialty
    • COVID-19
    • Cardiology
    • Immunology
    • Metabolism
    • Nephrology
    • Oncology
    • Pulmonology
    • All ...
  • Videos
  • Collections
    • Resource and Technical Advances
    • Clinical Medicine
    • Reviews
    • Editorials
    • Perspectives
    • Top read articles
  • JCI This Month
    • Current issue
    • Past issues

  • Current issue
  • Past issues
  • Specialties
  • In-Press Preview
  • Editorials
  • Viewpoint
  • Top read articles
  • About
  • Editors
  • Consulting Editors
  • For authors
  • Publication ethics
  • Transfers
  • Advertising
  • Job board
  • Contact
Base editing repairs an SGCA mutation in human primary muscle stem cells
Helena Escobar, … , Florian Heyd, Simone Spuler
Helena Escobar, … , Florian Heyd, Simone Spuler
Published April 13, 2021
Citation Information: JCI Insight. 2021;6(10):e145994. https://doi.org/10.1172/jci.insight.145994.
View: Text | PDF
Research Article Stem cells Therapeutics

Base editing repairs an SGCA mutation in human primary muscle stem cells

  • Text
  • PDF
Abstract

Skeletal muscle can regenerate from muscle stem cells and their myogenic precursor cell progeny, myoblasts. However, precise gene editing in human muscle stem cells for autologous cell replacement therapies of untreatable genetic muscle diseases has not yet been reported. Loss-of-function mutations in SGCA, encoding α-sarcoglycan, cause limb-girdle muscular dystrophy 2D/R3, an early-onset, severe, and rapidly progressive form of muscular dystrophy affecting both male and female patients. Patients suffer from muscle degeneration and atrophy affecting the limbs, respiratory muscles, and heart. We isolated human muscle stem cells from 2 donors, with the common SGCA c.157G>A mutation affecting the last coding nucleotide of exon 2. We found that c.157G>A is an exonic splicing mutation that induces skipping of 2 coregulated exons. Using adenine base editing, we corrected the mutation in the cells from both donors with > 90% efficiency, thereby rescuing the splicing defect and α-sarcoglycan expression. Base-edited patient cells regenerated muscle and contributed to the Pax7+ satellite cell compartment in vivo in mouse xenografts. Here, we provide the first evidence to our knowledge that autologous gene–repaired human muscle stem cells can be harnessed for cell replacement therapies of muscular dystrophies.

Authors

Helena Escobar, Anne Krause, Sandra Keiper, Janine Kieshauer, Stefanie Müthel, Manuel García de Paredes, Eric Metzler, Ralf Kühn, Florian Heyd, Simone Spuler

×

Figure 6

SGCA c.157Grep patient MuSC regenerate muscle in vivo.

Options: View larger image (or click on image) Download as PowerPoint

SGCA c.157Grep patient MuSC regenerate muscle in vivo.
(A) SGCA c.157Gr...
(A) SGCA c.157Grep patient MuSC were injected into preirradiated anterior tibial muscles of immunocompromised NSG mice. Grafted muscles were collected for analysis after 19 days. (A and B) Grafted muscles were immunostained with antibodies that specifically recognize human Lamin A/C and human Spectrin, labeling donor nuclei and donor-derived myofibers, respectively. In total, 31–87 human nuclei and 33–72 human myofibers were found per section (n = 2). (C) Grafted muscles were immunostained with antibodies against human Lamin A/C and Desmin. (D) Grafted muscles were immunostained with antibodies against human Spectrin and α-sarcoglycan. (E) Grafted muscles were immunostained with antibodies against human Lamin A/C, Pax7, and Laminin to identify satellite cells of human origin located in the stem cell niche under the basal lamina (n = 2). Human Pax7+ satellite cells are indicated by arrows. Nuclei were counterstained with Hoechst. Scale bars: 100 μm (A), 50 μm (B), 20 μm (C and D), 5 μm (E).

Copyright © 2023 American Society for Clinical Investigation
ISSN 2379-3708

Sign up for email alerts