Mapping Single Extracellular Vesicles to Parent Cells for Immunotherapy Monitoring

将单个细胞外囊泡映射到亲本细胞以进行免疫治疗监测

• 批准号: 10569343
• 负责人: JINA KO
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10569343
• 关键词: Achievement; Address; Antibodies; Biological Markers; Biology; Cancer Patient; Cell Communication; Cells; Chemistry; Clinical; Collaborations; Communication; Data; Detection; Development; Diagnostic; Disease; Encapsulated; Fine needle aspiration biopsy; General Hospitals; Generations; Glioblastoma; Goals; Harvest; Heterogeneity; Hospitals; Immune; Immunology; Immunotherapy; Individual; Institutes; Machine Learning; Maps; Massachusetts; Measures; Mentors; Methods; Microfluidics; Molecular; Molecular Analysis; Molecular Biology; Molecular Profiling; Monitor; Mothers; Nanotechnology; Neoplasm Circulating Cells; Nucleotides; Oncology; Outcome; Parents; Pathologic; Patient-Focused Outcomes; Peptide Sequence Determination; Phase; Phenotype; Physics; Population; Research; Research Personnel; Resources; Sampling; Signal Transduction; Stains; Systems Biology; Technology; Testing; Therapeutic; Treatment outcome; Tumor-infiltrating immune cells; Universities; Vesicle; Woman; Work; biological heterogeneity; biological systems; biomarker discovery; blood-based biomarker; cancer biomarkers; cancer cell; career; cell type; clinical diagnostics; digital; disease diagnosis; disease diagnostic; disorder subtype; exosome; extracellular vesicles; fluorescence imaging; fluorophore; improved; liquid biopsy; medical schools; minimally invasive; molecular diagnostics; molecular imaging; mouse model; neoplastic cell; neurogenetics; neurosurgery; next generation; next generation sequencing; novel; particle; predicting response; responders and non-responders; response; tool; translational medicine; treatment response; tumor; tumor-immune system interactions

This proposal aims to develop an ultra-high sensitive platform that can map cell and extracellular vesicle (EV) molecular information at the single particle level, and apply it to profile tumor immune microenvironment (TIME) for immunotherapy monitoring. This technology can resolve heterogeneity of biological systems and has the potential to discover robust biomarkers that can accurately profile status of the body for disease diagnostics. This proposal hypothesizes that single cell-single EV mapping will provide new molecular information on cell-cell communication for better understanding of pathological development and disease diagnosis. The goals of this proposal are threefold: i) development of single EV protein sequencing technology to resolve EV heterogeneity and discover rare EV subtypes for disease biomarkers, ii) ultra-fast cycling for multiplexed live cell fluorescence imaging to monitor the changes of cellular phenotypes and identify multiple immune cell types, and iii) single cell-single EV mapping to discover new molecular information on cell-cell communication through vesicle secretion. This technology will allow repeat sampling and monitoring of TIME during the course of immunotherapy and provide guidance to achieve the best possible patient outcomes. With the expertise in microfluidics, molecular biology, and machine learning, Jina Ko (the PI of this proposal) has developed new micro- and nano-technologies for liquid biopsy that can extract multidimensional molecular data from blood-based biomarkers (e.g. circulating tumor cells, EV). She has extended her expertise to chemistry, droplet microfluidics, and translational medicine to develop novel platforms that will serve as a fundamental work to this proposal. She has recently developed i) ultra-high sensitive single EV profiling technology using droplet digital PCR and ii) ultra-fast cycling for multiplexed cellular fluorescence imaging. Through this work, Jina has forged research collaborations within and outside the Massachusetts General Hospital and Harvard Medical School at the Wyss Institute and Harvard University. Building upon these achievements, this work will be executed with a team of world experts in droplet microfluidics and physics (Weitz), molecular imaging and diagnostics (Weissleder), immunology (Pittet), exosome biology and neurogenetics (Breakefield), and neurosurgery and oncology (Chiocca) who will provide a full support on this work. Working with these mentors will allow her to tackle clinically challenging problems and further develop her career as an independent investigator with the ability to develop next generation medical diagnostics.

该建议旨在开发一个可以绘制细胞和细胞外的超高敏感平台 囊泡（EV）分子信息在单个颗粒水平上，并将其应用于剖面肿瘤免疫 免疫疗法监测的微环境（时间）。这项技术可以解决 生物系统的异质性，并有可能发现可靠的生物标志物 准确地介绍了身体的疾病诊断状态。该提议假设 单个单元格映射将提供有关细胞 - 细胞通信的新分子信息 为了更好地理解病理发展和疾病诊断。目标的目标 建议是三重：i）开发单eV蛋白测序技术来解决EV 异质性并发现疾病生物标志物的罕见EV亚型，ii）超快速循环 多路复用的活细胞荧光成像，以监测细胞表型的变化和 识别多种免疫细胞类型，以及iii）单细胞单个单独的EV映射以发现新的 通过囊泡分泌的细胞 - 细胞通信的分子信息。这项技术将 在免疫疗法过程中重复对时间进行抽样和监测，并提供 实现最佳患者结果的指导。 凭借微流体，分子生物学和机器学习方面的专业知识，Jina Ko（PI的PI 提案）为液体活检开发了新的微技术和纳米技术，可以提取 来自基于血液的生物标志物（例如循环肿瘤细胞，EV）的多维分子数据。 她将专业知识扩展到化学，液滴微流体和转化医学上 开发新型平台，将作为该提案的基本工作。她最近有 开发i）使用液滴数字PCR和II的超高敏感单EV分析技术） 超快速循环用于多路复用细胞荧光成像。通过这项工作，Jina锻造了 马萨诸塞州综合医院和哈佛大学内外的研究合作 Wyss研究所和哈佛大学的医学院。以这些成就为基础 这项工作将与液滴微流体和物理专家团队一起执行 （Weitz），分子成像和诊断（Weissleder），免疫学（Pittet），外泌体生物学 和神经遗传学（Breakefield）以及神经外科和肿瘤学（Ch​​iocca），他们将提供完整 支持这项工作。与这些导师合作将使她能够解决临床上的挑战 问题并进一步发展她作为独立调查员的职业，能够发展 下一代医学诊断。