Wibowo Tansri, Kogue Yosuke, Ikeda Shunya, Yaga Moto, Tachikawa Mana, Suga Makiko, Kida Shuhei, Shibata Kumi, Tsutsumi Kazuhito, Murakami Hiraku, Ueda Yasutaka, Kato Hisashi, Fukushima Kentaro, Fujita Jiro, Ueda Tomoaki, Kusakabe Shinsuke, Hino Akihisa, Ichii Michiko, Imai Chihaya, Okuzaki Daisuke, Kumanogoh Atsushi, Hosen Naoki
Department of Respiratory Medicine and Clinical Immunology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
Osaka Research Center for Drug Discovery, Otsuka Pharmaceutical Company, Ltd., Osaka 562-0029, Japan.
Mol Ther Methods Clin Dev. 2024 Nov 12;32(4):101374. doi: 10.1016/j.omtm.2024.101374. eCollection 2024 Dec 12.
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 , 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 culture for 2 weeks. Single-cell RNA sequencing analysis of CD56CD3CD14CD19 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 CD34HLA-DR cells. In addition, among the CD34HLA-DR cells, only CD7 cells could differentiate into NK cells. These results indicate that CD56CD7CD34HLA-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.
靶向CD19的脐血（CB）来源的嵌合抗原受体（CAR）-自然杀伤（NK）细胞已被证明对B细胞恶性肿瘤有效。虽然人CD56 NK细胞可以扩增，但NK细胞也可以从造血祖细胞分化而来。目前尚不清楚CAR-NK细胞是源自CB中的成熟NK细胞还是NK祖细胞。在这里，我们确定CAR-NK细胞主要源自CD56 NK祖细胞。我们首先发现，在培养2周后，大量的CD19 CAR-NK细胞从CD56 CB单核细胞中产生。对CD56CD3CD14CD19 CB单核细胞进行单细胞RNA测序分析表明，这些细胞可根据CD34和人类白细胞抗原（HLA）-DR的表达分为三个亚群。NK细胞主要源自CD34HLA-DR细胞。此外，在CD34HLA-DR细胞中，只有CD7细胞可以分化为NK细胞。这些结果表明，CD56CD7CD34HLA-DR谱系标志物（Lin）细胞是人类CB来源的CAR-NK细胞的主要来源，这表明开发提高从这些NK祖细胞产生的NK细胞质量和数量的方法具有重要意义。
