Integrative Functional Profiling of Tumor-Derived Extracellular Vesicles

项目摘要 精准医疗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，克服耐药性， 复发为了应对这些挑战，对更好的生物标志物和测试存在关键的未满足的需求， 补充目前的准确诊断、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方式，因为它有望提供全球范围的 肿瘤动力学的观点细胞外囊泡（EV），包括外泌体，正在成为一种新的细胞外生物学的范例。 液体活检用于非侵入性癌症诊断和监测。外泌体是40-150 nm的膜囊泡 由大多数细胞分泌，并已被鉴定为细胞相互作用和信号传导的重要介质， 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， 外泌体的精确生物学功能，包括基质金属蛋白酶（MMPs）介导的 肿瘤微环境，其潜在的临床价值仍有待确定。这部分是由于 在分离和分析这些具有不同分子和功能的纳米囊泡方面的艰巨挑战 特性.在这里，我们假设循环外泌体的功能表型可以提供有效的 用于检测早期恶性肿瘤、监测肿瘤进展和转移以及评估治疗的生物标志物 乳腺癌的治疗为了验证这一假设，我们提出了先进的发展和验证的一个 纳米工程微流控生物传感系统，能够集成分析分子和功能 在一个简化的工作流程中获得外泌体的表型。本研究将通过三个具体目标来进行：1） 扩展MINDS战略，开发最佳的3D纳米工程一体化EV分子和活性 EV-MAP纳米芯片平台; 2）调整和优化EV-MAP技术用于监测肿瘤 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的潜在应用。这项新技术将 赋予上级分析能力以显著加速循环外来体的功能研究。 利用外泌体活性用于诊断、预后或治疗益处提出了一种范式转变 精准医疗的机制。虽然在这个项目中重点关注乳腺癌，但我们的研究最终将 为各种恶性肿瘤中广泛的生物活性外泌体的研究创造了一个变革性的工具， 可靠的非侵入性癌症液体活检。

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.