CAR-NK cells derived from cord blood originate mainly from CD56-CD7+CD34-HLA-DR-Lin- NK progenitor cells

Tansri Wibowo; Yosuke Kogue; Shunya Ikeda; Moto Yaga; Mana Tachikawa; Makiko Suga; Shuhei Kida; Kumi Shibata; Kazuhito Tsutsumi; Hiraku Murakami; Yasutaka Ueda; Hisashi Kato; Kentaro Fukushima; Jiro Fujita; Tomoaki Ueda; Shinsuke Kusakabe; Akihisa Hino; Michiko Ichii; Chihaya Imai; Daisuke Okuzaki; Atsushi Kumanogoh; Naoki Hosen

doi:10.1016/j.omtm.2024.101374

CAR-NK cells derived from cord blood originate mainly from CD56^-CD7⁺CD34^-HLA-DR^-Lin^- NK progenitor cells

Mol Ther Methods Clin Dev. 2024 Nov 12;32(4):101374. doi: 10.1016/j.omtm.2024.101374. eCollection 2024 Dec 12.

Authors

Tansri Wibowo¹, Yosuke Kogue^{2

3}, Shunya Ikeda⁴, Moto Yaga¹, Mana Tachikawa⁵, Makiko Suga³, Shuhei Kida³, Kumi Shibata³, Kazuhito Tsutsumi³, Hiraku Murakami³, Yasutaka Ueda³, Hisashi Kato³, Kentaro Fukushima³, Jiro Fujita³, Tomoaki Ueda³, Shinsuke Kusakabe³, Akihisa Hino³, Michiko Ichii³, Chihaya Imai⁶, Daisuke Okuzaki⁷, Atsushi Kumanogoh^{1

4

8

9}, Naoki Hosen^{3

4

8}

Affiliations

¹ Department of Respiratory Medicine and Clinical Immunology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
² Osaka Research Center for Drug Discovery, Otsuka Pharmaceutical Company, Ltd., Osaka 562-0029, Japan.
³ Department of Hematology and Oncology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
⁴ World Premier International Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
⁵ Department of Clinical Laboratory and Biomedical Sciences, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
⁶ Department of Pediatrics, Faculty of Medicine, University of Toyama, Toyama 930-0194, Japan.
⁷ Genome Information Research Center, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan.
⁸ Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Osaka 565-0871, Japan.
⁹ Center for Infectious Disease Education and Research, Osaka University, Osaka 565-0871, Japan.

Abstract

Cord blood (CB)-derived chimeric antigen receptor (CAR)-natural killer (NK) cells targeting CD19 have been shown to be effective against B cell malignancies. While human CD56⁺ NK cells can be expanded in vitro, NK cells can also be differentiated from hematopoietic progenitor cells. It is still unclear whether CAR-NK cells originate from mature NK cells or NK progenitor cells in CB. Here, we determined that CAR-NK cells were predominantly derived from CD56^- NK progenitor cells. We first found that substantial numbers of CD19 CAR-NK cells were produced from CD56^- CB mononuclear cells after in vitro culture for 2 weeks. Single-cell RNA sequencing analysis of CD56^-CD3^-CD14^-CD19^- CB mononuclear cells revealed that these cells could be subdivided into three subpopulations based on the expression of CD34 and human leukocyte antigen (HLA)-DR. NK cells originated primarily from CD34^-HLA-DR^- cells. In addition, among the CD34^-HLA-DR^- cells, only CD7⁺ cells could differentiate into NK cells. These results indicate that CD56^-CD7⁺CD34^-HLA-DR^- lineage marker (Lin)^- cells are the major origin of human CB-derived CAR-NK cells, indicating the importance of developing methods to enhance the quality and quantity of NK cells produced from these NK progenitor cells.

Keywords: CAR-NK cell; NK cell; adoptive cell therapy; cord blood; progenitor cell.