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.

项目摘要/摘要 细胞外囊泡（EV）包括一组含有一个的分泌膜结构 多种生物分子货物，例如核酸，蛋白质和脂质。电动汽车是液体活检的理想选择 癌细胞通常比正常细胞更高度分泌癌细胞，可以在癌症的体液中检测到。 病人，保护其货物免受降解。因为EV货物通常反映了遗传和生物学 原籍单元的状态，EV货物的成分是癌症诊断的有希望的生物标志物 和复发。在这些成分中，miRNA作为潜在的生物标志物引起了很大的关注。 但是，几项研究表明，很大一部分电动汽车不包含明显的miRNA拷贝 数字。这些研究暗示了富含miRNA的EV的亚群的存在， 但是尚未确定这种亚群。这项研究的总体目标是开发方法 为了液体活检目的，富含miRNA的电动汽车的分离亚群。我们的初步研究 已经根据其表面蛋白表达确定了EV的几种不同的亚群，并暗示 表面蛋白库可以鉴定出富含miRNA的电动汽车。在这项研究中，我们首先将 评估人类癌细胞分泌并存在于三个主要患者的体液中的电动汽车 癌症的类型（卵巢，结直肠，肾脏），用于其表面定义的EV亚群的患病率 蛋白质库。其次，我们将确定EV的每个亚群中的总miRNA含量 由人类癌细胞分泌并存在于癌症患者体液中。第三，我们将评估检测 电动汽车中的miRNA通过从肿瘤的体液中免疫接种而分离出来的电动汽车中的miRNA 轴承小鼠和癌症患者。如果成功，我们的研究将为关键的知识差距提供答案 EV生物标志物领域，重要的是，开发了一种可以轻松隔离miRNA富含电动汽车的方法 用于临床实验室环境用于液体活检。