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: 8080450
• 负责人: Klemens J Hertel
• 金额: $ 15.45万
• 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/8080450
• 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）：最近的工作证明，替代前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，即MRNA剪接的变化是否导致乳腺癌？另一个未解决的问题是，乳腺癌特异性的剪接事件是否是错误调节的结果，还是它们是肿瘤中一般剪接缺陷的结果？我们已经证明，与乳腺癌相关的替代剪接事件并非是由于剪接麻体去除内含子的内在能力的变化而引起的。在这里，我们将进行实验，以了解替代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，以检验以下假设：剪接调节剂异常表达触发乳腺癌特异性剪接。我们将与生物信息学结合实验分析，以表征乳腺癌特异性替代剪接。 （1）使用EST数据库的高吞吐量测序方法和计算分析，我们将生成并验证乳腺癌特异性替代剪接事件的全面列表。 （2）使用实时PCR和定量的Western印迹分析，我们将检验以下假设：剪接调节剂在乳腺癌中差异表达。我们将将这些表达曲线与乳腺癌特异性替代剪接事件和MRNA剪接预测的计算分析相结合，以确定剪接调节网络的可能目标。采用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，以获取有关剪接调节网络的重要新见解。提出的实验利用了最新的实验和分析方法来产生对乳腺癌独有的基因表达的快照，从而产生了对乳腺癌研究界有用的大量数据。所描述的实验将证明替代性mRNA剪接是否是乳腺癌生物学的重要促进者。将这些分子见解引入对从各个阶段分组的乳腺癌患者获得的组织的分析，将表明替代剪接的程度变化有助于乳腺癌的进展，从而提供其他筛查和结果预测的可能性。 公共卫生相关性：已确定了多种乳腺癌基因的MRNA剪接变体，这表明广泛的异常和替代剪接可能是乳腺癌的结果，甚至是造成乳腺癌的原因。在这里，我们建议评估在多大程度上有助于乳腺癌进展的替代性mRNA剪接。在对各个舞台分组的乳腺癌患者应用此类分析时，获得的分子见解的价值将实现，因为它们可能会提供额外的筛查和结果预测的可能性。