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与结肠癌相关的替代前mRNA加工的变化。这些观察结果表明，异常的前mRNA加工可能有助于肿瘤发生。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，并且Wnt信号传导LEF/TCF转录因子的替代mRNA同种型的产生激活细胞生长。这些观察结果强烈表明，替代前mRNA加工参与建立改变的基因表达程序，这是结肠癌的特征。在这里，我们建议，以确定在结肠癌的替代前mRNA加工的程度，并测试是否结肠癌特异性mRNA亚型提供肿瘤生长和进展的功能优势。我们将联合收割机实验分析与生物信息学相结合，以表征结肠癌特异性的替代前mRNA加工（剪接和聚腺苷酸化）。(1)使用高通量测序方法和计算分析，我们将产生一个全面的列表，结肠癌特异性的替代前mRNA加工事件。(2)使用mRNA亚型特异性敲低和过表达方法，我们将测试mRNA亚型的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的能力的假设。我们还将使用裸鼠结肠癌异种移植模型来检验候选mRNA亚型的表达改变触发肿瘤形成的假设。(3)使用基于细胞的分析和异种移植模型，我们将测试的假设，即失调的前mRNA加工因子影响蛋白质组多样性和结肠肿瘤发生。 所描述的实验将突出结肠癌特异性替代前mRNA加工事件，这可能是额外的筛选和结果预测的发展至关重要。此外，了解结肠癌特异性替代前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得结肠癌基因表达模式的快照，从而产生对结肠癌研究界有用的丰富数据。鉴定结肠癌特异性替代前mRNA加工事件与差异基因表达模式，将有助于早期检测和改善结肠癌预后的预测。最终，替代的前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.