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: 10219990
• 负责人: Ajay Goel
• 金额: $ 51.45万
• 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/10219990
• 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; Structure; 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) 是癌症相关死亡的第二大原因 然而，与其他癌症不同，大多数早期结直肠癌可以通过手术治愈。 检测 CRC 并切除前驱病变，特别是晚期结直肠腺瘤 (A-CRA) 被认为是降低结直肠癌相关死亡率的最佳方法，然而，目前可用的筛查。 尽管结肠镜检查作为筛查工具很有效，但其侵入性和 费用常常阻碍人们遵循 CRC 筛查指南；并且基于粪便的检测不准确； 作为一种诊断方式仍然不够充分，越来越多的证据表明 microRNA 的失调。 (miRNA) 存在于所有人类癌症中，作为生物标志物，miRNA 比 mRNA 更具弹性，因为它们的弹性较小。 与相比，易于降解，并且即使在肿瘤形成的最早阶段也经常放松管制 此外，最近发现癌症会主动分泌小细胞外囊泡， 被称为“外泌体”的进入系统循环，为转化领域带来了额外的热情 尽管外泌体因其结构稳定性和稳定性而被认为是有前景的。 由于分子谱反映了其细胞来源，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) 缺乏分离和纯化的标准化方案；ii) 使用 用于生物标志物发现的细胞系样本，而非患者样本 iii) 缺乏分子分析研究； 对来自匹配组织和血浆样本的癌症来源的外泌体进行研究，以确定其癌症特异性； iv) 尽管人们认为外泌体 miRNA (exo-miRNA) 可能优于循环无细胞 miRNA （cf-miRNA），没有研究直接比较这两种类型，以支持或 否定这两种类型作为临床相关疾病生物标志物的优越性。在本提案中，我们将讨论这一点。 通过实现以下具体目标 1：优化患者来源的外泌体来解决这些问题。 分离，然后基于 RNA-Seq 发现循环无细胞 (cf-miRNA) 和外泌体 miRNA (exo- 从 A-CRA 患者、CRC 和个体采集的匹配组织和血浆样本中的 miRNA） 目标 2：开发循环无细胞和外泌体 miRNA 生物标志物组 将 A-CRA 和 CRC 患者与健康个体区分开来 目标 3：临床验证和表现。 在大型、独立的结直肠癌患者队列中评估优化的 cf-miRNA 和 exo-miRNA 瘤形成。 如果成功，该提案将提供急需的无细胞和外泌体的分子表征 miRNA 生物标志物作为液体活检生物标志物，可能将 CRC 的早期检测转变为稳健的、非 侵入性且廉价的临床检测。