Genomic Safe Harbor Expression of PAX7 for the Generation of Engraftable Myogenic Progenitors

Hyunkee Kim, Sridhar Selvaraj, James Kiley, Karim Azzag, Bayardo I. Garay, Rita C.R. Perlingeiro

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Inducible expression of PAX7 in differentiating pluripotent stem cells (PSCs) allows massively scalable generation of human myogenic progenitors, which upon transplantation into dystrophic muscles give rise to donor-derived myofibers and satellite cells. Therefore, PSC-derived PAX7+ myogenic progenitors represent an attractive therapeutic approach to promote muscle regeneration. Work to date has used lentiviral vectors (LVs) that randomly integrate inducible PAX7 transgenes. Here, we investigated whether equivalent induction of the myogenic program could be achieved by targeting the PAX7 transgene into genomic safe harbor (GSH) sites. Across multiple PSC lines, we find that this approach consistently generates expandable myogenic progenitors in vitro, although scalability of expansion is moderately reduced compared with the LV approach. Importantly, transplantation of GSH-targeted myogenic progenitors produces robust engraftment, comparable with LV counterparts. These findings provide proof of concept for the use of GSH targeting as a potential alternative approach to generate therapeutic PSC-derived myogenic progenitors for clinical applications.

Original languageEnglish (US)
Pages (from-to)10-19
Number of pages10
JournalStem Cell Reports
Volume16
Issue number1
DOIs
StatePublished - Jan 12 2021

Bibliographical note

Funding Information:
We acknowledge the generous support from ADVault 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.). We thank Mark Kotter (University of Cambridge, UK) for targeting and expression plasmids. 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.

Funding Information:
We acknowledge the generous support from ADVault 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.). We thank Mark Kotter (University of Cambridge, UK) for targeting and expression plasmids. 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.

Publisher Copyright:
© 2020 The Authors

Keywords

  • PAX7
  • cell therapy
  • genomic safe harbor sites
  • lentivirus
  • muscle regeneration
  • muscular dystrophies
  • myogenic progenitors
  • pluripotent stem cells
  • transplantation

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

Fingerprint

Dive into the research topics of 'Genomic Safe Harbor Expression of PAX7 for the Generation of Engraftable Myogenic Progenitors'. Together they form a unique fingerprint.

Cite this