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.

项目摘要/摘要 在接下来的五年中，纳米仪将发展为一项技术，该技术提供了第一个外观 进入单细胞分泌组；具有单分子灵敏度。这个新的发展 技术基于将单细胞封装技术与纳米孔的整合 - 基于单分子传感。释放到细胞外的分子的集合 单细胞的环境称为单细胞分泌组，可对 细胞寿命和细胞通信的复杂性。技术的关键特征 将在项目期结束时开发的，包括（1）一种大规模平行的方式 形成液滴粘附在载玻片上并包含一个单元格，（2）基于纳米孔的 测量来自单个细胞的分泌分子的方法。一旦纳米孔 开发了相关方法，我们旨在回答有关重要的四个关键问题 生物医学问题。首先，纳米孔将量化单细胞抗体的产生 从已从各种疾病状态的患者中分离出来的人类浆细胞 包括自动免疫性疾病。其次，B细胞和T细胞将被共孵育并刺激 用单个抗原诱导免疫反应。开始生成抗体的时间滞后 以及影响免疫反应时机的因素。第三， 将研究与单个癌细胞（或癌症囊泡）共孵育的嗜酸性粒细胞 提供对早期癌症识别机制的见解。最后，单细胞分泌组 指纹将用于访问区分干细胞谱系的能力 非破坏性方式。干细胞表征对于细胞疗法很重要（其中 作用机理是分泌的分子），并评估细胞状态的再生状态 药品。不用说，分泌组在正常中扮演的功能却却被掩盖了 和患病的细胞状态。纳米的应用（一种基于纳米孔的方法 Semalome Analysis）将为​​单细胞表征提供独特的新工具。