Engineering synthetic helper cells that autonomously deliver orthogonal IL-2 to selectively promote therapeutic T cell proliferation in tumors

项目摘要/摘要 LIM/UCSF和六月/宾夕法尼亚大学IOTN中心之间的合作目标是开发一种新的 自主和选择性地驱动实体瘤内CAR T细胞增殖的方法。当前临床 数据表明，强大的增殖是过继转移T细胞治疗成功的关键1，2。 实体瘤这是一个特别严峻的挑战，因为肿瘤细胞被一种免疫抑制剂包围着。 微环境对T细胞的扩增和活性有多种障碍，包括抑制 调节性T细胞作为促增殖细胞因子3以及其他抑制细胞和因子的汇 促进终末T细胞分化进入耗尽状态4，5。为了解决这些问题，我们将结合 我们两个小组的专业知识：加西亚实验室(六月/宾夕法尼亚大学中心)在工程正交方面拥有专业知识 细胞因子/受体系统，而LIM实验室(LIM/UCSF中心)在工程合成助手方面拥有专业知识 能够自主识别和有针对性地重塑肿瘤微环境的细胞 合成旁分泌细胞因子回路。在这项合作中，我们建议设计合成辅助细胞， 在肿瘤中自主产生正交的IL-26，从而选择性地驱动局部CAR T细胞的扩增。这 协作以协同的方式合并两个中心的方法。如果成功，这种方法将产生 一种比促进肿瘤中CAR T细胞扩增更有效、选择性和自主性的方法 在最初的个人中心目标内的方法。我们的具体目标是： 目的1.设计合成辅助性T细胞，向肿瘤微环境运送正向白细胞介素2 目的1.1：设计小鼠Oro-IL-2辅助/杀伤细胞系统；体外和同种肿瘤模型试验 目的1.2：设计人Oro-IL-2辅助/杀伤细胞系统；体外和在NSG异种移植瘤模型中进行测试 目的2：调节IL-2受体的正交性信号，以改善和调节细胞对邻位IL-2的反应。 系统地改变Oro-IL-2受体胞内结构域中的信号域以识别和部署 促进T细胞增殖和/或最大限度减少消耗的变种。

Project Summary/Abstract The goal of this collaboration between the Lim/UCSF and June/UPenn IOTN centers is to develop a novel approach to autonomously and selectively drive proliferation of CAR T cells within a solid tumor. Current clinical data indicates that strong proliferation is critical for therapeutic success of adoptively transferred T cells1,2. In solid tumors this is a particularly acute challenge as tumor cells are surrounded by an immunosuppressive microenvironment that presents multiple barriers to T cell expansion and activity, including suppressive regulatory T cells that act as sinks for pro-proliferation cytokines3, as well as other suppressor cells and factors that promote terminal T cell differentiation into an exhausted state4,5. To address these problems, we will combine the expertise of our two groups: the Garcia Lab (June/UPenn center) has expertise in engineering orthogonal cytokine/receptor systems, while the Lim Lab (Lim/UCSF center) has expertise in engineering synthetic helper cells that can autonomously recognize and remodel the tumor microenvironment in a targeted way using synthetic paracrine cytokine circuits. In this collaboration, we propose to engineer synthetic helper cells that autonomously produce orthogonal IL-26 in tumors, thereby selectively driving CAR T cell expansion locally. This collaboration merges approaches from both centers in a synergistic way. If successful, this approach will yield a way to promote CAR T cell expansion in tumors more effectively, selectively and autonomously than approaches within the original individual center aims. Our specific aims are to: Aim 1. Engineer synthetic helper T cells that deliver ortho-IL2 to the tumor micro-environment Aim 1.1: Engineer mouse ortho-IL-2 helper/killer cell system; test in vitro and in syngeneic tumor models Aim 1.2: Engineer human ortho-IL-2 helper/killer cell system; test in vitro and in NSG xenograft tumor models Aim 2: Modulate orthogonal IL-2 receptor signaling to improve and tune cell responses to ortho-IL2. Systematically alter the signaling domains in the ortho-IL-2 receptor intracellular domain to identify and deploy variants that increase T cell proliferation and/or minimize exhaustion.