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

基本信息

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
项目类别：
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

项目摘要

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）：在接受常规结肠镜检查的患者中，有 20% 存在锯齿状结肠息肉。锯齿状息肉分为无蒂锯齿状息肉和增生性息肉，通过内镜和组织学检查往往难以区分。尽管锯齿状息肉以前被认为是无害的，但最近的研究提供的证据表明，无蒂锯齿状息肉占所有结肠癌的 20-30%。我们提出了一种多边方法来识别无蒂锯齿状息肉的分子诊断标记物，这些标记物可以预测结肠癌风险的增加。我们将使用最具创新性的下一代测序技术来定义 20 名患有夸张表型无蒂锯齿状息肉（称为锯齿状息肉病综合征 (SPS)）患者的血清和活检标本中的完整小 RNA (< 200 nt) 转录组。该综合征的定义是乙状结肠近端有≥ 5 个无蒂锯齿状息肉，其中两个直径 > 1 厘米，或在大肠任何部位有 > 20 个锯齿状息肉的患者。发病年龄（通常< 40岁）和病变的多样性提示有遗传倾向，但其基础仍然未知，并且家族性发生并不常见。为了实现这一目标，我们开发了世界上最大的锯齿状息肉病综合征患者队列之一。我们还将定义 20 名散发性无蒂锯齿状息肉、增生性息肉和腺瘤性息肉患者的小 RNA 转录组，以确定无蒂锯齿状息肉特有的一组独特的 microRNA (miRNA)。小 RNA 的分析将有助于区分息肉类型，类似于之前发表的使用 miRNA panel 进行的肿瘤分类研究。我们已经在每个队列中收集了 10-20 个结肠样本和 5 个血清样本，并将在研究的第一年期间在每个队列中再收集 10 个结肠样本和 15 个血清样本。根据之前的功效计算，我们相信每个队列中的 20 名患者足以定义跨息肉类型的 miRNA 差异表达。我们将使用留一法交叉验证来开发一个两步程序，用于定义无蒂锯齿状息肉患者和传统增生性息肉患者的小 RNA 基因特征。我们将通过 qPCR 在每个队列患者的组织和血清样本中验证无蒂锯齿状息肉患者独特表达的 10 个 miRNA。我们将使用 mRNA 3' UTR 和 miRNA 表达质粒的瞬时共转染、mRNA 3' UTR 的定点诱变以及 mRNA 产品的蛋白质印迹来鉴定和验证选定的 miRNA 靶标。我们的假设是，这种方法将识别分子标记，准确区分锯齿状息肉病综合征和散发性患者中的无蒂锯齿状息肉与其他息肉，并提供非侵入性血清检测来识别结肠癌风险增加的患者。目的是对患者患结肠癌的风险进行分层，以指导癌症筛查测试的频率并帮助设计化学预防试验。