Adrenergic modulation of cellular immune functions in CAR T cell-induced cytokine release syndrome

CAR T 细胞诱导的细胞因子释放综合征中细胞免疫功能的肾上腺素调节

基本信息

批准号：
10532157
负责人：
Renyuan Bai
金额：
$ 61.39万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10532157
关键词：
Adrenergic Agents Adrenergic Receptor Antitumor Response B lymphoid malignancy Biological Assay CD14 gene CD19 gene CRISPR/Cas technology Cancer Patient Catecholamines Cell Therapy Cells Clinical Coculture Techniques Data Dedications Development Epinephrine Functional disorder Genetic Goals Granulocyte-Macrophage Colony-Stimulating Factor Hematologic Neoplasms Human IL6 gene Immune Immune response Immunosuppression Immunotherapy In Vitro Individual Inflammatory Institution Interleukin-6 Knock-out Knockout Mice Laboratories Life Macrophage Macrophage Activation Malignant Neoplasms Measures Mediating Mediator Modeling Molecular Mus Myelogenous Myeloid Cells NOS2A gene Pathogenesis Patient Monitoring Patient-Focused Outcomes Patients Pennsylvania Peripheral Blood Mononuclear Cell Prazosin Production Productivity Receptor Signaling Research Role Safety Sampling Scientist Serum Severities Shapes Solid Neoplasm Supportive care Symptoms T cell response T cell therapy T-Lymphocyte TNFSF5 gene Therapeutic Toxic effect Tyrosine 3-Monooxygenase United States National Institutes of Health alpha-Methyltyrosine anakinra antagonist cancer therapy chimeric antigen receptor chimeric antigen receptor T cells clinical implementation cytokine cytokine release syndrome data sharing effective intervention immune activation immune function immune-related adverse events immunotoxicity improved in vivo in vivo evaluation indexing inhibitor knock-down leukemia mortality mouse model neoplastic cell novel pharmacologic prevent receptor response small hairpin RNA success therapy outcome tool translational potential tumor tumor eradication

项目摘要

Immunotherapy using tumor-directed chimeric antigen receptor (CAR)-expressing T cells (CART) is a rapidly- emerging, promising cancer therapy that can elicit rapid, durable clinical responses, seen in anti-CD19 CART (CART19)-treated CD19+ B-cell malignancies. However, CART-evoked severe or fatal immune-related adverse events (irAE) including cytokine-release syndrome (CRS) are significant clinical barriers and safety concerns that impede CART use in cancer patients, even with strict patient monitoring and supportive care. Current tools for mitigating such immunotoxicities/CRS target individual cytokines/receptors (e.g. IL-6R, IL-1Ra, GM-CSF) or nonspecific immunosuppression, but preventing and treating such toxicity will require better understanding of cellular/molecular mediators of CART-induced CRS. This project aims to overcome these barriers and enable more effective interventions against immunotoxicities, and thus more effective CART treatments, building upon the PIs' novel, potentially translatable findings of a key role of catecholamines (CATs) in shaping CART therapy- associated immune activation. We found that blocking CAT synthesis both reduced CRS during CART therapy in mice and also enhanced its tumor eradication, and also found that CAT levels are elevated in patients with CART-evoked CRS, and correlated with symptoms and IL-6 levels, suggesting CATs contribute to human CRS pathophysiology. The project's goal is to identify the mechanistic CRS-modulating roles of these novel CAT actions and their impacts on macrophages and anti-tumor responses, and assess translational potential for improving CART cancer therapy outcomes using human patient samples. Specific Aims are: 1) Determine the mediating α1-AR subtype and its role in modulating CART-evoked cytokine release and anti-tumor responses in vitro and in vivo, based on our finding that CAT-associated cytokine release is mediated by α1-adrenergic receptors (α1-AR), of as-yet unknown subtype(s). We will identify the α1-AR subtype(s) that mainly mediate(s) CAT-evoked cytokine production in macrophages and CART, using in vitro co-culture assays and a mouse CART therapy model via genetic knockout (KO) and knockdown (KD) of α1-AR subtypes. 2) Determine the impact of myeloid-derived CATs on cytokine release and myeloid function during CART-therapy in vitro and in a mouse model, by inactivating CAT synthesis in myeloid cells via tyrosine hydroxylase (TH) KO, and assess its impact on cytokine release and macrophage function in a co-culture assay and in vivo using a LysM-Cre-mediated TH KO mouse. 3) Assess CAT induction in human patients during CART therapy, and CATs' dual impacts on irAE and tumor responses, and potential utility of CAT measures and blockers in predicting and preventing CRS, by quantifying circulating CAT and cytokine levels in patient serum samples collected during CART therapy, and evaluating correlations of CAT levels with clinical and laboratory CRS indices, anti-tumor responses and survival.

