Nano-Engineered Lab-on-a-Chip for Assessing HuR-Regulated Exosomes for Cancer Monitoring and Targeted Therapy

项目总结 迫切需要新的诊断和预后生物标记物来改善癌症的临床结果。 循环外切体正在成为一种新的非侵入性癌症诊断和治疗的“液体活检”范例。 监控。外体是大多数细胞分泌的30-150纳米大小的膜小泡，以及 越来越多的证据表明，外切体具有重要的生物学作用和临床意义。在各种情况下 癌症，肿瘤衍生的外切体已被发现积聚在人体体液中，如血液，以及 富含来自起源细胞的一组生物分子，例如特定的蛋白质和RNA，这可能 构成一种“肿瘤征象”。然而，对外切体的临床价值的评估在一定程度上受到了限制。 由于在分离和分析这种小的、动态的和分子多样化的囊泡方面存在挑战。至 解决分析技术和精确医学方面的这一明显差距，在这里，我们建议 Exosome指纹纳米设备(ExoFIND)系统，提供前所未有的分析能力 测量循环中存在的肿瘤来源的外体携带的囊内蛋白标记物。这是一项新的 这项技术将被用于检测来自细胞系、动物模型和人体体液的外体。 评估外体蛋白，包括HU抗原R(HUR)，作为早期液体活检的潜在标记物 癌症的诊断、预后和靶向治疗。将进行拟议的研究，以实现 四个具体目标：1)开发一种通用、高分辨率喷墨胶体打印技术，用于标准化 和可扩展的3D纳米工程生物检测装置的制造；2)建立3D纳米工程ExoFIND 肿瘤胞外指纹综合分析系统；3)适配和验证ExoFIND系统 作为一个基于血液的检测平台，评估结直肠癌(CRC)来源的外切体作为生物标志物 肿瘤的发生和发展；4)采用和验证ExoFIND系统作为监测的配套试验 HUR靶向治疗的肿瘤反应、耐药性和预后。成功开展，我们的研究 将为以下方面提供重要推动：(1)开发具有变革性和 可重复的分析能力和最低的样品要求，以及(2)催化探索 Exosome作为精确肿瘤学液体活检的新范例。我们的首要目标是最终 为广泛的基础和翻译调查提供变革性的平台技术 恶性肿瘤和疾病。

PROJECT SUMMARY Novel diagnostic and prognostic biomarkers are urgently needed to improve clinical outcomes of cancer. Circulating exosomes are emerging as a new paradigm of “liquid biopsy” for non-invasive cancer diagnosis and monitoring. Exosomes are small membrane vesicles of 30-150 nm in size that are secreted by most cells, and growing evidence has shown important biological roles and clinical relevance of exosomes. In various cancers, tumor-derived exosomes have been found to be accumulated in human biofluids, such as blood, and enriched with a set of biomolecules from the cells of origin, such as specific proteins and RNAs, which may constitute a “tumor signature”. However, the evaluation of clinical value of exosomes has been limited in part due to challenges in isolation and analysis of such small, dynamic and molecularly diverse vesicles. To address this obvious gap in both analytical technologies and precision medicine, here we propose to an Exosome Fingerprinting Nano-Device (ExoFIND) system that offers unprecedented analytical capabilities for measuring intravesicular protein markers carried by tumor-derived exosomes present in circulation. This new technology will be adapted to examining exosomes derived from cell lines, animal models and human biofluids to assess exosomal proteins, including Hu antigen R (HuR), as potential liquid biopsy-based markers for early diagnosis, prognosis, and targeted therapy of cancer. The proposed studies will be carried out to accomplish four Specific Aims: 1) Develop a general, high-resolution inkjet colloidal printing technology for standardized and scalable fabrication of 3D nanoengineered bioassay devices; 2) Establish a 3D nano-engineered ExoFIND system for integrative analysis of exosomal fingerprints of tumors; 3) Adapt and validate the ExoFIND system as a blood-based detection platform to assess colorectal cancer (CRC)-derived exosomes as biomarkers for tumor initiation and progression; 4) Adapt and validate the ExoFIND system as a companion assay to monitor tumor response, drug resistance and prognosis of HuR-targeted therapy. Successfully carried out, our study will provide important impetus to: (1) develop next-generation technologies with transformative and reproducible analytical capabilities and minimal sample requirement, and (2) catalyze the exploration of exosomes as a novel paradigm of liquid biopsy for precision oncology. Our overarching goal is to ultimately provide a transformative platform technology for basic and translation investigation of a wide range of malignancies and diseases.