Inniss Mara C, Smith Sean G, Li Dan Jun, Primack Benjamin, Sun Dexue, Olinger Grace Y, Sheahan Kerri-Lynn, Ross Theresa, Langley Meghan, Young Violet, Alvarado Andres, Davoodi Shabnam, Geng Jiefei, Schebesta Michael, Ols Michelle L, Tchaicha Jeremy, Ter Meulen Jan, Sethi Dhruv K
Obsidian Therapeutics, Cambridge, MA, USA.
Commun Biol. 2025 Jan 9;8(1):28. doi: 10.1038/s42003-024-07410-z.
Adoptive cell therapies (ACT) have shown reduced efficacy against solid tumor malignancies compared to hematologic malignancies, partly due to the immunosuppressive nature of the tumor microenvironment (TME). ACT efficacy may be enhanced with pleiotropic cytokines that remodel the TME; however, their expression needs to be tightly controlled to avoid systemic toxicities. Here we show T cells can be armored with membrane-bound cytokines with surface expression regulated using drug-responsive domains (DRDs) developed from the 260-amino acid protein human carbonic anhydrase 2 (CA2). The CA2-DRD can be stabilized in vitro and in vivo with the FDA-approved small-molecule CA2 inhibitor acetazolamide (ACZ). We develop conditional degrons using library-based screening of mutants and show characterization of one DRD using crystallography and molecular dynamics (MD) simulations. Using protein-engineering solutions to increase the valency of DRDs fused to the cargo we have developed "modulation hubs" and show tight regulation of membrane-bound cytokines IL2, IL12, IL15, IL21, IL23, and IFNα in genetically engineered T cells. Finally, CA2-DRD regulated IL12 mediates regulated efficacy in a solid tumor model. Regulation of pleotropic cytokines potentially paves the way to safely use these powerful cytokines in ACT for cancer treatment.
与血液系统恶性肿瘤相比，过继性细胞疗法（ACT）对实体瘤恶性肿瘤的疗效有所降低，部分原因是肿瘤微环境（TME）的免疫抑制特性。多效性细胞因子可重塑TME，从而增强ACT的疗效；然而，它们的表达需要严格控制，以避免全身毒性。在这里，我们展示了T细胞可以配备膜结合细胞因子，其表面表达可通过使用从260个氨基酸的人类碳酸酐酶2（CA2）蛋白开发的药物反应结构域（DRD）来调节。CA2-DRD可以在体外和体内用FDA批准的小分子CA2抑制剂乙酰唑胺（ACZ）稳定下来。我们通过基于文库的突变体筛选开发了条件性降解子，并使用晶体学和分子动力学（MD）模拟展示了一个DRD的特征。通过蛋白质工程解决方案增加与货物融合的DRD的价态，我们开发了“调节枢纽”，并展示了在基因工程T细胞中对膜结合细胞因子IL2、IL12、IL15、IL21、IL23和IFNα的严格调控。最后，CA2-DRD调节的IL12在实体瘤模型中介导了调节后的疗效。多效性细胞因子的调控可能为在ACT中安全使用这些强大的细胞因子进行癌症治疗铺平道路。
