Towards Safer and More Effective CART Cell Therapy Through the Modulation of Myeloid Cytokines

项目摘要 尽管嵌合抗原受体T（CART）细胞疗法在治疗B细胞白血病中具有令人印象深刻的活性， 对于恶性肿瘤，治疗受到毒性发展的限制，包括细胞因子释放综合征（CRS） 和神经毒性，以及较低的持久反应率。CART细胞的体内扩增是一种有效的方法。 是其抗肿瘤活性以及毒性发展的关键决定因素。在过去的五年里， 我们的工作重点是解决与CART复杂性、毒性和耐药性相关的限制。我们 利用伴侣犬作为概念验证模型，以测试工程化同种异体人CART细胞作为 治疗自发性疾病，包括弥漫性大B细胞淋巴瘤（DLBCL）。我们改造了犬科动物 靶向CD 20的异种人CART细胞（T细胞受体α（TRAC）-和β-2微球蛋白（B2 M）- 破坏），开发和优化犬特异性淋巴细胞清除方案，并测试 异种CART细胞在健康的比格犬。然后，为了在体内真实的时间内追踪CART细胞的命运和功能，我们 开发并最近发表了钠碘同向转运体（NIS）作为检测CART细胞的平台的实用性 在小鼠模型中的扩增、运输和毒性。在这个应用程序中，我们建议利用我们的NIS报告 系统，新型犬淋巴细胞清除方案，并建立犬临床试验专业知识，以测试NIS+人类 伴侣犬的CART细胞。我们假设NIS+犬的18F-四氟硼酸盐（TFB）PET成像 靶向CD 20的异种人CART细胞是检测CART细胞排斥、扩增 和向肿瘤部位的运输。为了验证这一假设，我们设计了两个 具体目标。在目标1中，我们将研究NIS+ CART细胞的TFB-PET成像测量CART的效用 在患有DLBCL的犬患者中的扩增、排斥和贩运。我们将确定定量PET 信号（qPET）与外周血中CAR转基因的定量PCR（qPCR）测量相关 和淋巴结样本。在目标2中，我们将确定CART细胞扩增的TFB-PET成像以及 贩运与其活性和毒性有关。这些目标的完成将验证qPET成像作为一种 研究犬淋巴瘤中CART动力学的非侵入性平台，并提供额外的理由， NIS+ CART细胞在淋巴瘤患者中的首次人体临床试验。

PROJECT SUMMARY Despite the impressive activity of chimeric antigen receptor T (CART) cell therapy in the treatment of B-cell malignancies, the therapy is limited by the development of toxicities, including cytokine release syndrome (CRS) and neurotoxicity, as well as by lower rates of durable responses. The in vivo expansion of CART cells is a critical determinant of their antitumor activity as well as of the development of toxicities. Over the last 5 years, we have focused our work on addressing limitations related to CART complexity, toxicity, and resistance. We utilized companion dogs as a proof-of-concept model to test engineered allogeneic human CART cells as a therapy for spontaneous disease including diffuse large B-cell lymphoma (DLBCL). We engineered canine CD20-targeting xenogeneic human CART cells (T-cell receptor alpha (TRAC)- and beta-2 microglobulin (B2M)- disrupted), developed and optimized a canine-specific lymphodepleting regimen, and tested the safety of xenogeneic CART cells in healthy beagles. Then, to track CART cell fates and functions in real time in vivo, we developed and recently published the utility of sodium iodide symporter (NIS) as a platform to detect CART cell expansion, trafficking, and toxicity in mouse models. In this application, we propose to utilize our NIS reporter system, novel canine lymphodepletion regimen, and established canine clinical trial expertise to test NIS+ human CART cells in companion dogs. We hypothesize that 18F-tetrafluoroborate (TFB) PET imaging of NIS+ canine CD20-targeting xenogeneic human CART cells is a sensitive strategy to detect CART cell rejection, expansion, and trafficking to tumor sites in canine patients with DLBCL. To test this hypothesis, we have designed two specific aims. In Aim 1, we will study the utility of TFB-PET imaging of NIS+ CART cells to measure CART expansion, rejection, and trafficking in canine patients with DLBCL. We will determine how quantitative PET signal (qPET) correlates with quantitative PCR (qPCR) measurement of the CAR transgene in peripheral blood and lymph node samples. In Aim 2, we will determine how TFB-PET imaging of CART cell expansion and trafficking correlates with their activity and toxicity. Completion of these aims will validate qPET imaging as a non-invasive platform to study CART dynamics in canine lymphoma and provide additional rationale to propose a first-in-human clinical trial of NIS+ CART cells in patients with lymphoma.