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

项目摘要 迫切需要新的诊断和预后生物标志物来改善癌症的临床结果。 循环外泌体正在成为用于非侵入性癌症诊断的“液体活检”的新范例， 监测.外泌体是由大多数细胞分泌的大小为30-150 nm的小膜囊泡， 越来越多的证据显示外来体的重要生物学作用和临床相关性。以各种 在癌症中，已经发现肿瘤来源的外泌体在人类生物流体如血液中积累， 富含一组来自原始细胞的生物分子，例如特定的蛋白质和RNA，其可以 构成“肿瘤标记”。然而，对外泌体的临床价值的评估在一定程度上受到限制， 这是由于分离和分析这种小的、动态的和分子多样性的囊泡的挑战。到 为了解决分析技术和精准医学方面的这一明显差距，我们在这里提出了一个 外泌体指纹纳米设备（ExoFIND）系统，提供前所未有的分析能力， 测量存在于循环中的肿瘤来源的外来体携带的囊内蛋白标记物。这个新 该技术将适用于检测来自细胞系、动物模型和人类生物流体的外泌体 评估外泌体蛋白，包括Hu抗原R（HuR），作为潜在的液体活检为基础的标记物， 癌症的诊断、预后和靶向治疗。将进行拟议的研究，以实现 4个具体目标：1）开发通用的、高分辨率的喷墨胶体打印技术， 3D纳米工程生物测定装置的可扩展制造; 2）建立3D纳米工程ExoFIND 肿瘤外泌体指纹综合分析系统; 3）对ExoFIND系统进行改造和验证 作为基于血液的检测平台，评估结直肠癌（CRC）衍生的外泌体作为生物标志物 肿瘤发生和进展; 4）调整和验证ExoFIND系统作为伴随检测，以监测 HuR靶向治疗的肿瘤反应、耐药性和预后。我们的研究取得了成功， 将为以下方面提供重要动力：（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，和（2）催化探索 外来体作为精确肿瘤学的液体活检的新范例。我们的首要目标是最终 为广泛的基础和翻译研究提供了一个变革性的平台技术， 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.