Engineering the ER Stress Response to Promote the Survival and Cytotoxic Specificity of CAR T cells

改造内质网应激反应以促进 CAR T 细胞的存活和细胞毒性特异性

• 批准号: 10212964
• 负责人: Samuel Robert Kerr
• 金额: $ 4.6万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212964
• 关键词: Acute Lymphocytic Leukemia; Adoptive Cell Transfers; Antibodies; Antigens; Apoptosis; Apoptotic; B-Cell Leukemia; B-Lymphocytes; Behavior Control; Binding; Biological; Biology; CD28 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; CTLA4 gene; Cell Survival; Cell membrane; Cells; Cleaved cell; Critical Pathways; Engineering; Equilibrium; Exposure to; Generations; Genetic Transcription; Human; In Vitro; Infection; Inflammatory; Introns; Isogenic transplantation; Leukocytes; Ligands; Logic; Malignant Neoplasms; Mediating; Messenger RNA; Mus; Pathway interactions; Patients; Plasma Cells; Post-Transcriptional Regulation; Proteins; RNA Splicing; Refractory; Relapse; Signal Pathway; Signal Transduction; Solid Neoplasm; Specificity; Speed; Stress; Surface; System; T cell differentiation; T-Cell Activation; T-Cell Antigen Receptor Specificity; T-Lymphocyte; TNFSF10 gene; Testing; Tissues; Transcript; Transcriptional Regulation; Work; XBP1 gene; biological adaptation to stress; cancer cell; cancer therapy; cell killing; chimeric antigen receptor; chimeric antigen receptor T cells; cytotoxic; cytotoxicity; effector T cell; endoplasmic reticulum stress; engineered T cells; eosinophil; genetic manipulation; improved; in vivo; macrophage; melanoma; mouse model; neoplastic cell; novel; response; tool; transcription factor; tumor; tumor microenvironment; tumor specificity; tumor-immune system interactions

PROJECT SUMMARY/ABSTRACT Chimeric antigen receptor (CAR) T cells have revolutionized cancer therapies and show incredible responses in patients with relapsed or refractory B cell cancers, yet are ineffective in solid tumors. In order to work in solid tumors, CAR T cells must be able to survive the immunosuppressive tumor microenvironment (TME) and distinguish between antigens presented on tumor cells and healthy tissue. Both of these obstacles can be overcome through clever synthetic engineering of the T cell using known biological circuits. We have identified a pathway critical to the survival and function of multiple types of leukocytes and naturally turned on during activation of CD8+ T cells, however surprisingly, it is not activated in the tumor microenvironment. Here, I propose to synthetically activate this pathway in order to promote CD8+ CAR T cell cytotoxicity and on-target, on-tumor specificity. Specifically, I will first explore multiple aspects of this signaling pathway and its ability to promote CAR T cell survival and cytotoxicity under the stresses in the tumor microenvironment. As a second strategy to improve CAR-T cell specificity for solid tumors, I will engineer a construct that selectively activates a CAR in response to post-transcriptional regulation. Benefits of post-transcriptional regulation include speed of activation, and amplification of signal. By making both the post-transcriptional regulator and the target dependent on binding target cell antigens, I will functionally create an “AND” logic gate, requiring stimulation from 2 separate antigens in order to facilitate CAR T cell killing. For both strategies I will first demonstrate their efficacy in vitro with primary mouse and human cells simulating TME conditions, and then in vivo using a syngeneic transplanted melanoma mouse model. Overall, I hypothesize that synthetic activation of this critical T cell pathway will enhance CAR-T behavior and control in the solid TME.

项目摘要/摘要 嵌合抗原受体（CAR）T细胞已彻底改变癌症疗法，并显示出令人难以置信的 继电器或难治性B细胞癌患者的反应，但在实体瘤中却无效。为了 在实体瘤中工作，CAR T细胞必须能够在免疫抑制肿瘤微环境中生存 （TME）并区分肿瘤细胞和健康组织上的抗原。这两个障碍 可以使用已知的生物电路对T细胞的巧妙合成工程来克服。我们有 确定了对多种类型白细胞生存和功能至关重要的途径，并且自然打开 在激活CD8+ T细胞的过程中，无论多么令人惊讶，它在肿瘤微环境中没有被激活。这里， 我建议通过综合激活这一途径，以促进CD8+ CAR T细胞细胞毒性和靶标的，即 肿瘤特异性。具体来说，我将首先探索此信号通路的多个方面及其能力 在肿瘤微环境中的应力下促进CAR T细胞的存活和细胞毒性。一秒钟 改善实体瘤CAR-T细胞特异性的策略，我将设计一种选择性激活A的构造 响应于转录后调节的汽车。转录后法规的好处包括 激活和信号的扩增。通过制作后期调节器和目标 取决于结合靶细胞抗原，我将创建一个“和”逻辑门，需要刺激 从2个单独的抗原中促进杀死CAR T细胞。对于这两种策略，我将首先证明他们 在体外与原代小鼠和人类细胞模拟TME条件，然后在体内使用A 合成性移植黑色素瘤小鼠模型。总的来说，我假设这种关键的合成激活 T细胞通路将增强固体TME中的CAR-T行为和控制。