权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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(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30万例新诊断和4.1万人死亡。最近，一类新的基因表达调控因子microRNAs(MiRNAs)的功能障碍被认为与乳腺癌的发生、发展和转移有关。MiRNAs是一种短小的非编码RNA，与细胞质中3‘非翻译区(3’UTRs)的互补序列结合，抑制基因表达。根据生物信息学分析，每个miRNA被预测控制着一个基因产物网络，因此数百个转录本可能受到单个miRNA的调控。与靶3‘UTRs的配对不需要序列之间的完美匹配。由于这些元件退化和小，它们通常很难被检测到，因此绝大多数与癌症相关的miRNA靶标仍然未知。为了克服我们知识中的这些差距，我们开发了一种无偏倚的高通量筛选方法，名为3‘LIFE ASTIA(3’UTRs中基于发光的功能元件识别)。3‘LIFE以前所未有的规模系统地映射了3’UTRs中的miRNA靶标，使我们能够研究乳腺癌发生和发展过程中由miRNAs触发的遗传网络的动力学。我们将使用3‘LIFE方法在由1,880个3’UTRs组成的试验性文库中检测miRNA靶标，并探测两个与乳腺癌相关的miRNAs：let-7c和miR-10b。我们将绘制他们的相互作用网络，并将使用他们的结合要求，以便将目标扩展到整个人类转录组。然后，我们将在体内验证这些靶点，并使用人类乳腺条件性重编程细胞(HMCRC)系统将我们的结果与匹配的正常和早期乳腺癌活检组织中的转录组和miRNA变化进行比较。我们的方法1)开创了一种检测miRNA的创新方法：3‘非编码区相互作用，2)将检测由两个与乳腺癌相关的miRNAs控制的遗传网络，3)将精确定位早期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的药物靶点。