Precision Quality Check of Immunotherapeutics via Single Cell Cytokine Mapping

通过单细胞细胞因子图谱进行免疫治疗的精确质量检查

基本信息

批准号：
9202164
负责人：
Jing Zhou
金额：
$ 16.03万
依托单位：
ISOPLEXIS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-21 至 2017-03-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9202164
关键词：
Address Adoptive Transfer Adverse effects Antibodies Antigens Area Autoimmune Process Award B lymphoid malignancy B-Cell Lymphomas Binding Biological Assay CD19 Antigens CD19 gene Cell Therapy Cell physiology Cell secretion Cells Clinical Cytokine Activation Data Development Devices Ensure Flow Cytometry Future Gold Immune Immune response Immunotherapeutic agent In Vitro Inflammatory Infusion procedures Injection of therapeutic agent Interleukin-6 Learning Life Malignant Neoplasms Maps Marketing Measurement Measures Mediating Modification Monitor Nature Patients Pharmaceutical Preparations Pharmacologic Substance Phase Physicians Printing Proteins Protocols documentation Quality Control Reaction Reporting Reproducibility Risk Safety Sampling Slide Small Business Innovation Research Grant Surface Antigens Syndrome T cell therapy T-Lymphocyte Technology Testing Therapeutic Time Toxic effect Work base cell fixing clinically relevant companion diagnostics cytokine immunotoxicity improved instrument meetings member microchip oncology patient stratification personalized management programs response success tool treatment response tumor

项目摘要

Despite the demonstrated benefit of CD19 targeted CAR-T immunotherapeutics, two challenges remain to bringing the therapeutics to market. The first key challenge is to reproducibly manufacture the therapy so that the cells' cytokine mediated function acts in a predictably consistent fashion post manufacturing. The second key challenge is to manage the cell's adverse effect (immuno-toxicity), namely cytokine release syndrome (CRS) of IL-6 and other inflammatory cytokines, while also ensuring efficacy against the CD19 target. A more effective pre-infusion quality control test to ensure consistent and safe functioning would be not only address these two critical challenges, but help pharmaceutical firms satisfy FDA concerns in both of these areas. Such a test would have deep impact on ensuring the pharmas help these therapies to market, and in making sure that patients in need with incurable B-cell malignancies have access to these revolutionary therapies. A more reliable in vitro CAR-T functional test would allow physicians to remove or modify the inconsistent or unsafe cell therapies prior to injection, significantly reducing risk to the patient, and improving odds of therapeutic success. However, the current leading pre-infusion tests do not address the most important clinical requirements. The first requirement of such a test is to evaluate the CAR functional cell cytokine activation in the single-cell poly-functional cell subsets, a correlate of quality immune response, and also monitor for adverse functional reactions amongst these cell subsets. That requires up to 18 non-overlapping cytokines per cell. A second requirement is to measure the cells' true secretions in an “ex vivo” manner, rather than fixing the cells and manipulating their true function. IsoPlexis' microchip technology meets these two needs for the first time. It measures the range of efficacy polyfunctional cytokine markers (anti-tumor, stimulatory, chemotactic), while measuring the CRS related inflammatory and also regulatory cytokines from those subsets (up to 42 cytokines per cell). It does so in an ex vivo platform that not only measures true cell secretion to CD19 target, but can also interact with and respond to the target cell directly on device. IsoPlexis plans to use its core technology and learning to create an in vitro assessment test to measure for consistent range of function for in CAR-T cell therapies and to monitor for markers of safety and efficacy. We propose to do the following specific aims: (1) Develop a panel and reproducible assay for measurement of CD19+ CAR cells upon interaction with CD19 (immobilized) to ensure consistent function to CAR target. (2) Implement the assay above with transfected CAR-T donor samples, and demonstrate a clinically useful workflow. With the Phase I award support, we expect in the future to use it as a companion diagnostics tool to monitor the response of patients and ensure proper management of these personalized living drugs in every patient, justifying the broad impact of the proposed microdevice in immune-oncology.

尽管 CD19 靶向 CAR-T 免疫疗法已被证明具有益处，但仍面临两个挑战将疗法推向市场的第一个关键挑战是可重复地生产疗法，以便细胞的细胞因子介导的功能在制造后以可预测的一致方式发挥作用。关键的挑战是控制细胞的副作用（免疫毒性），即细胞因子释放综合征 (CRS) IL-6 和其他炎症细胞因子，同时还确保针对 CD19 A 靶标的功效。有效的输注前质量控制测试以确保一致和安全的功能不仅要解决这两个关键挑战，但可以帮助制药公司满足 FDA 在这两个领域的担忧。测试将对确保制药公司帮助这些疗法进入市场产生深远的影响，并确保患有无法治愈的 B 细胞恶性肿瘤的患者可以获得这些革命性的疗法。可靠的体外 CAR-T 功能测试将使医生能够消除或修改不一致或不安全的注射前进行细胞疗法，显着降低患者的风险，并提高治疗的几率然而，目前领先的输注前测试并未解决最重要的临床问题。此类测试的首要要求是评估 CAR 功能细胞细胞因子的激活。单细胞多功能细胞亚群，与质量免疫反应相关，并监测这些细胞亚群之间的不良功能反应需要多达 18 个非重叠细胞因子。第二个要求是以“离体”方式测量细胞的真实分泌物，而不是固定细胞。 IsoPlexis 的微芯片技术首先满足了这两个需求。它测量多功能细胞因子标记物的功效范围（抗肿瘤、刺激、趋化），同时测量这些子集（最多 42 个）中与 CRS 相关的炎症和调节细胞因子它是在离体平台上进行的，该平台不仅可以测量 CD19 靶标的真实细胞分泌，但也可以直接在设备上与目标细胞交互并做出响应 IsoPlexis 计划使用其核心。技术和学习创建体外评估测试，以测量体内功能的一致范围 CAR-T细胞疗法的安全性和有效性标记物的监测我们建议做以下具体工作。目标：(1) 开发一套可重复的检测方法，用于测量 CD19+ CAR 细胞与 CD19（固定化）以确保 CAR 靶标的功能一致 (2) 执行上述测定。转染 CAR-T 供体样本，并展示了临床上有用的工作流程，获得了一期奖。支持，我们希望将来将其用作伴随诊断工具来监测患者的反应并确保对每位患者进行这些个性化生活药物的适当管理，证明其广泛的影响是合理的所提出的免疫肿瘤学微型装置。