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：
8701852
负责人：
Marco Mangone
金额：
$ 19.42万
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 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8701852
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 DNA Sequence Rearrangement Data Destinations Detection Development Diagnosis Diagnostic Neoplasm Staging Disease Drug Targeting Elements Female Functional RNA Functional disorder Gene Expression Gene Expression Profile Gene Targeting Genes Genetic Genome Genomics Goals Human Human Cell Line Indium Knowledge Laboratories Libraries Link Luciferases Malignant Neoplasms Mammary gland Mango - dietary Maps Methods MicroRNAs Modeling Molecular Molecular Profiling 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 Validation Woman base cancer initiation cost effective design experience high throughput screening human disease in vivo innovation interest luminescence malignant breast neoplasm mortality novel public health relevance 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率的第二大原因，每年有300,000个新诊断和41,000人死亡。最近，一种名为microRNA（miRNA）的新型基因表达调节剂的功能障碍与乳腺癌的启动，进展和转移有关。 miRNA是短的非编码RNA，与细胞质中的3'未翻译区（3'UTR）结合的互补序列结合并抑制基因表达。基于生物信息学分析，预计每个miRNA都可以控制基因产物网络，从而使数百个转录本可能受到单个miRNA调节。与目标3'UTRS配对不需要序列之间的完美匹配。由于这些元素是退化和小的，因此通常很难检测到它们，因此绝大多数癌症相关的miRNA靶标仍然未知。为了克服这些差距，我们开发了一种名为3'Life Assay（基于发光的3'UTRS功能元素的识别）的公正的高通量筛选方法。 3'Life在3'UTRS中有系统地绘制MiRNA靶标的前所未有的量表，从而使我们能够研究miRNA在启动和乳腺癌进展过程中触发的遗传网络的动力学。我们将使用3'Life分析来检测由1,880 3'UTRS组成的试验库中的miRNA靶标，并探测两个相关的miRNA：let-7c和miR-10b。我们将绘制他们的交互网络，并将使用其绑定要求将目标扩展到整个人类转录组。然后，我们将使用人类乳腺重编程细胞（HMCRC）系统进行验证体内靶标，并将我们的结果与匹配的正常和早期乳腺癌活检中的miRNA变化进行比较。我们的方法1）先驱者一种创新的方法来检测miRNA：3'UTR相互作用，2）将检测由两个与乳腺癌相关的miRNA控制的遗传网络，3）将确定肿瘤特异性miRNA和早期阶段原发性乳腺癌的转录组变化，4）将发现生物标志物和潜在的药物靶标和早期的乳腺癌检测。