Hydrogel microparticle technology for high-throughout screening of chimeric antigen receptor-T cells based on single cell effector function

基于单细胞效应功能的嵌合抗原受体T细胞高通量筛选水凝胶微粒技术

• 批准号: 10604170
• 负责人: Joseph de Rutte
• 金额: $ 75.48万
• 依托单位: PARTILLION BIOSCIENCE CORPORATION
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-06 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10604170
• 关键词: Acceleration; Adoption; Affinity; Antigen Presentation; Antigen Targeting; Antigens; Area; Binding; Biological; Biological Assay; CAR T cell therapy; CD19 Antigens; CD19 gene; Cell Separation; Cell Therapy; Cell physiology; Cell secretion; Cells; Colon Carcinoma; Development; Effector Cell; Engineering; Environment; Equipment; Event; Formulation; Foundations; Funding; Gene Targeting; Generations; Grant; Hematologic Neoplasms; Hydrogels; In Vitro; Individual; Jurkat Cells; Libraries; Life; Ligands; Link; Macrophage; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Medicine; Molecular; Natural Killer Cells; Outcome; Pathway Analysis; Pharmaceutical Preparations; Price; Procedures; Production; Property; Protein Secretion; Raji Cell; Signal Transduction; Solid Neoplasm; Sorting; Structure; System; T-Cell Receptor; T-Lymphocyte; Technology; Therapeutic; Tissue Engineering; Transcript; Transmembrane Domain; Variant; base; cancer immunotherapy; cell killing; cellular transduction; chimeric antigen receptor; chimeric antigen receptor T cells; cost; cytokine; design; engineered T cells; exhaustion; experimental study; extracellular; fighting; fluorescence activated cell sorter device; functional improvement; gene therapy; immunoengineering; in vivo; instrument; malignant breast neoplasm; manufacture; multiple omics; new technology; next generation; novel; novel strategies; particle; physical property; population based; receptor; response; screening; single cell sequencing; success; therapeutic development; timeline; tool; transcriptome; transcriptomics; tumor microenvironment; tumor-immune system interactions

ABSTRACT Engineered cell therapies are becoming a pillar of medicine, along with molecular and genetic interventions. In particular, chimeric antigen receptor (CAR)-T cell therapies have had a number of FDA approvals in the last 5 years targeting hematologic malignancies. To extend the success of these therapies, especially for the treatment of solid tumors, a more thorough understanding of how CAR structure is linked to CAR-T cell function is necessary. Notably, widespread tools to analyze and select single CAR-T cells from a large population based on functional properties, such as secreted products or cytolytic activity, are critically lacking. Single-cell functional assays can enable screening a library of CAR designs introduced into a pool of cells to identify rare functional cells and sort these cells to recover CAR designs associated with important effector functions. If conducted in high throughput, thousands of constructs can be screened with hundreds of individual events per construct to have robust statistical accuracy in the linkage between function and sequence. In addition, if single-cell functional information can be tied to transcriptomic information, pathway analysis associated with strong effector functions can be performed, identifying other gene targets that improve function, even in the presence of immunosuppressive, exhaustion-prone, or other microenvironments associated with solid tumors. Partillion’s nanovial technology provides a new approach to measure the function of single cells using widely available fluorescence activated cell sorters (FACS) and single-cell sequencing instruments, which we aim to apply to cell therapy discovery. Here, we propose to develop single-cell secretion and cell-killing assays compatible with the nanovials to introduce a new product that would enable scaled cell therapy discovery workflows from millions of cells. We aim to identify optimal nanovial formulations and procedures to measure both cytokine production and cytolytic functions from the same cells, and then link single-cell transcriptomic information with this functional readout. We also will engineer nanovials to better recapitulate the tumor microenvironment, acting as an artificial antigen-presenting target cell with combinations of antigen and immunosuppressive signals. The approach should be applicable broadly beyond CAR-T cell therapies to other chimeric receptors in natural killer cells, and macrophages, or in finding engineered T cell receptors. Ultimately, more access to sophisticated cell selection approaches can lead to therapies that are both lower in price and more effective as well as expanding the scope of applications to un-explored therapeutic areas.

抽象的 工程细胞疗法以及分子和遗传干预正在成为医学的支柱。 特别是，嵌合抗原受体（CAR）-T 细胞疗法在过去 5 年内获得了多项 FDA 批准 多年针对血液系统恶性肿瘤的治疗，以扩大这些疗法的成功，特别是治疗。 对于实体瘤，更深入地了解 CAR 结构如何与 CAR-T 细胞功能联系起来 值得注意的是，从大量群体中分析和选择单个 CAR-T 细胞的广泛工具是必要的。 严重缺乏对功能特性（例如分泌产物或细胞溶解活性）的研究。 分析可以筛选引入细胞池中的 CAR 设计库，以识别罕见的功能 细胞并对这些细胞进行分类以恢复与重要效应功能相关的 CAR 设计。 高通量，可以筛选数千个构建体，每个构建体有数百个单独的事件 此外，如果单细胞有功能，则在功能和序列之间的联系上具有稳健的统计准确性。 信息可以与转录组信息、与强效应子功能相关的通路分析联系起来 可以执行，识别其他改善功能的基因目标，即使存在 免疫抑制、易疲劳或其他与实体瘤相关的微环境。 纳米瓶技术提供了一种利用广泛可用的方法来测量单细胞功能的新方法 荧光激活细胞分选仪（FACS）和单细胞测序仪器，我们的目标是将其应用于细胞 在这里，我们建议开发与以下方法兼容的单细胞分泌和细胞杀伤测定法。 nanovials 推出了一种新产品，该产品将使数以百万计的细胞治疗发现工作流程规模化 我们的目标是确定最佳的纳米瓶配方和程序来测量细胞因子的产生和检测。 来自相同细胞的溶细胞功能，然后将单细胞转录组信息与该功能联系起来 我们还将设计纳米小瓶，以更好地再现肿瘤微环境，充当人工的角色。 结合抗原和免疫抑制信号的抗原呈递靶细胞。 应该广泛适用于 CAR-T 细胞疗法之外的自然杀伤细胞中的其他嵌合受体，并且 巨噬细胞，或寻找工程化 T 细胞受体，最终可以更容易地进行复杂的细胞选择。 方法可以带来价格更低、更有效以及扩大范围的治疗方法 未探索的治疗领域的应用。