Non-coding RNAs in serrated polyps identify patients with high colon cancer risk

锯齿状息肉中的非编码 RNA 可识别结肠癌高风险患者

• 批准号: 8969958
• 负责人: Don A Delker
• 金额: $ 20.51万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-07 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8969958
• 关键词: Accounting; Address; Adenomatous Polyps; Affect; Age; Benign; Biological; Biopsy Specimen; Blood; Caliber; Chemoprevention; Classification; Clinical; Code; Colon; Colon Carcinoma; Colonic Polyps; Colonoscopy; Data; Development; Diagnostic; Disease; Frequencies; Gene Expression; Gene Expression Profile; Genes; Genetic Predisposition to Disease; Genetic Transcription; Goals; Health; Hyperplasia; Hyperplastic Polyp; Large Intestine; Left; Length; Lesion; Messenger RNA; Metastatic to; MicroRNAs; Molecular; Outcome Study; Pathway interactions; Patient risk; Patients; Phenotype; Plasmids; Polyps; Procedures; Process; Proteins; Publishing; RNA; RNA Decay; RNA Sequences; Refractory; Risk; Sampling; Screening for cancer; Serum; Sigmoid colon; Site; Site-Directed Mutagenesis; Small RNA; Specimen; Syndrome; Technology; Testing; Tissues; Transfection; United States; Untranslated RNA; Untranslated Regions; Validation; Western Blotting; base; cancer risk; cell type; cohort; design; differential expression; high risk; improved; innovation; molecular marker; next generation sequencing; polyposis; screening; transcriptome sequencing; tumor

DESCRIPTION (provided by applicant): Serrated colon polyps are present in = 20% of patients undergoing routine screening colonoscopy. Serrated polyps are divided into sessile serrated and hyperplastic polyps and are often difficult to differentiate by endoscopic and histological examination. Although serrated polyps were previously considered harmless, recent studies provide evidence that sessile serrated polyps account for 20-30% of all colon cancers. We propose a multilateral approach to identify molecular diagnostic markers for sessile serrated polyps that are predictive of increased colon cancer risk. We will use the most innovative next generation sequencing technology to define the complete small RNA (< 200 nt) transcriptome in both serum and biopsy specimens from 20 patients with an exaggerated phenotype of sessile serrated polyps, known as serrated polyposis syndrome (SPS). The syndrome is defined as patients with = 5 sessile serrated polyps proximal to the sigmoid colon, two of which are > 1cm diameter, or patients with > 20 serrated polyps at any site in the large bowel. The age of presentation (often < 40 yrs) and multiplicity of lesions suggests a genetic predisposition, but it basis remains unknown and familial occurrence is unusual. To achieve this goal we have developed one of the largest cohorts of patients with the serrated polyposis syndrome in the world. We will also define the small RNA transcriptome in 20 patients with sporadic sessile serrated, hyperplastic and adenomatous polyps to identify a unique panel of microRNAs (miRNAs) specific to sessile serrated polyps. The analysis of small RNAs will aid in differentiating between polyp types similar to previously published tumor classification studies using miRNA panels. We have already collected 10-20 colon and 5 serum samples in each cohort and will collect another 10 colon and 15 serum samples in each cohort during the first year of study. Based on previous power calculations we are confident 20 patients in each cohort will be adequate to define differential expression of miRNAs across polyp types. We will use leave-one-out cross-validation to develop a two-step procedure for defining small RNA gene signatures in patients with sessile serrated polyps and patients with traditional hyperplastic polyps. We will validate ten miRNAs uniquely expressed in patients with sessile serrated polyps by qPCR in tissue and serum samples from patients in each cohort. We will identify and validate select miRNA targets using transient co-transfection of mRNA 3' UTRs and miRNA expression plasmids, site-directed mutagenesis of mRNA 3' UTRs and western blotting of mRNA products. Our hypothesis is that this approach will identify molecular markers that accurately differentiate sessile serrated polyps, in both the serrated polyposis syndrome and sporadic patients, from other polyps and provide a non-invasive serum test for identifying patients with an increased risk for colon cancer. The objective is to stratify patient's risk for colon cancer to guide the frequeny of cancer screening tests and aid the design of chemoprevention trials.

描述（由申请人提供）：存在常规结肠镜检查的患者中有20％的锯齿状结肠息肉。锯齿状的息肉分为无缝线和增生的息肉，通常很难通过内窥镜检查和组织学检查来区分。尽管锯齿状的息肉以前被认为是无害的，但最近的研究提供了证据表明，无链锯齿状的息肉占所有结肠癌的20-30％。我们提出了一种多边方法，以鉴定可预测结肠癌风险增加的无锯齿状息肉的分子诊断标记。我们将使用最具创新性的下一代测序技术来定义来自20名患有锯齿状锯齿状息肉表型的血清和活检标本中完整的小RNA（<200 nt）转录组，称为锯齿状息肉病综合征（SPS）。该综合征的定义为靠近乙状结肠的5个无锯齿状息肉的患者，其中两个直径> 1厘米，或大肠中任何部位的锯齿状息肉> 20个锯齿状息肉。表现的年龄（通常<40岁）和病变的多重性表明遗传易感性，但基础仍然未知，家族发生的情况是不寻常的。为了实现这一目标，我们开发了世界上最大的患有锯齿状息肉病综合征的患者之一。我们还将在20例偶发的锯齿状，增生和腺瘤息肉的患者中定义小的RNA转录组，以鉴定针对无孔锯齿状息肉的独特小组。小型RNA的分析将有助于区分使用miRNA面板类似于先前发表的肿瘤分类研究的息肉类型。我们已经在每个队列中收集了10-20个结肠和5个血清样品，在研究的第一年，每个队列中将再收集10个结肠和15个血清样品。根据先前的功率计算，我们对每个队列中的20名患者充满信心，足以定义跨类型的miRNA的差异表达。我们将使用剩余的交叉验证来开发两步程序，以定义有锯齿状息肉和传统增生性息肉患者的患者中的小RNA基因特征。我们将通过qPCR在每个队列中的患者的QPCR和血清样本中验证十种在静脉锯齿状息肉的患者中唯一表达的miRNA。我们将使用mRNA 3'UTR和miRNA表达质粒，mRNA 3'UTR的位置定向诱变以及mRNA产物的蛋白质印迹，使用mRNA 3'UTR和miRNA表达质粒的瞬时共染料来识别和验证选择的miRNA靶标。我们的假设是，这种方法将确定在锯齿状的息肉病综合征和散发性患者中，可以准确地分化无链锯齿状息肉，并与其他息肉相比，并提供非侵入性血清测试，以鉴定患有结肠癌风险增加的患者。目的是分层患者发生结肠癌的风险，以指导癌症筛查测试的频率并有助于化学预防试验的设计。