Generation of anti-GD2 CAR macrophages from human pluripotent stem cells for cancer immunotherapies

Stem Cell Reports. 2023 Feb 14;18(2):585-596. doi: 10.1016/j.stemcr.2022.12.012. Epub 2023 Jan 12.

Abstract

Macrophages armed with chimeric antigen receptors (CARs) provide a potent new option for treating solid tumors. However, genetic engineering and scalable production of somatic macrophages remains significant challenges. Here, we used CRISPR-Cas9 gene editing methods to integrate an anti-GD2 CAR into the AAVS1 locus of human pluripotent stem cells (hPSCs). We then established a serum- and feeder-free differentiation protocol for generating CAR macrophages (CAR-Ms) through arterial endothelial-to-hematopoietic transition (EHT). CAR-M produced by this method displayed a potent cytotoxic activity against GD2-expressing neuroblastoma and melanoma in vitro and neuroblastoma in vivo. This study provides a new platform for the efficient generation of off-the-shelf CAR-Ms for antitumor immunotherapy.

Keywords: CAR; GD2; PSCs; chimeric antigen receptor; hemogenic endothelium; immunotherapy; macrophages; melanoma; neuroblastoma; pluripotent stem cells.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Humans
  • Immunotherapy / methods
  • Macrophages / pathology
  • Melanoma* / therapy
  • Neuroblastoma* / pathology
  • Neuroblastoma* / therapy
  • Pluripotent Stem Cells* / pathology
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Chimeric Antigen* / genetics

Substances

  • Receptors, Chimeric Antigen
  • Receptors, Antigen, T-Cell