The role of alternative pre-mRNA splicing in breast cancer progression

选择性前 mRNA 剪接在乳腺癌进展中的作用

• 批准号: 8322940
• 负责人: Klemens J Hertel
• 金额: $ 3.71万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8322940
• 关键词: Alternative Splicing; Apoptosis; Bioinformatics; Breast Cancer Cell; Cancer Biology; Cancer Patient; Cancer cell line; Cell Cycle Progression; Communities; Complex; Data; Databases; Defect; Event; Expressed Sequence Tags; Gene Expression; Gene Expression Profile; Gene Targeting; Generations; Genes; Goals; Introns; Mammary Neoplasms; Mediating; Methods; Molecular; Molecular Biology; Molecular Profiling; Normal tissue morphology; Oncogenes; Outcome; Preparation; Process; RNA; RNA Splicing; Reaction; Regulation; Role; Screening procedure; Spliceosomes; Stage Grouping; Symptoms; Testing; Time; Tissue-Specific Splicing; Tissues; Tumor Tissue; Variant; Western Blotting; Work; anticancer research; cancer therapy; insight; mRNA Precursor; malignant breast neoplasm; public health relevance; research study; tumor; tumor progression; tumorigenesis

DESCRIPTION (provided by applicant): Recent work has documented that many changes in alternative pre-mRNA splicing associate with breast cancer. These observations raise the question whether changes in pre-mRNA splicing contribute to breast cancer? Another unanswered question is whether breast cancer-specific splicing events are the result of mis-regulations or whether they are the consequence of a general splicing defect in tumors? We have demonstrated that alternative splicing events associated with breast cancer are not caused by changes in the intrinsic ability of the spliceosome to remove introns. Here, we will carry out experiments to gain insights into the question whether alternative splicing contributes to breast cancer progression. In this application, we will characterize pre-mRNA splicing networks to test the hypothesis that aberrant expression of splicing regulators trigger breast cancer-specific alternative splicing. We will combine experimental analyses with bioinformatics to characterize breast cancer-specific alternative splicing. (1) Using high throughput sequencing approaches and computational analyses of EST databases we will generate and validate a comprehensive list of breast cancer-specific alternative splicing events. (2) Using real-time PCR and quantitative western blot analysis, we will test the hypothesis that splicing regulators are differentially expressed in breast cancer. We will combine these expression profiles with computational analyses of breast cancer-specific alternative splicing events and pre-mRNA splicing predictions to determine likely targets of splicing regulatory networks. The approaches taken integrate high-throughput data generation with bioinformatics and classical molecular biology methods to gain important new insights into splicing regulatory networks. The proposed experiments take advantage of the most up-to-date experimental and analytical approaches to generate a snapshot of gene expression unique to breast cancer, thus producing a wealth of data useful to the breast cancer research community. The described experiments will demonstrate whether alternative pre-mRNA splicing is a significant contributor to breast cancer biology. Introducing these molecular insights to analyses of tissues obtained from breast cancer patients of various stage groupings will indicate to what degree changes in alternative splicing contribute to breast cancer progression, thus, providing additional screening and outcome prediction possibilities. PUBLIC HEALTH RELEVANCE: Pre-mRNA splice variants have been identified for a large variety of breast cancer genes, suggesting that widespread aberrant and alternative splicing may be a consequence or even a cause of breast cancer. Here, we propose to evaluate to what degree alternative pre-mRNA splicing contributes to breast cancer progression. The value of the molecular insights gained is realized when applying such analyses to breast cancer patients of various stage groupings, as they are likely to provide additional screening and outcome prediction possibilities.

描述（由申请人提供）：最近的工作已经证明，选择性前 mRNA 剪接的许多变化与乳腺癌相关。这些观察结果提出了一个问题：mRNA 前体剪接的变化是否会导致乳腺癌？另一个悬而未决的问题是乳腺癌特异性剪接事件是否是错误调节的结果，还是肿瘤中普遍剪接缺陷的结果？我们已经证明，与乳腺癌相关的选择性剪接事件并不是由剪接体去除内含子的内在能力的变化引起的。在这里，我们将进行实验来深入了解选择性剪接是否有助于乳腺癌进展的问题。在此应用中，我们将表征前 mRNA 剪接网络，以测试剪接调节因子的异常表达触发乳腺癌特异性选择性剪接的假设。我们将结合实验分析和生物信息学来表征乳腺癌特异性的选择性剪接。 (1) 使用高通量测序方法和 EST 数据库的计算分析，我们将生成并验证乳腺癌特异性选择性剪接事件的综合列表。 (2)利用实时PCR和定量蛋白质印迹分析，我们将检验剪接调节因子在乳腺癌中差异表达的假设。我们将把这些表达谱与乳腺癌特异性选择性剪接事件和前 mRNA 剪接预测的计算分析相结合，以确定剪接调控网络的可能目标。所采取的方法将高通量数据生成与生物信息学和经典分子生物学方法相结合，以获得剪接调控网络的重要新见解。拟议的实验利用最新的实验和分析方法来生成乳腺癌特有的基因表达快照，从而产生对乳腺癌研究界有用的大量数据。所描述的实验将证明选择性前 mRNA 剪接是否对乳腺癌生物学有重要贡献。将这些分子见解引入对从不同阶段分组的乳腺癌患者获得的组织的分析中，将表明选择性剪接的变化对乳腺癌进展的贡献程度，从而提供额外的筛查和结果预测的可能性。 公共健康相关性：多种乳腺癌基因均已鉴定出前 mRNA 剪接变体，这表明广泛的异常和选择性剪接可能是乳腺癌的结果甚至原因。在这里，我们建议评估选择性前 mRNA 剪接对乳腺癌进展的影响程度。当将此类分析应用于不同阶段分组的乳腺癌患者时，所获得的分子见解的价值得以实现，因为它们可能提供额外的筛查和结果预测的可能性。