Generation of anti-GD2 CAR macrophages from human pluripotent stem cells for cancer immunotherapies
Jue Zhang, Sarah Webster, Bret Duffin, Matthew N. Bernstein, John Steill, Scott Swanson, Matthew H. Forsberg, Jennifer M. Bolin, Matthew E. Brown, Aditi Majumder, Christian M. Capitini, Ron Stewart, James A. Thomson, Igor I. Slukvin
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.