Targeted Disruption of HLA Genes via CRISPR-Cas9 Generates iPSCs with Enhanced Immune Compatibility

Cell Stem Cell. 2019 Apr 4;24(4):566-578.e7. doi: 10.1016/j.stem.2019.02.005. Epub 2019 Mar 7.

Abstract

Induced pluripotent stem cells (iPSCs) have strong potential in regenerative medicine applications; however, immune rejection caused by HLA mismatching is a concern. B2M gene knockout and HLA-homozygous iPSC stocks can address this issue, but the former approach may induce NK cell activity and fail to present antigens, and it is challenging to recruit rare donors for the latter method. Here, we show two genome-editing strategies for making immunocompatible donor iPSCs. First, we generated HLA pseudo-homozygous iPSCs with allele-specific editing of HLA heterozygous iPSCs. Second, we generated HLA-C-retained iPSCs by disrupting both HLA-A and -B alleles to suppress the NK cell response while maintaining antigen presentation. HLA-C-retained iPSCs could evade T cells and NK cells in vitro and in vivo. We estimated that 12 lines of HLA-C-retained iPSCs combined with HLA-class II knockout are immunologically compatible with >90% of the world's population, greatly facilitating iPSC-based regenerative medicine applications.

Keywords: B2M; CIITA; CRISPR-Cas9; HLA; KIR; NK cell; allogenic; genome editing; iPS cell; immune rejection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics*
  • Cell Line
  • Female
  • Gene Editing*
  • HLA Antigens / genetics*
  • HLA Antigens / immunology
  • Histocompatibility / immunology*
  • Humans
  • Induced Pluripotent Stem Cells / immunology*
  • Induced Pluripotent Stem Cells / metabolism*
  • Male
  • Mice
  • Mice, Inbred NOD

Substances

  • HLA Antigens