Engineered human pluripotent stem cell-derived natural killer cells with PD-L1 responsive immunological memory for enhanced immunotherapeutic efficacy

• Published in: Bioactive materials Vol. 27; pp. 168 - 180
• Main Authors: Chang, Yun, Jin, Gyuhyung, Luo, Weichuan, Luo, Qian, Jung, Juhyung, Hummel, Sydney N., Torregrosa-Allen, Sandra, Elzey, Bennett D., Low, Philip S., Lian, Xiaojun Lance, Bao, Xiaoping
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.09.2023; KeAi Publishing; KeAi Communications Co., Ltd
• Subjects: Chimeric antigen receptor; Human pluripotent stem cells; Immunological memory; Immunotherapy; Natural killer cells; Immunological memory; Natural killer cells; Chimeric antigen receptor; Human pluripotent stem cells; Immunotherapy
• ISSN: 2452-199X; 2452-199X
• DOI: 10.1016/j.bioactmat.2023.03.018

Adoptive chimeric antigen receptor (CAR)-engineered natural killer (NK) cells have shown promise in treating various cancers. However, limited immunological memory and access to sufficient numbers of allogenic donor cells have hindered their broader preclinical and clinical applications. Here, we first assess eight different CAR constructs that use an anti-PD-L1 nanobody and/or universal anti-fluorescein (FITC) single-chain variable fragment (scFv) to enhance antigen-specific proliferation and anti-tumor cytotoxicity of NK-92 cells against heterogenous solid tumors. We next genetically engineer human pluripotent stem cells (hPSCs) with optimized CARs and differentiate them into functional dual CAR-NK cells. The tumor microenvironment responsive anti-PD-L1 CAR effectively promoted hPSC-NK cell proliferation and cytotoxicity through antigen-dependent activation of phosphorylated STAT3 (pSTAT3) and pSTAT5 signaling pathways via an intracellular truncated IL-2 receptor β-chain (ΔIL-2Rβ) and STAT3-binding tyrosine-X-X-glutamine (YXXQ) motif. Anti-tumor activities of PD-L1-induced memory-like hPSC-NK cells were further boosted by administering a FITC-folate bi-specific adapter that bridges between a programmable anti-FITC CAR and folate receptor alpha-expressing breast tumor cells. Collectively, our hPSC CAR-NK engineering platform is modular and could constitute a realistic strategy to manufacture off-the-shelf CAR-NK cells with immunological memory-like phenotype for targeted immunotherapy. [Display omitted] •Off-the-shelf dual CAR-NK cells are derived from hPSCs.•Optimized NK-specific CAR constructs enhance NK cell proliferation and function.•Universal FITC-CAR enhances antitumor activity of hPSC-derived NK cells.•Dual CAR hPSC-NK cells demonstrate improved antigen-responsive in vivo expansion.