Detection and validation of miRNA targets in breast cancer

乳腺癌中 miRNA 靶标的检测和验证

基本信息

批准号：
8926892
负责人：
Marco Mangone
金额：
$ 16万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-12 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8926892
关键词：
3&apos Untranslated Regions Algorithms Automobile Driving Behavior Beryllium Binding Biochemical Biochemistry Bioinformatics Biological Biological Assay Biological Markers Biological Models Biology Biopsy Breast Cancer Early Detection Budgets Cancer Etiology Cancer Patient Cells Cloning Cytoplasm Data Destinations Detection Development Diagnosis Diagnostic Neoplasm Staging Disease Drug Targeting Elements Female Functional disorder Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Genome Genomics Goals Health Human Human Cell Line Indium Knowledge Laboratories Libraries Link Luciferases Malignant Neoplasms Mammary gland Mango - dietary Maps Methods MicroRNAs Modeling Molecular Names Neoplasm Metastasis Nucleotides Oncogenic Pathway interactions Patients Plasmids Process Proteins RNA Binding Regulator Genes Reporter Research Research Project Grants Resolution Sampling Specificity Staging System Systems Biology Techniques Testing The Cancer Genome Atlas TimeLine Tissue Culture Techniques Transcript Tumor stage Untranslated RNA Validation Woman base cancer initiation cost effective crosslinking and immunoprecipitation sequencing design experience high throughput screening human disease in vivo innovation interest luminescence malignant breast neoplasm miRNA expression profiling mortality novel research study screening tissue culture transcription factor triple-negative invasive breast carcinoma tumor tumor progression tumorigenic vector

项目摘要

DESCRIPTION (provided by applicant): Breast cancer is the most common cancer among women, and the second leading cause of cancer mortality in females in the U.S., with 300,000 new diagnoses and 41,000 fatalities annually. Recently, the dysfunction of a new class of gene expression regulators, named microRNAs (miRNAs), was linked to breast cancer initiation, progression and metastasis. miRNAs are short non-coding RNAs that bind to complementary sequences in the 3' UnTranslated Regions (3'UTRs) in the cytoplasm and repress gene expression. Based on bioinformatic analysis each miRNA is predicted to control a network of gene products, such that hundreds of transcripts are likely to be regulated by a single miRNA. The pairing with target 3'UTRs does not require a perfect match between the sequences. Since these elements are degenerate and small, they are generally difficult to detect, thus the vast majority of cancer relevant miRNA targets are still unknown. To overcome these gaps in our knowledge, we have developed an unbiased high-throughput screening method named 3'LIFE assay (Luminescence-based Identification of Functional Elements in 3'UTRs). 3'LIFE systematically maps miRNA targets in 3'UTRs with an unprecedented scale, allowing us to study the dynamics of genetic networks triggered by miRNAs during the initiation and the progression of breast cancer. We will use the 3'LIFE assay to detect miRNA targets in a pilot library composed of 1,880 3'UTRs, and probe two breast cancer relevant miRNAs: let-7c and miR-10b. We will map their network of interaction and will use their binding requirements in order to extend the targets to the entire human transcriptome. We will then validate the targets in vivo and compare our results to the transcriptome and miRNA changes in matched normal and early stage breast cancer biopsies using the human mammary conditionally reprogrammed cell (HMCRC) system. Our approach 1) pioneers an innovative method to detect miRNA:3'UTR interactions, 2) will detect the genetic network controlled by two breast cancer-relevant miRNAs, 3) will pinpoint tumor-specific miRNA and transcriptome changes of early stage primary breast cancers, and 4) will discover biomarkers and potential drug targets for early breast cancer detection and screening.

描述（由申请人提供）：乳腺癌是女性中最常见的癌症，也是美国女性癌症死亡的第二大原因，每年有 300,000 例新诊断病例和 41,000 例死亡病例。最近，一类名为 microRNA (miRNA) 的新型基因表达调节因子的功能障碍与乳腺癌的发生、进展和转移有关。 miRNA 是短的非编码 RNA，可与细胞质中 3' 非翻译区 (3'UTR) 的互补序列结合并抑制基因表达。根据生物信息学分析，每个 miRNA 预计将控制基因产物网络，因此数百个转录本可能由单个 miRNA 调节。与目标 3'UTR 的配对不需要序列之间的完美匹配。由于这些元件简并且小，通常难以检测，因此绝大多数癌症相关的 miRNA 靶标仍然未知。为了克服我们知识上的这些差距，我们开发了一种公正的高通量筛选方法，称为 3'LIFE 测定（基于发光的 3'UTR 功能元件识别）。 3'LIFE 以前所未有的规模系统地绘制了 3'UTR 中的 miRNA 靶标，使我们能够研究在乳腺癌发生和进展过程中由 miRNA 触发的遗传网络的动态。我们将使用 3'LIFE 检测来检测由 1,880 个 3'UTR 组成的试验文库中的 miRNA 靶标，并探测两种乳腺癌相关 miRNA：let-7c 和 miR-10b。我们将绘制它们的相互作用网络，并利用它们的结合要求将目标扩展到整个人类转录组。然后，我们将在体内验证目标，并将我们的结果与使用人类乳腺条件重编程细胞 (HMCRC) 系统匹配的正常和早期乳腺癌活检中的转录组和 miRNA 变化进行比较。我们的方法 1) 开创了一种检测 miRNA:3'UTR 相互作用的创新方法，2) 将检测由两种乳腺癌相关 miRNA 控制的遗传网络，3) 将查明早期原发性乳腺癌的肿瘤特异性 miRNA 和转录组变化，4）将发现用于早期乳腺癌检测和筛查的生物标志物和潜在药物靶点。