使用表达肿瘤的嵌合抗原受体 (CAR) 的 T 细胞 (CART) 进行免疫治疗是一种快速的治疗方法。新兴的、有前途的癌症疗法，可以引发快速、持久的临床反应，如抗 CD19 CART 所示 (CART19) 治疗的 CD19+ B 细胞恶性肿瘤然而，CART 会引起严重或致命的免疫相关不良反应。包括细胞因子释放综合征 (CRS) 在内的事件 (irAE) 是重大的临床障碍和安全问题即使有严格的患者监测和支持性护理，这也会阻碍 CART 在癌症患者中的使用。用于减轻此类免疫毒性/CRS 靶向个体细胞因子/受体（例如 IL-6R、IL-1Ra、GM-CSF）或非特异性免疫抑制，但预防和治疗此类毒性需要更好地了解 CART 诱导的 CRS 的细胞/分子介质该项目旨在克服这些障碍并实现这一目标。更有效的针对免疫毒性的干预措施，从而更有效的 CART 治疗 PIs关于儿茶酚胺（CAT）在塑造CART疗法中的关键作用的新颖的、潜在可转化的发现- 我们发现，在 CART 治疗期间，阻断 CAT 合成均可减少 CRS。在小鼠中也增强了其肿瘤根除能力，并且还发现 CAT 水平在患有以下疾病的患者中升高 CART 诱发 CRS，并与症状和 IL-6 水平相关，表明 CAT 有助于人类 CRS 该项目的目标是确定这些新型 CAT 的 CRS 调节作用。作用及其对巨噬细胞和抗肿瘤反应的影响，并评估转化潜力使用人类患者样本改善 CART 癌症治疗结果的具体目标是： 1) 确定介导α1-AR亚型及其在调节CART诱发的细胞因子释放和抗肿瘤反应中的作用体外和体内，基于我们的发现，CAT 相关细胞因子的释放是由 α1-肾上腺素能介导的受体 (α1-AR)，其亚型尚不清楚我们将鉴定主要介导的 α1-AR 亚型。使用体外共培养测定和小鼠 CART，巨噬细胞和 CART 中 CAT 诱发的细胞因子产生通过 α1-AR 亚型的基因敲除 (KO) 和敲低 (KD) 的治疗模型 2) 确定影响。骨髓源性 CAT 对体外和小鼠 CART 治疗期间细胞因子释放和骨髓功能的影响模型，通过酪氨酸羟化酶 (TH) KO 灭活骨髓细胞中的 CAT 合成，并评估其影响使用 LysM-Cre 介导的 TH 在共培养测定中和体内对细胞因子释放和巨噬细胞功能的影响 3) 评估 CART 治疗期间人类患者的 CAT 诱导，以及 CAT 对 irAE 的双重影响。和肿瘤反应，以及 CAT 措施和阻断剂在预测和预防 CRS 方面的潜在效用，量化 CART 治疗期间收集的患者血清样本中的循环 CAT 和细胞因子水平，以及评估 CAT 水平与临床和实验室 CRS 指数、抗肿瘤反应和生存的相关性。