Trans-splicing of RNA in Endometrail Stroma and Other Tissues

描述（由申请人提供）：本申请中提出的研究是基于申请人实验室的最新发现，即人子宫内膜间质肿瘤中的JAZF 1-JJAZ 1基因融合体模拟了正常子宫内膜间质细胞中这两种基因的前mRNA之间的反式剪接结果。另外的证据表明，在正常子宫内膜中，前mRNA之间的反式剪接是生理调节的，但在肿瘤中，基因融合以异常、失调的方式产生嵌合RNA。这些数据导致了这样的假设，即其他肿瘤中的基因融合可能以类似的方式促成这些肿瘤的肿瘤表型，通过在细胞和组织发育的特定间隔期间通常由反式剪接产生的嵌合RNA的持续表达。将通过五个具体目标来实现初步调查结果和上述假设。其中的前两个目标（确定反式剪接过程中JAZF 1和JJAZ 1前mRNA的核内定位以及识别对于反式剪接应该是独特的RNA Y型中间体）将通过解决反式剪接的特征来进一步证明该过程的机制。这些目标也将提供研究程序，将是有用的，在研究其他例子的反式剪接。第三个目标（分析JAZF 1和PHF 1前mRNA之间的反式剪接）将研究可能的第二种情况下，肿瘤基因融合模仿子宫内膜间质细胞的反式剪接。第四个目标（分析嵌合RNA在子宫内膜间质细胞中的功能）将阐述反式剪接在正常子宫内膜间质中的作用，第五个目标（证明反式剪接在子宫内膜间质细胞以外的细胞中的作用）将确定与某些人类肿瘤相关的几种选定的基因融合是否反映了这些肿瘤的正常组织对应物中的反式剪接。总的来说，本申请中提出的研究将拓宽正常组织中RNA反式剪接的知识，并为一类重要的DNA重排如何在人类肿瘤中发挥致癌作用提供新的见解。公共卫生相关性：JAZF 1-JJAZ 1基因融合体通常存在于人子宫内膜的某些细胞中产生的肿瘤中，并表达与正常子宫内膜基质中RNA前体之间的反式剪接产生的杂交RNA相同的杂交RNA。这项研究将调查反式剪接的机制，它在子宫内膜中的功能，子宫内膜细胞内其他基因参与这一过程，以及其他类型肿瘤中的基因融合是否反映了正常组织中RNA前体之间的反式剪接。

DESCRIPTION (provided by applicant): The research proposed in this application is based on the recent discovery in the applicant's laboratory that a JAZF1-JJAZ1 gene fusion in human endometrial stromal tumors mimics the results of trans-splicing between pre-mRNAs for these two genes in normal endometrial stromal cells. Additional evidence suggests that in the normal endometrium trans-splicing between the pre- mRNAs is physiologically regulated, but in the tumors the gene fusion produces chimeric RNA in an abnormal, dysregulated manner. These data have led to the hypothesis that fusions of genes in other tumors may contribute to the neoplastic phenotype of those tumors in a similar fashion, through the sustained expression of chimeric RNAs normally generated by trans-splicing during specific intervals in cell and tissue development. The preliminary findings and the above hypothesis will be pursued through five specific aims. The first two of these aims (determination of the intranuclear localization of JAZF1 and JJAZ1 pre-mRNAs during the trans-splicing process and identification of an RNA Y-form intermediate that should be unique for trans-splicing) will add further proof for trans-splicing by addressing features of the mechanism for this process. These aims will also provide research procedures that will be useful in studying other examples of trans-splicing. The third aim (analysis of trans-splicing between JAZF1 and PHF1 pre-mRNAs) will investigate a likely second case of a neoplastic gene fusion mimicking trans-splicing in endometrial stromal cells. The fourth aim (analysis of chimeric RNA function in endometrial stromal cells) will address the role of trans-splicing in normal endometrial stroma, and the fifth aim (demonstration of trans-splicing in cells other those of endometrial stroma) will determine whether several, selected gene fusions associated with certain human tumors reflect trans-splicing in the normal tissue counterparts of those tumors. In aggregate, the research proposed in this application will broaden knowledge of RNA trans-splicing in normal tissues and provide new insights into how an important class of DNA rearrangements exerts oncogenic effects in human tumors. PUBLIC HEALTH RELEVANCE: The JAZF1-JJAZ1 gene fusion is commonly found in tumors arising in the certain cells of the human endometrium and expresses a hybrid RNA that is identical to one produced by trans-splicing between RNA precursors in normal endometrial stroma. The proposed research will investigate the mechanism trans-splicing, its function in the endometrium, the participation in this process of other genes within endometrial cells, and whether or not gene fusions in other kinds of tumors reflect trans-splicing between RNA precursors in normal tissues.