Ultrasensitive quantification of cytokine release from T cells

T 细胞释放细胞因子的超灵敏定量

• 批准号: 10662547
• 负责人: Ying Samuel Hu
• 金额: $ 38.84万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662547
• 关键词: Acute; Area; Biological Assay; Biological Models; Calcium Channel; Cell Communication; Cell membrane; Cells; Clinical Trials; Complex; Disease; Drug Targeting; Enzyme-Linked Immunosorbent Assay; Event; FDA approved; Flow Cytometry; Future; Immune; Impaired wound healing; Interleukin-2; Investigation; Ion Channel; Life; Masks; Membrane; Molecular; Phenotype; Process; Regulation; Research; Resolution; Signaling Protein; Site; T-Cell Activation; T-Lymphocyte; Techniques; Technology; Time; Vesicle; clinical translation; cytokine; cytokine release syndrome; extracellular vesicles; immune imaging; inhibitor; nanoscale; novel; programs; screening; tool

PROJECT SUMMARY Cytokines represent a broad range of small signaling proteins utilized by immune cells in cell-to-cell communications. Dysregulated cytokine release contributes to acute and long-term conditions, from life- threatening cytokine release syndrome to delayed wound healing. Targeting the cytokine release is an attractive strategy because it can regulate multiple cytokines involved in complex diseases. However, no FDA- approved drugs target this process, and related clinical trials remain scarce. The lack of clinical translation represents an urgent need to develop advanced technologies to better understand the cytokine release process at the molecular level. To date, existing assays for cytokine quantification, such as ELISA and flow cytometry, provide a low resolution that masks detailed mechanistic information in space and time. By capitalizing on the PI’s unique expertise in immune imaging, the R35 proposal will address this need by developing ultrasensitive cytokine quantification techniques using T cells and interleukin-2 as a model system. Cytokine quantification will be achieved in three specific areas: 1) at the resolution of single-vesicle fusion events with the plasma membrane, 2) at the nanoscale membrane release sites, and 3) in the membrane- enclosed form of extracellular vesicles (EVs). Enhanced mechanistic understanding will be obtained at the single-cell level regarding the temporal and directional profiles of cytokine release, proximity-based regulation by membrane calcium channels, and the dynamic distribution between soluble and EV-associated cytokines during T cell activation. Each of these areas will potentially enable compound screening targeting specific spatial and temporal phenotypes, investigations of membrane channel inhibitors, and targeting EVs for cytokine modulation. Future studies will be expanded to other essential cytokines from adaptive and innate immune cells. Ultrasensitive quantification of cytokines will enhance mechanistic understanding of the cytokine release in search of novel membrane targets to modulate the process.

项目摘要 细胞因子代表了免疫细胞在细胞间相互作用中利用的广泛的小信号蛋白。 通讯失调的细胞因子释放有助于急性和长期的条件，从生活- 威胁细胞因子释放综合征以延迟伤口愈合。靶向细胞因子释放是一种 因为它可以调节复杂疾病中涉及的多种细胞因子。然而，没有FDA- 针对这一过程的获批药物，相关的临床试验仍然很少。缺乏临床翻译 迫切需要开发先进的技术来更好地了解细胞因子的释放 在分子水平上的过程。迄今为止，用于细胞因子定量的现有测定法，例如ELISA和流式细胞术， 流式细胞术提供了在空间和时间上掩盖详细的机械信息的低分辨率。通过 利用PI在免疫成像方面的独特专长，R35提案将通过以下方式满足这一需求： 开发使用T细胞和白细胞介素-2作为模型系统的超灵敏细胞因子定量技术。 细胞因子定量将在三个特定领域实现：1）在单个囊泡融合的分辨率下 与质膜的事件，2）在纳米级膜释放位点，和3）在膜中- 封闭形式的细胞外囊泡（EV）。将获得更深入的机械理解， 关于细胞因子释放的时间和方向分布的单细胞水平，基于邻近的调节 通过膜钙通道，以及可溶性和EV相关细胞因子之间的动态分布 在T细胞活化过程中。这些领域中的每一个都有可能使化合物筛选靶向特定的 空间和时间表型，膜通道抑制剂的研究，以及针对 细胞因子调节未来的研究将扩大到其他必需的细胞因子，从适应性和先天性 免疫细胞。细胞因子的超灵敏定量将增强对细胞因子机制的理解 释放以寻找新的膜靶点来调节该过程。