Precision quality check of immunotherapeutics via single-cell cytokine mapping

通过单细胞细胞因子图谱对免疫治疗进行精确质量检查

• 批准号: 9518723
• 负责人: Timothy S McConnell
• 金额: $ 91.05万
• 依托单位: ISOPLEXIS, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-21 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9518723
• 关键词: Address; Adoptive Transfer; Antibodies; Antigens; Area; Autoimmune Process; Autologous; Automation; Biological Assay; Businesses; CD19 Antigens; CD19 gene; Cancer Patient; Cell Therapy; Cell physiology; Cells; Cellular immunotherapy; Clinical; Clinical Data; Clinical Trials; Collaborations; Cytometry; Data; Devices; Ensure; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Goals; Gold; Grant; HIV; Hematologic Neoplasms; Heterogeneity; Image; Immune; Immunosuppressive Agents; Immunotherapeutic agent; Immunotherapy; Individual; Informatics; Infusion procedures; Legal patent; Life; Lymphoma; Measures; Memorial Sloan-Kettering Cancer Center; Neurologic; Pancreas; Patients; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phenotype; Population; Proteins; Proteomics; Protocols documentation; Receptor Cell; Safety; Sample Size; Sampling; Site; Slide; Small Business Innovation Research Grant; Solid Neoplasm; Staging; Surface; System; T cell therapy; T-Lymphocyte; Technology; Testing; Time; Toxic effect; Treatment Efficacy; base; cellular engineering; chimeric antigen receptor; chromatin immunoprecipitation; cohort; cytokine; cytokine release syndrome; design; immunotherapy trials; immunotoxicity; improved; leukemia/lymphoma; member; patient response; phenotypic biomarker; predictive marker; response; tumor

Adoptive transfer of autologous T cells engineered to express chimeric antigen receptors (CARs) has emerged as a promising immunotherapy for patients with hematologic malignancies, such as CD19 CARs in leukemias and lymphomas. However, challenges remain in terms of manufacturing consistency and the functional profile of the CAR-T cell product, since infused cells, as “living drugs”, can be activated and release a variety of cytokines upon specific antigen recognition. These secreted cytokines may result in not only therapeutic efficacy, but also life-threatening immunotoxicity, such as cytokine release syndrome and neurologic toxicity. It is essential for cancer patients to have a full spectrum potency and toxicity profiling of CAR-T cell products before infusion. We have developed a single-cell barcode chip (SCBC) technology, which enables (a) assaying 30+ secreted proteins per individual live cell, (b) a broad range of immune cell functions covering efficacy and safety, (c) is designed to fit various types of immune cells, (d) is the first quantitative metric of these proteins per cell, and (e) requires samples sizes totaling only 5000 cells, all critical leaps over flow cytometry based platforms. With the collaboration of two leading CAR-T pharmaceutical companies, IsoPlexis’ single-cell deep profiling has revealed, for the first time, a pre-infusion correlate to post-infusion patient response. The SCBC analysis of 20 patients with lymphoma demonstrates a significant correlation of the polyfunctional strength of the CAR-T cells with objective response (complete or partial) to the CAR-T cell therapy (p = 0.012), indicating a powerful metric of quality assessment, where the other gold standard technologies did not detect any significant correlations. Based on the clinical data and the automation progress in our SBIR Phase I grant, we are proposing the following specific aims in this two-year Phase II application: AIM 1: Develop automated “flow cell” consumable compartment, which captures 32-plex single-cell cytokine response, to prepare for full automation required by large CAR-T trial (months 1-10); AIM 2: Produce a fully automated cartridge analysis and workflow system, to allow 10 samples assayed in parallel with minimal user interaction, allowing easy introduction into clinical core labs (months 1-15); AIM 3: Produce a comprehensive informatics suite, and fully test our automated system and informatics in two 30 patient cohort trials, providing robust and predictive biomarkers for pre-infusion CAR-T quality. Establish MSKCC and UCLA beta sites for post-phase II transition (months 7-24). With this Phase II submission, we will deliver the first effective CAR-T pre-infusion quality check assay to predict objective response in patients, in an easy-to-use, automated system that can be used throughout all cellular immunotherapy trials.

为表达嵌合抗原受体（CAR）设计的自体T细胞的收养转移已经出现 作为血液系统恶性肿瘤患者的有希望的免疫疗法，例如白血病中的CD19汽车 和淋巴瘤。但是，在制造一致性和功能概况方面仍然存在挑战 CAR-T细胞产物的中，由于感染的细胞作为“活着的药物”可以被激活并释放多种 特定抗原识别的细胞因子。这些分泌的细胞因子不仅会导致治疗效率，还会导致 而且还威胁生命的免疫毒性，例如细胞因子释放综合征和神经毒性。这是必不可少的 癌症患者在输注前具有全光谱效力和CAR-T细胞产物的毒性分析。 我们已经开发了一种单细胞条形码芯片（SCBC）技术，该技术使（a）分析30多个分泌 每个单个活细胞的蛋白质，（b）涵盖效率和安全性的广泛的免疫电池功能，（c）为 （d）旨在适合各种类型的免疫细胞，是每个细胞这些蛋白质的第一个定量度量，（e） 需要总计仅5000个单元的样品尺寸，所有关键飞跃都超过了基于流式细胞仪的平台。 通过两家领先的CAR-T制药公司的合作，Isoplexis的单细胞深处分析有 首次揭示了灌注前与输注后患者的反应相关。 SCBC分析20 患有淋巴瘤的患者表现出CAR-T细胞多功能强度的显着相关性 对CAR-T细胞疗法的客观反应（完全或部分）（p = 0.012），表明强大的度量 质量评估，其他黄金标准技术未检测到任何显着相关性。 根据我们SBIR I期授予的临床数据和自动化进度，我们提出以下内容 在这两年的II阶段应用中的具体目标：目标1：开发自动化的“流动单元” 捕获32个PLEX单细胞细胞因子反应的隔室为准备完整的自动化做准备 大型CAR-T试验（1-10个月）；目标2：生成全自动墨盒分析和工作流系统， 允许与最少用户互动并行分配的10个样本，从而轻松引入临床核心 实验室（1-15个月）；目标3：制作全面信息套件，并全面测试我们的自动化系统， 在两项30项患者队列试验中的信息，为输注前CAR-T提供了强大的预测生物标志物 质量。建立MSKCC和UCLAβ地点，以进行后期II后转变（第7-24个月）。 通过此II阶段提交，我们将提供第一个有效的CAR-T前输注质量检查测定法，以预测 患者的客观反应，在易于使用的自动化系统中，可以在所有细胞中使用 免疫疗法试验。