Lab on a particle technology for functional screening of therapeutic cells

用于治疗细胞功能筛选的粒子技术实验室

• 批准号: 10272940
• 负责人: Dino Di Carlo
• 金额: $ 26.32万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-09 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10272940
• 关键词: Adhesives; Adoption; Affinity; Alpha Particles; Antibodies; Antibody Formation; Antigen Targeting; Antigens; Autologous; Automobile Driving; Bar Codes; Biological; Biological Assay; Biomedical Engineering; CD28 gene; CD3 Antigens; Caliber; Capital; Cell Adhesion; Cell Survival; Cell Therapy; Cell surface; Cell-Matrix Junction; Cells; Cellular biology; Cellular immunotherapy; Chemistry; Chinese Hamster Ovary Cell; Classification; Clinical; Clone Cells; Custom; Defense Mechanisms; Development; Diffuse; Disease; Encapsulated; Engineering; Ensure; Enzyme-Linked Immunosorbent Assay; Equipment; Fluorescence; Fluorescence-Activated Cell Sorting; Generations; Genetic; Hematologic Neoplasms; Hydrogels; Immune; Immunologics; Immunotherapy; Individual; Interferon Type II; Interferons; Interleukin-2; Intervention; Investments; Knowledge; Link; Literature; Lysine; Malignant Neoplasms; Measures; Mediating; Medicine; Microfabrication; Microfluidics; Molecular; Mus; Outcome; Ovalbumin; Particle Size; Patients; Peptides; Pharmaceutical Preparations; Phenotype; Population; Procedures; Process; Production; Property; Proteins; Recovery; Relapse; Reporting; Research; Research Personnel; Signal Transduction; Solid Neoplasm; Sorting - Cell Movement; Specificity; Stains; Standardization; Structure; Surface; System; T-Cell Receptor; T-Lymphocyte; T-cell diversity; Techniques; Technology; Testing; Therapeutic; Time; Transgenic Organisms; Translating; Tumor Immunity; anti-cancer; anticancer activity; antigen-specific T cells; base; cancer cell; cancer immunotherapeutics; cancer therapy; cell killing; chimeric antigen receptor; chimeric antigen receptor T cells; cohort; cost; cytokine; cytotoxicity; design; differential expression; fluorescence activated cell sorter device; gene therapy; immunoengineering; improved; in vivo; instrument; instrumentation; nanoDroplet; nanosized; new technology; next generation; novel; particle; patient subsets; receptor; response; sample fixation; screening; success; tool; treatment optimization; tumor

SUMMARY/ABSTRACT Engineered cell therapies have become a cornerstone of medicine, along with molecular (drugs and proteins) and genetic (gene therapy) interventions. However, new tools are needed to select, analyze, and design these “living drugs”. In the last few years, there has been particular success in the use of engineered immune cell- based therapies in treating hematologic malignancies, including recent FDA approvals of two chimeric antigen receptor (CAR)-T-cell products. Unfortunately, this success has not translated broadly, for example, to more prevalent solid tumors. One of the challenges in optimizing these therapies is that, unlike molecular therapies in which structure and function are intimately linked, cellular therapies are more difficult to functionally design, as conventional classifications of cells based on surface marker expression or target antigen affinity are poorly correlated with anti-cancer functions, such as cytokine secretion and cell killing. In fact, recent single-cell screens have highlighted an astonishingly high level of functional diversity from T-cells isolated from the same patient and bearing the same panel of surface markers, with only a small highly active subset of cells driving responses to immunological challenge. Various single-cell functional profiling platforms have emerged over the past several years, but their widespread adoption has been limited due to low assay throughputs, high-costs, or the need for skilled operators and expensive customized instrumentation. Broadly accessible technologies are needed to uncover the links between T-cell molecular and functional properties and anti-cancer activity, and ultimately, to enable the production and selection of the most efficacious cell therapies. We propose the development of a novel “lab on a particle” platform, which allows the rapid isolation of individual T-cells into uniformly sized nano-droplets, each formed by a microparticle with a structured cavity (termed a nanovial). This approach will provide simultaneous measures of both cell surface and secreted proteins, and recover cells with desired phenotypes at high rates using standard fluorescence-activated cell sorting (FACS) machines. Importantly, no knowledge of microfluidics or other specialized techniques is required to use nanovials. Our aims focus on: (1) developing nanovials with optimal adhesive properties for T-cell attachment and compatibility with a broad range of FACS instruments; and (2) sorting and characterizing individual antigen-specific T-cells based on interleukin-2 (IL-2) and interferon-γ (IFN-γ) production. We will test the hypothesis that T-cells sorted based on production of IL-2 and IFN-γ, as measured in nanovials, will have improved effector function. Our new technology promises to remove a significant barrier to entry in functional immune cell selection, and drive next-generation cancer immunotherapeutic design.

总结/摘要 工程细胞疗法已经成为医学的基石，沿着分子（药物和蛋白质） 基因治疗（Genetic Therapy）然而，需要新的工具来选择，分析和设计这些 “活毒品”。在过去的几年里，在使用工程免疫细胞方面取得了特别的成功- 包括最近FDA批准的两种嵌合抗原 CAR-T细胞产物。不幸的是，这一成功并没有广泛地转化为，例如， 常见的实体瘤。优化这些疗法的挑战之一是，与分子疗法不同， 由于细胞的结构和功能紧密相连，因此细胞疗法更难以进行功能性设计， 基于表面标志物表达或靶抗原亲和力的常规细胞分类很差 与抗癌功能相关，如细胞因子分泌和细胞杀伤。事实上，最近的单细胞筛选 已经强调了从同一患者分离的T细胞的高度功能多样性 并带有相同的表面标记，只有一小部分高度活跃的细胞驱动反应， 免疫挑战。 在过去的几年里，已经出现了各种单细胞功能分析平台，但是它们的广泛应用并不局限于此。 由于低分析通量、高成本或需要熟练的操作员， 昂贵的定制仪器。需要广泛使用的技术来揭示这些联系 T细胞分子和功能特性与抗癌活性之间的关系，并最终使 生产和选择最有效的细胞疗法。我们建议开发一种新的“实验室， 一个“粒子”平台，它允许将单个T细胞快速分离成大小均匀的纳米液滴， 由具有结构化空腔的微粒（称为纳米管）形成。这种方法将同时提供 测量细胞表面和分泌蛋白，并以高速率回收具有所需表型的细胞 使用标准荧光激活细胞分选（FACS）机器。重要的是，没有微流体的知识 或其它专门技术来使用纳米管。我们的目标集中在：（1）开发具有 最佳的粘附特性，可用于T细胞粘附，并与各种FACS仪器兼容;以及 (2)基于白细胞介素-2（IL-2）和干扰素-γ分选和表征单个抗原特异性T细胞 (IFN-γ）生产。我们将检验T细胞根据IL-2和IFN-γ的产生进行分选的假设， 在纳米管中测量，将具有改善的效应器功能。我们的新技术有望消除 功能性免疫细胞选择的重要进入障碍，并推动下一代癌症 免疫设计