Gene Correction of LGMD2A Patient-Specific iPSCs for the Development of Targeted Autologous Cell Therapy

Sridhar Selvaraj, Neha R. Dhoke, James Kiley, Alba Judith Mateos-Aierdi, Sudheer Tungtur, Ricardo Mondragon-Gonzalez, Grace Killeen, Vanessa K.P. Oliveira, Adolfo López de Munain, Rita C.R. Perlingeiro

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Limb girdle muscular dystrophy type 2A (LGMD2A), caused by mutations in the Calpain 3 (CAPN3) gene, is an incurable autosomal recessive disorder that results in muscle wasting and loss of ambulation. To test the feasibility of an autologous induced pluripotent stem cell (iPSC)-based therapy for LGMD2A, here we applied CRISPR-Cas9-mediated genome editing to iPSCs from three LGMD2A patients to enable correction of mutations in the CAPN3 gene. Using a gene knockin approach, we genome edited iPSCs carrying three different CAPN3 mutations, and we demonstrated the rescue of CAPN3 protein in myotube derivatives in vitro. Transplantation of gene-corrected LGMD2A myogenic progenitors in a novel mouse model combining immunodeficiency and a lack of CAPN3 resulted in muscle engraftment and rescue of the CAPN3 mRNA. Thus, we provide here proof of concept for the integration of genome editing and iPSC technologies to develop a novel autologous cell therapy for LGMD2A.

Original languageEnglish (US)
Pages (from-to)2147-2157
Number of pages11
JournalMolecular Therapy
Volume27
Issue number12
DOIs
StatePublished - Dec 4 2019

Bibliographical note

Funding Information:
We thank the generous support from ADVault, Inc. and MyDirectives.com (R.C.R.P.). This project was also supported by funds from the NIH , grants R01 AR055299 and AR071439 (R.C.R.P.), the Coalition to Cure Calpain 3 (R.C.R.P.), and Instituto Carlos III, Fondo Investigaciones Sanitarias-ALM, PI17/01841 (A.J.M.-A and A.L.d.M). The cytogenetic analyses were performed in the Cytogenomics Shared Resource at the University of Minnesota with support from the comprehensive Masonic Cancer Center NIH grant P30 CA077598-09 . The monoclonal antibody to MHC was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. We thank Melissa Spencer (UCLA) for the C3KO mice and Michael Kyba for critical reading of the manuscript.

Funding Information:
We thank the generous support from ADVault, Inc. and MyDirectives.com (R.C.R.P.). This project was also supported by funds from the NIH, grants R01 AR055299 and AR071439 (R.C.R.P.), the Coalition to Cure Calpain 3 (R.C.R.P.), and Instituto Carlos III, Fondo Investigaciones Sanitarias-ALM, PI17/01841 (A.J.M.-A and A.L.d.M). The cytogenetic analyses were performed in the Cytogenomics Shared Resource at the University of Minnesota with support from the comprehensive Masonic Cancer Center NIH grant P30 CA077598-09. The monoclonal antibody to MHC was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa. We thank Melissa Spencer (UCLA) for the C3KO mice and Michael Kyba for critical reading of the manuscript.

Publisher Copyright:
© 2019 The American Society of Gene and Cell Therapy

Keywords

  • CAPN3
  • Calpain 3
  • autologous cell therapy
  • gene correction
  • genome editing
  • iPSCs
  • induced pluripotent stem cells
  • limb girdle muscular dystrophy type 2A
  • myogenic progenitors

Fingerprint

Dive into the research topics of 'Gene Correction of LGMD2A Patient-Specific iPSCs for the Development of Targeted Autologous Cell Therapy'. Together they form a unique fingerprint.

Cite this