The role of catecholamines in immunotoxicity and tumor response of adoptive T cell therapy in cancer

胶质母细胞瘤(GBM)是最常见的原发脑肿瘤，也是人类最致命的癌症之一。不是 治愈是存在的，而且只取得了有限的治疗进展。一个非常有前途的领域 免疫疗法是过继转移基因工程的患者来源的血液淋巴细胞 利用嵌合抗原受体基因(CARS)靶向并摧毁癌细胞，目前正在探索 治疗肾小球基底膜。使用CAR修饰的T细胞(CART)的关键挑战涉及剂量限制的免疫毒性 包括细胞因子释放综合征(CRS)和神经毒性。大多数研究的目的是改进 CART的疗效，同时毒性/不良反应的机制，创新的战略，其 管理，以及它们对抗肿瘤疗效的影响仍未得到充分研究。我最近发现 内源性儿茶酚胺在免疫中通过自我放大的前馈环路驱动CART诱导的CRS 细胞及其产物的抑制对CART治疗的Raji淋巴瘤荷瘤小鼠的保护作用 致死性CRS和增强的肿瘤根除，提示免疫毒性和抗肿瘤的不同途径 回应。这项提议的目的是阐明儿茶酚胺的调节机制。 CART治疗脑肿瘤和其他癌症的免疫毒性及其对抗肿瘤的影响 免疫活性癌症模型的反应。我的中心假设是儿茶酚胺促进了CART- 其药理抑制可提高CART治疗的安全性和肿瘤特异性杀伤力。我 我将在3个具体目标上测试这一想法：1)评估儿茶酚胺和CRS在 免疫活性CD19 B细胞ALL CART19模型的建立，并确定其抑制作用的双重影响 儿茶酚胺的产生对细胞因子释放和抗肿瘤反应的影响 CD19 B细胞ALL CART19模型中细胞因子的释放，该模型概括了人类CART19中的CRS 治疗；2)确定儿茶酚胺上调儿茶酚胺产生的信号通路 和细胞因子释放在小鼠CART19(MCART19)治疗中，以及这如何影响CART功能，通过使用 基因表达微阵列和途径报告芯片发现肾上腺素的贡献机制- 诱导儿茶酚胺和细胞因子的合成，并通过测定细胞因子的影响最显著 改变mCART19细胞的激活、扩增和细胞毒性；3)决定是否阻断内源性 儿茶酚胺合成降低CART诱导的全身和中枢神经系统免疫毒性并增强抗肿瘤作用 免疫活性小鼠GBM模型的反应。这一结果将促进对 促进免疫毒性的内源性途径，并可能改善CART治疗的结果 通过对其机制的新见解和新的治疗方法，管理和管理不良免疫毒性 儿茶酚胺阻断策略，其抑制多种细胞因子的范围比抗IL6R抗体更广泛 (Tocilizumab)。

Glioblastoma (GBM) is the most common primary brain tumor, and among the most lethal human cancers. No cure exists, and only limited treatment advances have been achieved. A highly promising area of immunotherapy is adoptive transfer of genetically engineered, patient-derived blood lymphocytes transfected with chimeric antigen receptor genes (CARs) to target and destroy cancer cells, currently being explored for treating GBM. Critical challenges to using CAR-modified T (CART) cells involve dose-limiting immunotoxicities including cytokine-release-syndrome (CRS) and neurological toxicities. Most research has aimed at improving CART efficacy, while the mechanisms of toxicities/adverse reactions, innovative strategies for their management, and their implications for anti-tumor efficacy remain under-explored. I recently discovered that endogenous catecholamines drive CART-induced CRS, via a self-amplifying feed-forward loop in immune cells, and that inhibiting their production protected CART-treated Raji lymphoma-bearing xenograft mice from lethal CRS and enhanced tumor eradication, suggesting separate pathways of immunotoxicity and anti-tumor response. The goal of this proposal is to elucidate the mechanism by which catecholamines mediate immunotoxicity in CART therapy in brain tumors and other cancers and assess their impact on the anti-tumor response in immunocompetent cancer models. My central hypothesis is that catecholamines promote CART- induced CRS and that its pharmacologic inhibition improves CART therapy safety and tumor-specific killing. I will test this idea in 3 Specific Aims: 1) Evaluate the induction of catecholamines and CRS in an immunocompetent CD19+ B cell ALL CART19 model, and determine the dual impact of suppressing catecholamine production on cytokine release and anti-tumor responses, by analyzing catecholamine and cytokine release in a CD19+ B cell ALL CART19 model, which recapitulates the CRS seen in human CART19 therapy; 2) Determine the signaling pathway by which catecholamines upregulate catecholamine production and cytokine release in mouse CART19 (mCART19) therapy and how this affects CART functionality, by using gene expression microarray and pathway reporter arrays to discover contributing mechanisms of epinephrine- induced catecholamine and cytokine synthesis and by determining the effects of cytokines most significantly altered on mCART19 cell activation, expansion and cytotoxicity; 3) Determine whether blocking endogenous catecholamine synthesis reduces CART-induced systemic and CNS immunotoxicity and improves anti-tumor responses in an immunocompetent mouse GBM model. The results will advance basic understanding of endogenous pathways contributing to immunotoxicity, and may enable improved outcomes of CART therapy and management of adverse immunotoxicities, via new insights into their mechanisms and a novel treatment strategy of catecholamine blockade, which inhibits multiple cytokines more broadly than anti-IL6R antibody (tocilizumab).