Exosomal Biomarkers for the Noninvasive Detection of Colorectal Cancer

用于无创检测结直肠癌的外泌体生物标志物

• 批准号: 10478874
• 负责人: Ajay Goel
• 金额: $ 50.83万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-20 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10478874
• 关键词: Address; Bioinformatics; Biological Assay; Biological Markers; Blood Circulation; CA-19-9 Antigen; Cancer Etiology; Cell Line; Cells; Cessation of life; Clinical; Colon; Colonoscopy; Colorectal Adenoma; Colorectal Cancer; Colorectal Neoplasms; Colorectal Polyp; Development; Diagnostic; Disease; Early Diagnosis; Evaluation; Excision; Generations; Human; Individual; Lesion; Malignant Neoplasms; Messenger RNA; Methodology; MicroRNAs; Modality; Molecular; Molecular Profiling; Mucous Membrane; Mutation; Nature; Neoplasms; Non-Invasive Cancer Detection; Operative Surgical Procedures; Patients; Perception; Performance; Plasma; Prevention; Protocols documentation; Reproducibility; Research; Research Personnel; Resources; Retrospective cohort; Sampling; Screening procedure; Small RNA; Solid; Specificity; Specimen; Standardization; Talents; Testing; Tissues; United States; Validation; base; biomarker discovery; biomarker panel; circulating microRNA; clinically relevant; cohort; colon cancer patients; colorectal cancer screening; diagnostic biomarker; diagnostic screening; early detection biomarkers; exosome; extracellular vesicles; genome-wide; improved; liquid biopsy; microRNA biomarkers; mortality; noninvasive diagnosis; prognostic; screening guidelines; transcriptome sequencing

PROJECT SUMMARY: Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in the United States. Yet, unlike other cancers, the majority of early stage CRCs are surgically curable. Therefore, early detection of CRCs and removal of precursor lesions, particularly advanced colorectal adenomas (A-CRA), is considered as the best approach in reducing CRC-associated mortality. However, current available screening procedures are impractical. In spite of the efficacy of colonoscopy as a screening tool, its invasive nature and expense often dissuade individuals to follow CRC screening guidelines; and inaccuracy of fecal-based tests remains insufficient as a diagnostic modality. Accumulating evidence indicates that dysregulation of microRNAs (miRNAs) occurs in all human cancers. As biomarkers, miRNAs are more resilient than mRNAs as they are less prone to degradation, and are frequently deregulated even in the earliest stages of neoplasia compared to genetic alterations. Furthermore, the recent discovery that cancers actively excrete small extracellular vesicles, called “exosomes”, into systemic circulation, has brought additional enthusiasm to the field of translational biomarker research. Even though exosomes are considered promising due to their structural stability and molecular profiles reflecting their cell-of-origin, utilization of exosomes in biomarker research has been hampered due to multiple reasons, including: i) lack of standardized protocols for their isolation and purification; ii) use of cell line-derived, not patient-derived specimens for biomarker discovery; iii) lack of molecular profiling studies on cancer-derived exosomes from matched tissues and plasma specimens to establish their cancer specificity; iv) albeit the perception that exosomal-miRNAs (exo-miRNAs) may be superior to circulating cell-free miRNAs (cf-miRNAs), no studies have undertaken an effort to directly compare these two types, in order to support or negate the superiority of either type, as clinically-relevant disease biomarkers. In this proposal, we will address these concerns by undertaking the following Specific Aims. Aim 1: Optimization of patient-derived exosome isolation, followed by RNA-Seq based discovery of circulating cell-free (cf-miRNAs) and exosomal miRNAs (exo- miRNAs) in matched tissue and plasma specimens collected from patients with A-CRAs, CRCs and individuals with normal colon. Aim 2: Development of circulating cell-free and exosomal miRNA biomarker panels that distinguish patients with A-CRAs and CRCs from healthy individuals. Aim 3: Clinical validation and performance evaluation of optimized cf-miRNAs and exo-miRNAs in large, independent patient cohorts with colorectal neoplasia. If successful, this proposal will provide much needed molecular characterization of cell-free and exosomal miRNA biomarkers as liquid biopsy biomarkers, which may transform early-detection of CRC into a robust, non- invasive, and inexpensive clinical assay.

项目摘要：结直肠癌（CRC）是与癌症相关死亡的第二大原因 美国。然而，与其他癌症不同，大多数早期CRC在手术上都是可以治愈的。因此，早 检测CRC和去除前体病变，特别是晚期大肠腺瘤（A-CRA），是 被认为是降低CRC相关死亡率的最佳方法。但是，目前可用的筛选 程序是不切实际的。尽管结肠镜检查作为筛查工具的效率，但其侵入性和 费用通常会劝阻个人遵循CRC筛查指南；和基于粪便的测试不准确 作为诊断方式仍然不足。积累的证据表明microRNA的失调 （miRNA）发生在所有人类癌症中。作为生物标志物，miRNA比mRNA更具耐药性，因为它们较少 容易降解，即使在肿瘤的最早阶段，也经常受到管制。 遗传改变。此外，最近的发现是癌症积极地出色的小细胞外蔬菜， 被称为“外泌体”进入系统圈子，为翻译领域带来了更多的热情 生物标志物研究。即使外泌体由于其结构稳定性而被视为承诺和 反映其原始细胞的分子特征，外泌体在生物标志物研究中的利用受到了阻碍 由于多种原因，包括：i）缺乏标准化协议来隔离和纯化； ii）使用 细胞系衍生的，而不是生物标志物发现的患者衍生标本； iii）缺乏分子分析研究 从匹配的组织和血浆标本的癌症衍生的外泌体上，以建立其癌症特异性； iv）尽管认为外泌体miRNA（exo-miRNA）可能优于无细胞的miRNA （CF-MIRNA），没有进行任何研究直接比较这两种类型，以支持或 否定两种类型的优越性，作为临床上与疾病的生物标志物。在此提案中，我们将解决 通过实现以下特定目标来关注这些问题。目标1：优化患者衍生的外泌体 分离，然后是基于RNA-seq的发现无细胞的发现（CF-MIRNA）和外泌体miRNA（exo- miRNA）在从A-CRA，CRC和个体的患者中收集的匹配组织和血浆标本中 与正常结肠。目标2：开发无细胞和外泌体miRNA生物标志物面板 将A-CRA和CRC的患者与健康个体区分开。目标3：临床验证和性能 在大型，独立的患者队列中评估优化的CF-MIRNA和EXO-MIRNA 肿瘤。 如果成功，该提案将提供急需的分子表征，无细胞和外泌体表征 miRNA生物标志物作为液体活检生物标志物，可以将CRC的早期检测转化为可靠的非 - 非 - 侵入性和廉价的临床测定法。