Regulation of anti-endocrine resistance of breast cancer by a network of non-codi

非codi网络对乳腺癌抗内分泌耐药性的调节

• 批准号: 8511589
• 负责人: Peter Kabos
• 金额: $ 17.17万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8511589
• 关键词: 3' Untranslated Regions; Address; Adjuvant; Antibodies; Binding; Biochemical; Bioinformatics; Biological; Biology; Breast; Breast Cancer Cell; Cancer Biology; Cancer cell line; Clinical; Clinical Skills; Colorado; Complex; Data; Development; Diagnostic; Distant; Down-Regulation; Endocrine; Environment; Estrogen Nuclear Receptor; Estrogen Receptors; Estrogen receptor positive; Estrogens; Failure; Functional RNA; Gene Expression; Gene Expression Regulation; Genes; Goals; Grant; Hormone Receptor; Hormones; Immunoprecipitation; Individual; Knowledge; Lead; Link; MCF7 cell; Malignant Neoplasms; Measurement; Mentors; Messenger RNA; Methodology; MicroRNAs; Modeling; Modification; Molecular; Molecular Profiling; Molecular and Cellular Biology; NCOA3 gene; Neoplasm Metastasis; Normal Cell; Nuclear; Pathogenesis; Patients; Physiological; Predictive Value; Proteins; RNA; RNA Sequences; RNA-Induced Silencing Complex; Radiation; Receptor Protein-Tyrosine Kinases; Recurrence; Regulation; Regulator Genes; Reporter; Repression; Research Personnel; Resistance; Role; Sampling; Small RNA; Study models; Systems Biology; Techniques; Testing; Therapeutic; Tissue Sample; Toxic effect; Training; Transcript; Translating; Ultraviolet Rays; Universities; Vertebral column; base; career development; crosslink; falls; hormone therapy; human tissue; improved; in vitro Model; in vivo; knock-down; malignant breast neoplasm; member; next generation; novel diagnostics; novel therapeutics; programs; protein complex; research study; response; skills; treatment response; tumorigenesis

DESCRIPTION (provided by applicant): Regulation of anti-endocrine resistance of breast cancer by a network of non-coding RNAs The discovery of regulatory non-coding RNAs has added a layer of complexity to understanding how precise regulation of gene expression is achieved in normal cells and how their dysregulation may contribute to pathological states including cancer. MicroRNAs (miRNAs or miRs) are the best defined class amongst these non-coding RNAs. MiRs are thought to function by binding to their target messenger RNAs in a sequence- specific fashion and cause repression. Abnormal expression of miRs has been linked to the pathogenesis of several malignancies including breast cancer. Conventional techniques employed to study miR-mRNA interactions include profiling of expression (by either microarray or high throughput sequencing), bio-informatic prediction of miR-mRNA targets, over-expression and knock-down of miRs followed by measurement of changes in transcript and protein level, and reporter-based analysis of 3' untranslated region-binding of miRs. Although these approaches have provided valuable information about miR-mRNA interactions, they fall short in directly demonstrating specific miR-mRNA interactions in vivo. To address these shortcomings, a biochemical technique of High Throughput Sequencing following Cross-Linked Immunoprecipitation (HITS-CLIP) was recently described, which directly isolates the miR-mRNA interactome. This technique relies on the ability of ultraviolet light (UV) to cross-link miRs and mRNAs to the argonaute proteins followed by the isolation of RNA- protein complexes by immune precipitation, isolation and sequencing of the RNA. We have successfully adapted this technique to define the miR-mRNA interactome of breast cancer cells. This application for a Mentored Career Development Grant seeks to apply a systems biology approach based on HITS-CLIP to define the role of miRs in resistance to anti-endocrine therapy in breast cancer. Given their high efficacy and low toxicity, anti-endocrine therapies form the backbone of treatments for all patients with estrogen receptor positive breast cancer. However, both primary and secondary resistance remains a problem. Results from other investigators have suggested that multiple miRs (including miR-221) are involved in estrogen receptor regulation in breast cancer. In aim 1, specific changes brought about by estrogen in the miR-mRNA interactome of breast cancer will be characterized. In aim 2, we will test our hypothesis that a specific network of miRs contributes to the responsiveness and / or resistance of breast cancer to anti-endocrine therapy. Specific miR network defined from the first two aims will be validated in clinical samples in aim 3. The training and mentoring program proposed under the guidance Drs. Thorburn and Elias will provide the applicant with an in depth expertise in the biology of small RNAs and breast cancer. University of Colorado provides an outstanding academic environment that combines expertise in breast cancer biology, RNA and bioinformatics. Data from these studies is hoped to contribute to a better understanding of the role of small RNAs in hormone responsiveness and resistance of breast cancer and ultimately to novel diagnostics and therapeutics.

描述(申请人提供)：通过非编码RNAs网络调节乳腺癌的抗内分泌抗性发现调节性非编码RNAs的发现增加了一层复杂性，有助于理解正常细胞如何实现对基因表达的精确调节，以及它们的失调如何导致包括癌症在内的病理状态。在这些非编码RNA中，microRNAs(miRNAs或miRs)是定义最好的类别。MIR被认为通过以序列特异性的方式与其目标信使RNA结合而发挥作用，并导致抑制。MiRs的异常表达与包括乳腺癌在内的多种恶性肿瘤的发病机制有关。用于研究miR-mRNA相互作用的传统技术包括表达谱(通过微阵列或高通量测序)、miR-mRNA靶标的生物信息学预测、miRs的过度表达和敲除然后测量转录和蛋白质水平的变化，以及基于报告的miRs 3‘非翻译区结合的分析。虽然这些方法提供了关于miR-mRNA相互作用的有价值的信息，但它们不能直接证明体内特定的miR-mRNA相互作用。为了克服这些缺点，最近报道了一种直接分离miR-mRNA相互作用组的高通量测序技术(HITS-CLIP)。这项技术依赖于紫外光(UV)将miRs和mRNAs与ArgAerte蛋白质交联的能力，然后通过免疫沉淀、分离和RNA测序来分离RNA-蛋白质复合体。我们已经成功地将这一技术应用于确定乳腺癌细胞的miR-mRNA相互作用组。这份申请职业发展助学金的申请书寻求应用基于HITS-CLIP的系统生物学方法来定义MIR在乳腺癌抗内分泌治疗抵抗中的作用。鉴于其高效和低毒性，抗内分泌疗法构成了所有雌激素受体阳性乳腺癌患者治疗的支柱。然而，一级和二级抗性仍然是一个问题。来自其他研究人员的结果表明，多个miR(包括miR-221)参与了乳腺癌的雌激素受体调节。在目标1中，将描述雌激素在乳腺癌miR-mRNA相互作用组中所引起的特异性变化。在目标2中，我们将检验我们的假设，即特定的MIR网络有助于乳腺癌对抗内分泌治疗的反应和/或抵抗。根据前两个目标定义的特定miR网络将在Aim 3的临床样本中得到验证。在Thorburn博士和Elias博士的指导下提出的培训和指导计划将为申请者提供小RNA和乳腺癌生物学方面的深入专业知识。科罗拉多大学提供了一个出色的学术环境，结合了乳腺癌生物学、核糖核酸和生物信息学方面的专业知识。来自这些研究的数据希望有助于更好地理解小RNA在乳腺癌的激素反应和抵抗中的作用，并最终用于新的诊断和治疗。