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

• 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
• 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

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有针对性的CAR-T免疫疗法表现出了表现的好处，但仍有两个挑战 将治疗剂推向市场。第一个关键挑战是可重复制造该疗法，以便 细胞的细胞因子介导的功能在制造后的时尚后的一致性时期起作用。第二个 关键挑战是管理细胞的不良影响（免疫毒性），即细胞因子释放综合征 IL-6和其他炎症细胞因子的（CRS），同时还确保针对CD19靶标的效率。更多 有效的注入前质量控制测试以确保一致且安全的运作不仅是地址 这两个关键的挑战，但有助于制药公司满足这两个领域的FDA关注。这样的 测试将对确保药物帮助这些疗法进行市场产生深远影响，并确保 有需要治疗的B细胞Malignancys有需要的患者可以使用这些革命性的疗法。更多 可靠的体外CAR-T功能测试将使医生可以去除或修改不一致或不安全 注射前的细胞疗法，大大降低患者的风险，并提高治疗的几率 成功。但是，当前领先的灌注前测试并未解决最重要的临床 要求。这种测试的第一个要求是评估CAR功能性细胞因子激活 单细胞多功能细胞子集，质量免疫反应的相关性，还监视 这些细胞子集之间的不良功能反应。最多需要18种非重叠的细胞因子 细胞。第二个要求是以“离体”方式测量细胞的真实分泌物，而不是修复 细胞并操纵其真实功能。 Isoplexis的微芯片技术满足了这两个需求的第一个需求 时间。它测量了效率多功能细胞因子标记（抗肿瘤，刺激，趋化性）的范围， 同时测量与CRS相关的炎症性以及来自这些子集的调节细胞因子（多达42个） 每个细胞的细胞因子）。它在离体平台中这样做，该平台不仅测量了CD19目标的真实细胞分泌， 但也可以与设备上的目标单元直接互动并响应。等路计划使用其核心 技术和学习以创建体外评估测试，以衡量一致的功能范围 CAR-T细胞疗法并监测安全性和效率的标记。我们建议做以下特定的 目的：（1）开发一个面板和可重复的测定法，以测量与CD19+ CAR细胞相互作用后 CD19（固定）以确保CAR目标的一致功能。 （2）用上述测定 转染CAR-T供体样品，并展示了临床上有用的工作流程。与第一阶段奖 支持，我们希望将来将其用作伴侣诊断工具来监测患者的反应 并确保在每个患者中适当管理这些个性化的生物药物，证明广泛的影响 在免疫肿瘤学中提出的微电位。