De novo designed receptor transmembrane domains (TMDs) present opportunities for precise control of cellular receptor functions. We developed a de novo design strategy for generating programmed membrane proteins (proMPs): single-pass α-helical TMDs that self-assemble through computationally defined and crystallographically validated interfaces. We used these proMPs to program specific oligomeric interactions into a chimeric antigen receptor (CAR) and found that both CAR T cell cytokine release and in vivo antitumor activity scaled linearly with the oligomeric state encoded by the receptor TMD. All programmed CARs (proCARs) stimulated substantially lower T cell cytokine release relative to the commonly used CD28 TMD, which we show elevated cytokine release through lateral recruitment of endogenous T cell costimulatory receptor CD28. Precise design using orthogonal and modular TMDs thus provides a new way to program receptor structure and predictably tune activity for basic or applied synthetic biology.

中文翻译：

---

从头设计的跨膜结构域调节工程受体功能

从头设计的受体跨膜结构域 (TMD) 为精确控制细胞受体功能提供了机会。我们开发了一种用于生成程序化膜蛋白 (proMP) 的从头设计策略：通过计算定义和晶体学验证界面自组装的单程 α 螺旋 TMD。我们使用这些 proMP 将特定的寡聚相互作用编程到嵌合抗原受体 (CAR) 中，并发现 CAR T 细胞细胞因子的释放和体内抗肿瘤活性与受体 TMD 编码的寡聚状态呈线性关系。相对于常用的 CD28 TMD，所有程序化的 CAR（proCAR）刺激了显着降低的 T 细胞细胞因子释放，我们通过内源性 T 细胞共刺激受体 CD28 的横向募集显示出细胞因子释放增加。