Redesigning the T cell: Using Synthetic Biology to Engineer Therapeutic Cells

描述（由申请人提供）：工程细胞有潜力成为强大的“智能”治疗剂，但我们缺乏系统的工程细胞框架来优化其功效并最小化其不良反应。例如，表达人工肿瘤导向受体（嵌合抗原受体- CARs）的T细胞在治疗某些白血病方面取得了意想不到的成功。但是这种工程细胞缺乏天然T细胞的精确性和控制力：脱靶细胞杀伤以及免疫系统的过度激活会导致大多数患者出现严重的不良反应。实现对这些工程T细胞的控制仍然是一个主要的挑战，使其成为一种安全可靠的治疗方法，可以扩展到更广泛的疾病。我们建议应用合成生物学的方法来系统地设计治疗性免疫细胞，使其具有更加定制和精确控制的行为。我们将运用我们的知识细胞信号工程模块化开关和电路，可用于控制这些治疗细胞。我们的项目将跨越两个互补的方法，由以下目标代表：目标1。开发用于CAR - T细胞精确控制的电路模块工具箱。我们将构建可与car结合的模块，以提高其特异性和安全性，包括：a.通过小分子或光远程控制T细胞。为医生提供控制移植治疗细胞活性的安全开关。b.可调反馈电路。通过调整剂量-反应阈值、动力学校正和反应幅度/持续时间来优化CAR - T细胞反应。c.多抗原触发活化。增加对癌细胞靶向的辨别。d.用户引导的细胞迁移。将T细胞定位到特定的作用位点，提高精度。e。

DESCRIPTION (provided by applicant): Engineered cells have the potential to serve as powerful "smart" therapeutic agents, but we lack a systematic framework for engineering cells to optimize their efficacy and minimize their adverse effects. For example, T cells expressing artificial tumor-directed receptors (Chimeric Antigen Receptors - CARs) have achieved unexpected success in treating certain leukemias. But the engineered cells lack the precision and control of native T cells: off-target cell killing as well as hyperactivation of the immune system lead to severe adverse effects in most patients. Achieving control over these engineered T cells remains a major challenge to making this a safe and reliable therapy that can be extended to a wider range of diseases. We propose to apply the approaches of synthetic biology to systematically engineer therapeutic immune cells with far more tailored and precisely controlled behaviors. We will apply our knowledge of cell signaling to engineer modular switches and circuits that can be used to control these therapeutic cells. Our project will span two complementary approaches, represented by the following aims: Aim 1. Develop toolbox of circuit modules for precision control of CAR T cells. We will construct modules that can be combined with CARs to increase their specificity and safety, including: a. Remote control of T cells via small molecules or light. Provide physicians with safety switches for controlling activiy of transplanted therapeutic cells. b. Tunable feedback circuits. Optimize CAR T cell responses by tuning dose-response thresholds, kinetic proofreading, and the amplitude/duration of response. c. Multi-antigen triggered activation. Increase discrimination of cancer cell targeting. d. User-guided cell migration. Target T cells to specific sites of action, increasing precision. e. Synthetic cell-cell communication. Engineer autocrine and multi-cell decision-making circuits. Aim 2. Refactoring T cells: identify submodules in the T cell response network that can be used to construct custom response programs. Current CARs replicate the native program of T cell activation, but in response to a novel input. We postulate that there are many alternative ways to plug into the T cell signaling network and to generate custom tailored responses. We will scan combinatorial libraries of rewired T cell signaling proteins to identify alternative control points that can be activated through induced complex assembly (light or drugs). This network-rewiring library should reveal modular subnetworks that can be used as building blocks to construct custom response programs, including: a. Separate inducible control of: i) T cell proliferation, ii) tumor cell killing, and iii) memory cell establishment b. Induction of alternatie cytokine response programs ideal for cancer treatment (e.g. induce Th1 program) c. Induction of custom "a la carte" hybrid cytokine response programs that are optimized for cancer treatment.