Integrative Functional Profiling of Tumor-Derived Extracellular Vesicles

项目总结 精准医学的临床实施面临着精准疾病分层和 分期，确定最佳治疗，监测治疗反应，并克服耐药性和 旧病复发。为了应对这些挑战，对更好的生物标记物和测试的迫切需求尚未得到满足 补充现有的准确诊断、预后和监测治疗反应的方法。液体 活检为精确肿瘤学提供了一种创新的非侵入性检查方式，因为它有望为全球 肿瘤动力学的观点。胞外囊泡(EV)，包括外周小体，正在作为一种新的 用于非侵入性癌症诊断和监测的液体活检。外切体是40-150 nm的膜小泡 由大多数细胞分泌，已被确认为细胞相互作用和信号转导的基本介质 促进肿瘤转移、耐药和复发。尽管这些发现具有潜在的临床影响， 外切体的精确生物学功能，包括基质金属蛋白酶(MMPs)介导的调控 肿瘤的微环境及其潜在的临床价值仍有待确定。这部分归因于 在分离和分析这些具有不同分子和功能的纳米囊方面面临的艰巨挑战 属性。这里我们假设循环外切体的功能表型可以提供有效的 检测早期恶性肿瘤、监测肿瘤进展和转移以及评估治疗的生物标志物 乳腺癌患者的反应。为了验证这一假设，我们建议对 可进行分子和功能综合分析的纳米工程微流控生物传感系统 一个简化的工作流程中的外切体表型。这项研究将由三个具体目标进行：1) 拓展思维策略，开发优化的3D纳米工程一体化电动汽车分子和活性 EV-MAP纳米芯片平台；2)将EV-MAP技术应用于肿瘤监测 使用小鼠模型评估和验证EV-MAP技术 对乳腺癌患者的临床诊断和分类具有潜在的应用价值。这项新技术将 提供卓越的分析能力，大大加快循环外切体的功能研究。 利用外切体活动进行诊断、预测或治疗是一种范式的转变 精准医疗机构。虽然在这个项目中专注于乳腺癌，但我们的研究最终将 创建一种变革性的工具，用于研究各种恶性肿瘤中的广泛生物活性外切体 可靠的癌症无创液体活检。

PROJECT SUMMARY Clinical implementation of Precision Medicine faces major challenges in precision disease stratification and staging, determining optimal treatment, monitoring therapy response, and overcoming drug resistance and relapse. To address these challenges, there is a critical unmet need for better biomarkers and tests that complement current methods for accurate diagnosis, prognosis and monitoring of response to treatments. Liquid biopsy presents an innovative non-invasive modality for precision oncology as it promises to provide a global view of tumor dynamics. Extracellular vesicles (EVs), including exosomes, are emerging as a new paradigm of liquid biopsy for non-invasive cancer diagnosis and monitoring. Exosomes are 40-150 nm membrane vesicles secreted by most cells and have been identified as essential mediators of cell interactions and signaling that promote tumor metastasis, drug resistance, and relapse. Despite the potential clinical impact of these findings, precise biological functions of exosomes, including matrix metalloproteinases (MMPs)-mediated modulation of tumor microenvironments, and their potential clinical value remain yet to be determined. This is due in part to the daunting challenges in isolation and analysis of these nanovesicles with diverse molecular and functional properties. Here we hypothesize that functional phenotypes of circulating exosomes can provide potent biomarkers for detecting early malignancy, monitoring tumor progression and metastasis, and assessing therapy response in breast cancer. To test this hypothesis, we propose the advanced development and validation of a nano-engineered microfluidic biosensing system capable of integrative analysis of both molecular and functional phenotypes of exosomes in one streamlined workflow. The research will be performed by three specific aims: 1) Expand the MINDS strategy to develop an optimal 3D nano-engineered integrative EV molecular and activity profiling (EV-MAP) nanochip platform; 2) Adapt and optimize the EV-MAP technology for monitoring tumor burden and therapy response using mouse models; and 3) Evaluate and validate the EV-MAP technology for potential applications to clinical diagnosis and classification of breast cancer patients. The new technology will confer superior analytical capabilities to substantially accelerate the functional studies of circulating exosomes. Harnessing exosome activities for diagnostic, prognostic or therapeutic benefit presents a paradigm-shifting mechanism for precision medicine. While focused on breast cancer in this project, our research will ultimately create a transformative tool for studies of a wide range of bioactive exosomes in various malignancies to develop reliable non-invasive liquid biopsy of cancer.