Department of Pharmacology, Penn State College of Medicine, Hershey, PA 17033.
Department of Biomedical Engineering, Pennsylvania State University, University Park, PA 16802.
Proc Natl Acad Sci U S A. 2024 Oct 29;121(44):e2405717121. doi: 10.1073/pnas.2405717121. Epub 2024 Oct 23.
Chimeric antigen receptor T cell therapies have achieved great success in eradicating some liquid tumors, whereas the preclinical results in treating solid tumors have proven less decisive. One of the principal challenges in solid tumor treatment is the physical barrier composed of a dense extracellular matrix, which prevents immune cells from penetrating the tissue to attack intratumoral cancer cells. Here, we improve immune cell infiltration into solid tumors by manipulating septin-7 functions in cells. Using protein allosteric design, we reprogram the three-dimensional structure of septin-7 and insert a blue light-responsive light-oxygen-voltage-sensing domain 2 (LOV2), creating a light-controllable septin-7-LOV2 hybrid protein. Blue light inhibits septin-7 function in live cells, inducing extended cell protrusions and cell polarization, enhancing cell transmigration efficiency through confining spaces. We genetically edited human natural killer cell line (NK92) and mouse primary CD8 T-cells expressing the engineered protein, and we demonstrated improved penetration and cytotoxicity against various tumor spheroid models. Our proposed strategy to enhance immune cell infiltration is compatible with other methodologies and therefore, could be used in combination to further improve cell-based immunotherapies against solid tumors.
嵌合抗原受体 T 细胞疗法在消除某些液体肿瘤方面取得了巨大成功，而在治疗实体瘤方面的临床前结果则不那么确定。实体瘤治疗的主要挑战之一是由致密细胞外基质组成的物理屏障，它阻止免疫细胞穿透组织攻击肿瘤内的癌细胞。在这里，我们通过操纵细胞中 septin-7 的功能来改善免疫细胞对实体瘤的浸润。我们使用蛋白质变构设计，重新编程 septin-7 的三维结构，并插入一个蓝光响应的光氧电压感应结构域 2（LOV2），创建一个光可控的 septin-7-LOV2 杂合蛋白。蓝光抑制活细胞中 septin-7 的功能，诱导细胞突起和细胞极化的延长，增强细胞穿过限制空间的迁移效率。我们对人自然杀伤细胞系（NK92）和表达工程蛋白的小鼠原代 CD8 T 细胞进行了基因编辑，并证明了其对各种肿瘤球体模型的穿透和细胞毒性得到了改善。我们提出的增强免疫细胞浸润的策略与其他方法兼容，因此可以结合使用以进一步提高针对实体瘤的基于细胞的免疫疗法。
