Engineered human pluripotent stem cell-derived natural killer cells with PD-L1 responsive immunological memory for enhanced immunotherapeutic efficacy
Engineered human pluripotent stem cell-derived natural killer cells with PD-L1 responsive immunological memory for enhanced immunotherapeutic efficacy
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DOI：
10.1016/j.bioactmat.2023.03.018
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发表时间：
2023-09-01
影响因子：
18.9
通讯作者：
Bao, Xiaoping
中科院分区：
文献类型：
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作者：
Chang, Yun;Jin, Gyuhyung;Bao, Xiaoping
过继嵌合抗原受体（CAR）工程的自然杀伤细胞（NK）在治疗多种癌症方面显示出前景。然而，有限的免疫记忆和获得足够数量的同种异体供体细胞阻碍了它们更广泛的临床前和临床应用。在这里，我们首先评估了八种不同的CAR结构，它们使用抗pd - l1纳米体和/或通用抗荧光素（FITC）单链可变片段（scFv）来增强NK-92细胞对异质实体瘤的抗原特异性增殖和抗肿瘤细胞毒性。接下来，我们用优化的car对人类多能干细胞（hPSCs）进行基因工程改造，并将其分化为功能性的双CAR-NK细胞。肿瘤微环境响应性抗pd - l1 CAR通过细胞内截断IL-2受体β链（Delta IL-2R β）和STAT3结合酪氨酸- x -谷氨酰胺（YXXQ）基序，通过抗原依赖性激活磷酸化STAT3 （pSTAT3）和pSTAT5信号通路，有效促进hPSC-NK细胞增殖和细胞毒性。通过在可编程抗fitc CAR和表达叶酸受体α的乳腺肿瘤细胞之间架起桥梁的fitc -叶酸双特异性适配器，pd - l1诱导的记忆样hPSC-NK细胞的抗肿瘤活性进一步增强。总的来说，我们的hPSC CAR-NK工程平台是模块化的，可以构成一个现实的策略来制造现成的CAR-NK细胞，具有免疫记忆样表型，用于靶向免疫治疗。
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 acti-vation of phosphorylated STAT3 (pSTAT3) and pSTAT5 signaling pathways via an intracellular truncated IL-2 receptor beta-chain (Delta IL-2R beta) 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.