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

基本信息

批准号：
10633266
负责人：
JINA KO
金额：
$ 24.3万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
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/10633266
关键词：
Achievement Address Antibodies Biological Markers Biology Cancer Patient Cell Communication Cells Chemistry Clinical Collaborations Communication Data Detection Development Diagnostic Dimensions Disease Encapsulated Fine needle aspiration biopsy General Hospitals Generations Glioblastoma Goals Harvest Heterogeneity Hospitals Immune Immunology Immunotherapy Individual 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 Prediction of Response to Therapy Research Research Personnel Resources Sampling Signal Transduction Stains Systems Biology Technology Testing Therapeutic Treatment outcome Universities Vesicle Woman Work biological heterogeneity biological systems biomarker discovery blood-based biomarker cancer biomarkers cancer cell career cell fixing cell type clinical diagnostics digital disease diagnosis disease diagnostic disorder subtype exosome extracellular vesicles fluorescence imaging fluorophore forging improved liquid biopsy medical schools minimally invasive molecular diagnostics molecular imaging mouse model neoplastic cell neurogenetics neurosurgery next generation next generation sequencing novel particle responders and non-responders response tool translational medicine treatment response tumor tumor-immune system interactions

项目摘要

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.

该提案旨在开发一个超高灵敏度平台，可以绘制细胞和细胞外的图谱单颗粒水平的囊泡（EV）分子信息，并将其应用于肿瘤免疫分析用于免疫治疗监测的微环境（TIME）。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分析身体状态以进行疾病诊断。该提案假设单细胞-单EV图谱将为细胞间通讯提供新的分子信息以更好地了解病理发展和疾病诊断。本次活动的目标提案有三重：i) 开发单一 EV 蛋白测序技术来解决 EV 问题异质性并发现疾病生物标志物的罕见 EV 亚型，ii) 超快循环多重活细胞荧光成像可监测细胞表型的变化识别多种免疫细胞类型，以及 iii) 单细胞-单 EV 作图以发现新的免疫细胞类型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治疗效果。 Jina Ko（该项目的 PI）拥有微流控、分子生物学和机器学习方面的专业知识提案）开发了用于液体活检的新的微米和纳米技术，可以提取来自血液生物标志物（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颖的平台，作为该提案的基础工作。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速循环。通过这项工作，Jina 锻造了马萨诸塞州总医院和哈佛大学内外的研究合作维斯研究所和哈佛大学医学院。在这些成就的基础上，这项工作将由液滴微流体和物理学领域的世界专家团队执行 (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（Chiocca），他们将提供完整的对这项工作的支持。与这些导师合作将使她能够应对临床挑战问题并进一步发展她作为独立调查员的职业生涯，并有能力发展下一代医疗诊断。