肿瘤细胞治疗研究
英文原题：Metabolic reprogramming via an engineered PGC-1α improves human chimeric antigen receptor T-cell therapy against solid tumors.
我们的数据进一步支持代谢重编程在免疫调节治疗中的作用，并凸显 PGC-1 等基因作为有吸引力的候选者，可与嵌合受体或 TCR 一同纳入载荷，用于实体瘤的细胞治疗。
背景：癌症细胞免疫疗法可通过向患者免疫系统补充大量肿瘤特异性T细胞增强免疫功能。嵌合抗原受体（CAR）疗法通过基因工程“重定向”外周T细胞，使其靶向肿瘤，在血液系统癌症中效力显著。然而，由于多种耐药机制，CAR-T疗法对实体瘤仍无效。我们及其他研究者发现，肿瘤微环境具有独特的代谢特征，形成免疫细胞功能障碍屏障。此外，肿瘤内T细胞分化异常会损害线粒体生物发生，导致严重的细胞内在代谢缺陷。虽然我们及其他研究者已证明，通过增强线粒体生物发生可改善小鼠T细胞受体（TCR）转基因细胞，本研究旨在确定是否可通过代谢重编程改善人CAR-T细胞功能。材料与方法：将抗EGFR CAR-T细胞输注至携带A549肿瘤的NSG小鼠体内，并分析TIL（肿瘤浸润淋巴细胞）的耗竭和代谢缺陷。采用携带PPARγ共激活因子1（PGC-1α）、PGC-1α S571A及NT-PGC-1α构建体的慢病毒，与抗EGFR CAR慢病毒共同转导T细胞。通过流式细胞术和Seahorse分析开展体外代谢分析，并进行RNA测序。最后，使用表达PGC-1α或NT-PGC-1α的抗EGFR CAR-T细胞治疗携带A549肿瘤的NSG小鼠，并分析PGC-1α共表达对肿瘤浸润CAR-T细胞的影响。结果：本研究显示，经过工程化改造、对抑制具有抗性的PGC-1α可重编程人CAR-T细胞代谢。PGC-1α转导CAR-T细胞的转录组分析表明，该方法有效诱导线粒体生物发生，也上调了与效应功能相关的程序。使用这些细胞治疗携带人实体瘤的免疫缺陷小鼠，显著提高了体内疗效。相比之下，截短型PGC-1α（NT-PGC-1α）未改善体内结局。结论：我们的数据进一步支持代谢重编程在免疫调节治疗中的作用，并凸显PGC-1α等基因作为载荷与嵌合受体或TCR共同用于实体瘤细胞治疗的潜力。
BACKGROUND: Cellular immunotherapies for cancer represent a means by which a patient's immune system can be augmented with high numbers of tumor-specific T cells. Chimeric antigen receptor (CAR) therapy involves genetic engineering to 'redirect' peripheral T cells to tumor targets, showing remarkable potency in blood cancers. However, due to several resistance mechanisms, CAR-T cell therapies remain ineffective in solid tumors. We and others have shown the tumor microenvironment harbors a distinct metabolic landscape that produces a barrier to immune cell function. Further, altered differentiation of T cells within tumors induces defects in mitochondrial biogenesis, resulting in severe cell-intrinsic metabolic deficiencies. While we and others have shown murine T cell receptor (TCR)-transgenic cells can be improved through enhanced mitochondrial biogenesis, we sought to determine whether human CAR-T cells could be enabled through a metabolic reprogramming approach. MATERIALS AND METHODS: Anti-EGFR CAR-T cells were infused in NSG mice which bore A549 tumors. The tumor infiltrating lymphocytes were analyzed for exhaustion and metabolic deficiencies. Lentiviruses carrying PPAR-gamma coactivator 1 (PGC-1 ), PGC-1 S571A and NT-PGC-1 constructs were used to co-transduce T cells with anti-EGFR CAR lentiviruses. We performed metabolic analysis via flow cytometry and Seahorse analysis in vitro as well as RNA sequencing. Finally, we treated therapeutically A549-carrying NSG mice with either PGC-1 or NT-PGC-1 anti-EGFR CAR-T cells. We also analyzed the differences in the tumor-infiltrating CAR-T cells when PGC-1 is co-expressed. RESULTS: Here, in this study, we show that an inhibition resistant, engineered version of PGC-1 , can metabolically reprogram human CAR-T cells. Transcriptomic profiling of PGC-1 -transduced CAR-T cells showed this approach effectively induced mitochondrial biogenesis, but also upregulated programs associated with effector functions. Treatment of immunodeficient animals bearing human solid tumors with these cells resulted in substantially improved in vivo efficacy. In contrast, a truncated version of PGC-1 , NT-PGC-1 , did not improve the in vivo outcomes. CONCLUSIONS: Our data further support a role for metabolic reprogramming in immunomodulatory treatments and highlight the utility of genes like PGC-1 as attractive candidates to include in cargo along with chimeric receptors or TCRs for cell therapy of solid tumors.
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