Isolation of miRNA-rich extracellular vesicles for liquid biopsy

分离富含 miRNA 的细胞外囊泡用于液体活检

• 批准号: 10431224
• 负责人: Honami Naora
• 金额: $ 18.93万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431224
• 关键词: Address; Antibodies; Area; Attention; Biogenesis; Biological; Biological Markers; Blood Circulation; Body Fluids; CD81 gene; Cancer Patient; Cells; Clinic; Clinical; Collaborations; Colorectal; Colorectal Cancer; Data Set; Detection; Disease; Electron Microscopy; Equipment; Flow Cytometry; Fluorescence; Genetic; Goals; Heterogeneity; Human; Kidney; Knowledge; Label; Laboratories; Lipids; Malignant neoplasm of ovary; Membrane Proteins; Methodology; Methods; MicroRNAs; Microfluidics; Microscopy; Mus; Normal Cell; Nucleic Acids; Ovarian; Pathway interactions; Patients; Plasma; Positioning Attribute; Prevalence; Prognosis; Proteins; RNA; Recurrence; Renal carcinoma; Resolution; Site; Structure; Surface; Translations; Tumor-Derived; Vesicle; Work; Xenograft Model; base; biomarker development; body volume; cancer biomarkers; cancer cell; cancer diagnosis; cancer type; clinically relevant; detection sensitivity; extracellular vesicles; innovation; interest; liquid biopsy; nanoparticle; novel strategies; overexpression; potential biomarker; protein expression; response; tumor

PROJECT SUMMARY/ABSTRACT Extracellular vesicles (EVs) comprise a heterogeneous group of secreted membranous structures that contain a variety of biomolecular cargo such as nucleic acids, proteins and lipids. EVs are ideal for liquid biopsy because EVs are often more highly secreted by cancer cells than by normal cells, can be detected in body fluids of cancer patients, and protect their cargo from degradation. Because EV cargo often reflects the genetic and biological status of the cell of origin, constituents of EV cargo are promising biomarkers for cancer diagnosis, prognosis and recurrence. Of these constituents, miRNAs have attracted substantial attention as potential biomarkers. However, several studies indicate that a substantial proportion of EVs do not contain significant miRNA copy numbers. These studies have implicated the existence of a subpopulation(s) of EVs that is enriched in miRNAs, but this subpopulation has not been identified. The overarching goal of this study is to develop approaches to isolate subpopulations of EVs that are enriched in miRNAs for liquid biopsy purposes. Our preliminary studies have identified several distinct subpopulations of EVs based on their surface protein expression, and implicate that the surface protein repertoire could identify EVs that are enriched in miRNAs. In this study, we will firstly evaluate EVs that are secreted by human cancer cells and present in body fluids of patients with three major types of cancers (ovarian, colorectal, renal) for the prevalence of EV subpopulations defined by their surface protein repertoire. Secondly, we will determine the total miRNA content in each subpopulation of EVs that are secreted by human cancer cells and present in cancer patient body fluids. Thirdly, we will evaluate the detection of miRNAs in EVs that are isolated by immunocapture of different EV surface proteins from body fluids of tumor- bearing mice and cancer patients. If successful, our study will provide answers to critical gaps-in-knowledge in the EV biomarker field and, importantly, develop an approach for isolating miRNA-rich EVs that can be readily used in a clinical laboratory setting for liquid biopsy purposes.

项目概要/摘要 细胞外囊泡（EV）由一组异质的分泌膜结构组成，其中含有 各种生物分子货物，例如核酸、蛋白质和脂质。 EV 是液体活检的理想选择，因为 癌细胞分泌的 EV 通常比正常细胞分泌的更多，可以在癌症的体液中检测到 患者，并保护他们的货物免遭降解。因为电动汽车货物往往反映了遗传和生物特征 起源细胞的状态、EV货物的成分是用于癌症诊断、预后的有前途的生物标志物 和复发。在这些成分中，miRNA 作为潜在的生物标志物引起了广泛的关注。 然而，多项研究表明，相当大比例的 EV 不包含显着的 miRNA 拷贝 数字。这些研究表明存在富含 miRNA 的 EV 亚群， 但这个亚群尚未被识别。本研究的总体目标是开发方法 分离富含 miRNA 的 EV 亚群，用于液体活检目的。我们的初步研究 根据 EV 的表面蛋白表达，确定了几个不同的 EV 亚群，并暗示 表面蛋白库可以识别富含 miRNA 的 EV。在本研究中，我们首先将 评估由人类癌细胞分泌并存在于三种主要癌症患者体液中的 EV 癌症类型（卵巢癌、结直肠癌、肾癌）的 EV 亚群患病率（由其表面定义） 蛋白质库。其次，我们将确定每个 EV 亚群中的总 miRNA 含量 由人类癌细胞分泌并存在于癌症患者的体液中。第三，我们将评估检测结果 EV 中的 miRNA 是通过从肿瘤体液中免疫捕获不同 EV 表面蛋白来分离的 携带小鼠和癌症患者。如果成功，我们的研究将为以下领域的关键知识空白提供答案： EV 生物标志物领域，重要的是，开发一种分离富含 miRNA 的 EV 的方法，该方法可以很容易地 在临床实验室环境中用于液体活检目的。