Coordinated regulation of alternative pre-mRNA processing in colon cancer

描述（由申请人提供）：最近的工作已经证明了替代pre-mRNA加工的变化与结肠癌有关。这些观察结果表明，异常的前mrna加工可能有助于肿瘤的发生。我们已经证明，几种RNA加工因子的表达水平在结肠癌中被错误调节，wnt信号LEF/TCF转录因子的替代mRNA异构体的产生激活了细胞生长。这些观察结果强烈表明，替代mrna前加工参与了结肠癌特征基因表达程序的改变。在这里，我们建议确定结肠癌中选择性前mRNA加工的程度，并测试结肠癌特异性mRNA亚型是否为肿瘤生长和进展提供功能优势。我们将结合实验分析和生物信息学来表征结肠癌特异性的替代pre-mRNA加工（剪接和聚腺苷酸化）。(1)利用高通量测序方法和计算分析，我们将生成结肠癌特异性替代pre-mRNA加工事件的综合列表。(2)利用mRNA亚型特异性敲低和过表达方法，我们将验证mRNA亚型错误表达改变结肠癌细胞表型和改变正常结肠维持适当细胞周期控制能力的假设。我们还将在裸鼠结肠癌异种移植模型中验证候选mRNA亚型表达改变触发肿瘤形成的假设。(3)利用基于细胞的分析和异种移植模型，我们将验证错误调节的mrna前加工因子影响蛋白质组学多样性和结肠肿瘤发生的假设。所描述的实验将突出结肠癌特异性替代pre-mRNA加工事件，这可能对开发额外的筛选和结果预测至关重要。此外，了解结肠癌特异性替代pre-mRNA加工的机制可能为人类结肠癌的新疗法铺平道路。在整个项目中，我们将依靠我们在癌症生物学，RNA生物学和生物信息学方面的专业知识来获取结肠癌基因表达模式的快照，从而为结肠癌研究界提供丰富的有用数据。结肠癌特异性替代pre-mRNA加工事件与差异基因表达模式的识别将对结肠癌的早期发现和改善预测预后有价值。最终，替代前mrna加工分析可能会发现结肠癌治疗的新靶点。

DESCRIPTION (provided by applicant): Recent work has documented that change in alternative pre-mRNA processing associate with colon cancer. These observations suggest that aberrant pre-mRNA processing may contribute to tumorigenesis. We have demonstrated that the expression levels of several RNA processing factors are misregulated in colon cancer and that generation of alternative mRNA isoforms of the Wnt-signaling LEF/TCF transcription factors activates cell growth. These observations strongly suggest that alternative pre-mRNA processing participates in establishing altered gene expression programs that are characteristic for colon cancer. Here we propose to determine the extent of alternative pre- mRNA processing in colon cancer and to test whether colon cancer-specific mRNA isoforms provide a functional advantage for tumor growth and progression. We will combine experimental analyses with bioinformatics to characterize colon cancer-specific alternative pre-mRNA processing (splicing and polyadenylation). (1) Using high throughput sequencing approaches and computational analyses we will generate a comprehensive list of colon cancer-specific alternative pre-mRNA processing events. (2) Using mRNA isoform-specific knockdown and overexpression approaches we will test the hypothesis that the misexpression of mRNA isoforms changes the colon cancer cell phenotype and alters the ability of normal colon to maintain proper cell cycle control. We will also test the hypothesis that the altered expression of candidate mRNA isoforms triggers tumor formation using colon cancer xenograft models in nude mice. (3) Using cell-based assays and xenograft models we will test the hypothesis that misregulated pre-mRNA processing factors influence proteomic diversity and colon tumorigenesis. The experiments described will highlight colon cancer-specific alternative pre-mRNA processing events that may be crucial for the development of additional screening and outcome predictions. Furthermore, understanding the mechanisms involved in colon cancer-specific alternative pre-mRNA processing may pave the way for novel therapies of human colon cancer. Throughout this project we will rely on our expertise in cancer biology, RNA biology, and bioinformatics to obtain snapshots of gene expression patterns in colon cancer, thus producing a wealth of data useful to the colon cancer research community. The identification of colon cancer-specific alternative pre-mRNA processing events coupled with differential gene expression patterns will be valuable for early detection and improved prediction of colon cancer prognosis. Ultimately, the alternative pre-mRNA processing analysis may uncover new targets for colon cancer therapy.