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

基本信息

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
项目类别：
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

项目摘要

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.

摘要/摘要工程细胞疗法与分子（药物和蛋白质）一样已成为医学的基石然而，需要新的工具来选择、分析和设计这些干预措施。 “活体药物”在过去几年中，在使用工程免疫细胞方面取得了特别成功。治疗血液恶性肿瘤的基础疗法，包括 FDA 最近批准的两种嵌合抗原不幸的是，这一成功并未广泛转化为更多产品。优化这些疗法的挑战之一是，各种分子疗法。由于结构和功能密切相关，细胞疗法更难以进行功能设计，因为基于表面标记表达或靶抗原亲和力的传统细胞分类效果不佳事实上，最近的单细胞筛选与抗癌功能相关，例如细胞因子分泌和细胞杀伤。强调了从同一患者中分离出的 T 细胞具有惊人的高水平功能多样性并具有相同的表面标记组，只有一小部分高度活跃的细胞驱动反应免疫挑战。过去几年出现了各种单细胞功能分析平台，但它们的广泛应用由于检测通量低、成本高或需要熟练的操作员和技术人员，其采用受到了限制。需要昂贵的定制仪器来揭示这些联系。 T 细胞分子和功能特性与抗癌活性之间的关系，最终使我们建议开发一个新颖的“细胞疗法实验室”。 “粒子”平台，可以将单个 T 细胞快速分离成大小均匀的纳米液滴，每个液滴由具有结构化空腔的微粒（称为纳米瓶）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，不了解微流体。使用纳米瓶需要或其他专门技术，我们的目标集中在：（1）开发纳米瓶。 T 细胞附着的最佳粘合特性以及与各种 FACS 仪器的兼容性； (2) 基于白介素-2 (IL-2) 和干扰素-γ 对个体抗原特异性 T 细胞进行分类和表征 (IFN-γ) 的产生我们将检验 T 细胞根据 IL-2 和 IFN-γ 的产生进行分类的假设，如下所示：在纳米瓶中测量，将改善效应器功能。进入功能性免疫细胞选择的重大障碍，并推动下一代癌症免疫治疗设计。