NanoSMS: single molecule secretome analysis for non-destructive cellular fingerprinting

NanoSMS：用于非破坏性细胞指纹分析的单分子分泌组分析

• 批准号: 10713935
• 负责人: Kevin Freedman
• 金额: $ 34.69万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713935
• 关键词: Antibodies; Antibody Formation; Antigens; Autoimmune Diseases; B-Lymphocytes; Biology; Cell Communication; Cell Lineage; Cell Therapy; Cell physiology; Cells; Collection; Complex; Development; Disease; Encapsulated; Environment; Fingerprint; Glass; Human; Immune response; Life; Malignant Neoplasms; Measurement; Measures; Methodology; Modernization; Patients; Plasma Cells; Play; Population; Proteins; RNA; Regenerative Medicine; Retinal blind spot; Role; Slide; T-Lymphocyte; Techniques; Technology; Time; Vesicle; cancer cell; eosinophil; extracellular; insight; macromolecule; nanopore; new technology; single molecule; stem cells; tool

Project Summary/Abstract Over the next 5 years, NanoSMS will be developed into a technology which offers the first look into the single cell secretome; with single molecule sensitivity. The development of this new technology is based on the integration of single cell encapsulation technologies with nanopore- based single molecule sensing. The collection of molecules released into the extracellular environment by single cells, termed the single cell secretome, offers a unique glimpse into the complexities of cellular life and cell-to-cell communication. The key features of the technology which will be developed at the end of the project period include (1) a massively parallel way to form droplets adhered onto a glass slide and containing a single cell, and (2) a nanopore-based approach for measuring secreted molecule originating from a single cell. Once the nanopores and related methodologies are developed, we aim to answer four key questions surrounding important biomedical issues. First, the nanopore will quantify single cell antibody production originating from human plasma cells which have been isolated from patients in various disease states including auto-immune disorders. Second, B cells and T cells will be co-incubated and stimulated with a single antigen to induce an immune response. The time lag to start generating antibodies as well as the factors which influence the timing of the immune response will be studied. Third, eosinophils co-incubated with a single cancer cell (or cancer-derived vesicles) will be investigated to provide insight into early cancer recognition mechanisms. Lastly, single cell secretome fingerprinting will be used to access the ability to discriminate between stem cell lineages in a non-destructive manner. Stem cell characterization is important for cell therapy (in which the mechanism of action is secreted molecules) as well as assessing cell state for regenerative medicine. Needless to say, the secretome plays a functional and yet obscured role in both normal and diseased cellular states. The application of NanoSMS (a nanopore-based approach towards secretome analysis) will provide a unique and new tool for single cell characterization.

项目总结/摘要 在接下来的5年里，NanoSMS将发展成为一种技术， 进入单细胞分泌组;具有单分子敏感性。开发这个新 该技术基于单细胞封装技术与纳米孔的集成， 基于单分子传感。释放到细胞外的分子的集合 环境的单细胞，称为单细胞分泌组，提供了一个独特的一瞥， 细胞生命和细胞间通讯的复杂性。该技术的主要特点 将在项目期结束时开发的项目包括（1）大规模并行方式， 形成粘附在载玻片上并含有单个细胞的液滴，和（2）基于纳米孔的 用于测量源自单细胞的分泌分子的方法。一旦纳米孔和 相关的方法论的发展，我们的目标是回答四个关键问题，围绕重要的 生物医学问题。首先，纳米孔将量化单细胞抗体的产生， 从各种疾病状态的患者中分离的人浆细胞 包括自身免疫紊乱其次，将B细胞和T细胞共孵育并刺激 用单一抗原来诱导免疫反应产生抗体的时间差 以及影响免疫反应时机的因素将被研究。第三、 将研究与单个癌细胞（或癌源性囊泡）共孵育的嗜酸性粒细胞 以提供对早期癌症识别机制的深入了解。最后，单细胞分泌组 指纹识别将用于获得区分干细胞谱系的能力， 非破坏性的方式。干细胞表征对于细胞治疗是重要的（其中， 作用机制是分泌的分子）以及评估细胞再生的状态。 药不用说，分泌体在正常的和非正常的细胞中起着功能性的，但还不清楚的作用。 和病态的细胞状态NanoSMS（一种基于纳米孔的方法， 分泌组分析）将为单细胞表征提供独特的新工具。