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.

该提案旨在开发一种超高灵敏度的平台，可以映射细胞和细胞外 囊泡（EV）分子信息，并将其应用于肿瘤免疫谱 微环境（TIME）用于免疫治疗监测。这项技术可以解决 生物系统的异质性，并有可能发现强大的生物标志物， 准确地描绘出身体的状态，用于疾病诊断。该提案假设， 单细胞-单EV作图将为细胞间通讯提供新的分子信息 以便更好地了解病理发展和疾病诊断。这个的目标 建议有三个方面：i）开发单一EV蛋白测序技术以解决EV 异质性和发现疾病生物标志物罕见EV亚型，ii） 多重活细胞荧光成像以监测细胞表型的变化， 鉴定多种免疫细胞类型，和iii）单细胞-单EV作图以发现新的 通过囊泡分泌进行细胞间通讯的分子信息。这项技术将 允许在免疫治疗过程中重复采样和监测TIME，并提供 指导以实现最佳的患者结局。 凭借在微流体，分子生物学和机器学习方面的专业知识，Jina Ko（该项目的PI） 提案）已经开发了用于液体活检的新的微米和纳米技术， 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。 她将自己的专业知识扩展到化学，液滴微流体和转化医学， 开发新的平台，将作为这项建议的基础工作。她最近 开发了i）使用液滴数字PCR的超高灵敏度单EV分析技术和ii） 用于多重细胞荧光成像的超快循环。通过这项工作，吉娜锻造了 马萨诸塞州总医院和哈佛内外的研究合作 韦斯利学院和哈佛大学医学院。在这些成就的基础上， 这项工作将由液滴微流体学和物理学方面的世界级专家组成的小组来完成 （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（布雷克菲尔德），神经外科和肿瘤学（Chiocca）谁将提供一个完整的 支持这项工作。与这些导师一起工作将使她能够应对临床挑战 问题，并进一步发展她的职业生涯，作为一个独立的调查员，有能力发展 下一代医疗诊断