Interrogating the in vivo pharmacokinetics of armored CARs with radiohapten capture

通过放射性半抗原捕获来探究装甲 CAR 的体内药代动力学

基本信息

批准号：
10447176
负责人：
Simone Krebs
金额：
$ 55.17万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-07 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10447176
关键词：
Ablation Adoptive Cell Transfers Affinity Animals Antibodies Antigen Receptors Antigens Autoradiography B lymphoid malignancy Binding Biodistribution Biological Biological Assay CAR T cell therapy CD19 gene CD40 Antigens CD40 Ligand Cell Count Cell surface Cells Clinical Clinical Oncology Supplement (K12)Clinical Trials Complex Dose Drug Kinetics Ensure Excretory function Exhibits Generations Goals Haptens Hepatobiliary Human Image Immune ImmunoPET Immunohistochemistry In Vitro Inflammatory Inflammatory Response Pathway Kinetics Lanthanoid Series Elements Location Malignant Neoplasms Methodology Methods Modeling Molecular Target Monitor Mus Pathway interactions Patients Pharmaceutical Preparations Phenotype Play Positron-Emission Tomography Proteus Radiation Toxicity Radiometry Radionuclide therapy Relapse Reporter Reporter Genes Role Safety Series Specificity System T cell therapy T-Lymphocyte Therapeutic Therapy trial Time Tissues Titrations Translations Treatment Efficacy Treatment Failure Variant Work Xenograft Model Xenograft procedure base cell killing cellular imaging chimeric antigen receptor chimeric antigen receptor T cells clinical application clinical translation clinically relevant contrast imaging cytotoxic dosimetry effector T cell engineered T cells exhaustion gene function improved in vivo in vivo monitoring insight method development molecular imaging mouse model neoplastic cell next generation novel preclinical study preservation response single photon emission computed tomography success theranostics therapeutic target tool trafficking treatment effect tumor tumor heterogeneity tumor microenvironment uptake weapons

项目摘要

Project Summary/Abstract CD19-targeted chimeric antigen receptor (CAR) T cell therapy has shown remarkable treatment effects in B-cell malignancies, but many patients suffer from limited response and CD19-negative tumor relapse. Recently, we demonstrated that next-generation “armored” CAR T cells constitutively expressing the immune-stimulatory molecule CD40 ligand (CD40L) exhibited superior antitumor efficacy in preclinical studies but have not yet been fully explored in the context of CAR antigen-negative tumor relapse. There remains an urgent need for the non-invasive in vivo tracking of transfused T cells to determine their biodistribution, expansion, and functionality and to increase the CAR T cells’ killing capacity in case of imminent treatment failure. To overcome these limitations, we began developing methods for monitoring the in vivo kinetics of CAR T cells based on the concept of immune cell radiohapten capture. We demonstrated for the first time that T cells can be successfully transduced with a DOTA-antibody reporter, the DAbR1, enabling their in vivo tracking via PET and SPECT. In the current proposal, we build on this work and optimize our approach for translation of immune cell radiohapten capture from animals to patients, based on greatly improved components of 1) radiohapten capture reporter scFv C825 with picomolar binding affinity, and 2) optimized radiohaptens, the next- generation Proteus-DOTA (Pr) series suitable for imaging and targeted alpha therapy (TAT). The primary objectives of this study are to develop a clinically applicable PET imaging strategy of CAR T cell trafficking in B- cell malignancies and further study the effect of CD40L on counteracting the immune inhibition in syngeneic models of B-cell malignancies. The secondary objectives are to deliver TAT to enhance T cells’ killing capacity in cases of imminent treatment failure and improve the potency of CAR T cell therapies. We project to achieve our aims by generating second-generation and armored CD 19 CAR T cells expressing cell-surface anchored scFv C825. Syngeneic and immunodeficient xenograft murine models of CD19+ B cell malignancies including antigen-loss variants will be employed. After successful transduction, we will assess in vitro functionality of the CAR and the reporter, as well as radiation toxicity, followed by in vivo functionality, imaging sensitivity, and biodistribution, and develop an armored CAR T cell-based theranostic approach with the novel pair [86Y]YPr/[225Ac]AcPr. Finally, as a prerequisite to clinical translation of this novel platform, we will conduct studies using human second-generation and armored CAR T cells in clinically relevant xenograft models. The availability of such a single platform would provide crucial information for safer, more effective clinical trials. The aims thus have immediate translational relevance for our current clinical CD19 CAR T studies and other planned CAR T cell therapy trials.

项目概要/摘要 CD19靶向嵌合抗原受体（CAR）T细胞疗法在B细胞中显示出显着的治疗效果恶性肿瘤，但许多患者的反应有限且 CD19 阴性肿瘤复发。证明下一代“装甲”CAR T 细胞组成型表达免疫刺激分子CD40配体（CD40L）在临床前研究中显示出优异的抗肿瘤功效，但尚未得到证实在 CAR 抗原阴性肿瘤复发的背景下进行了充分探索。仍然迫切需要对输注的 T 细胞进行非侵入性体内追踪以确定其生物分布、扩增和功能，并在紧急情况下提高 CAR T 细胞的杀伤能力为了克服这些限制，我们开始开发监测体内动力学的方法。我们首次证明了基于免疫细胞放射性半抗原捕获概念的 CAR T 细胞。 T 细胞可以用 DOTA 抗体报告基因 DAbR1 成功转导，从而实现体内追踪通过 PET 和 SPECT，我们在这项工作的基础上优化了我们的翻译方法。从动物到患者的免疫细胞放射性半抗原捕获，基于大大改进的成分 1) 具有皮摩尔结合亲和力的放射性半抗原捕获报告基因 scFv C825，以及 2) 优化的放射性半抗原，下一个- 新一代 Proteus-DOTA (Pr) 系列适用于成像和靶向 α 治疗 (TAT)。本研究的目标是开发一种临床适用的 CAR T 细胞运输 B 细胞 PET 成像策略细胞恶性肿瘤并进一步研究CD40L对抗同基因免疫抑制的作用 B 细胞恶性肿瘤模型的次要目标是提供 TAT 以增强 T 细胞的杀伤能力。在治疗即将失败的情况下，提高 CAR T 细胞疗法的效力。我们计划实现我们的目标是生成表达细胞表面锚定的第二代装甲 CD 19 CAR T 细胞 CD19+ B 细胞恶性肿瘤的同基因和免疫缺陷异种移植小鼠模型，包括将采用抗原丢失变体。成功转导后，我们将评估转导的体外功能 CAR 和报告基因，以及辐射毒性，其次是体内功能、成像灵敏度和生物分布，并开发一种基于装甲 CAR T 细胞的治疗诊断方法 [86Y]YPr/[225Ac]AcPr 最后，作为这个新平台转化临床的先决条件，我们将进行研究。在临床相关的异种移植模型中使用人类第二代和装甲 CAR T 细胞。这样一个单一平台的建立将为更安全、更有效的临床试验提供重要信息。与我们当前的临床 CD19 CAR T 研究和其他计划的 CAR T 具有直接的转化相关性细胞治疗试验。