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最近的发现，即人子宫内膜间质肿瘤中的JAZF1-JJAZ1基因融合模拟了正常子宫内膜间质细胞中这两个基因的前mrna之间反式剪接的结果。其他证据表明，在正常子宫内膜中，前mrna之间的反式剪接受到生理调节，但在肿瘤中，基因融合以异常、失调的方式产生嵌合RNA。这些数据导致了一种假设，即其他肿瘤中的基因融合可能以类似的方式促进这些肿瘤的肿瘤表型，通过在细胞和组织发育的特定时间间隔内由反式剪接产生的嵌合rna的持续表达。初步发现和上述假设将通过五个具体目标来追求。前两个目标（确定JAZF1和JJAZ1前mrna在反式剪接过程中的核内定位，以及鉴定一种对反式剪接来说应该是独一无二的RNA y型中间物）将通过解决这一过程的机制特征，进一步证明反式剪接的存在。这些目标也将为研究反式剪接的其他例子提供有用的研究程序。第三个目的（分析JAZF1和PHF1前mrna之间的反式剪接）将研究子宫内膜基质细胞中可能出现的第二例模拟反式剪接的肿瘤基因融合。第四个目标（分析子宫内膜间质细胞中的嵌合RNA功能）将解决反式剪接在正常子宫内膜间质中的作用，第五个目标（证明子宫内膜间质细胞中的反式剪接）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正常组织中的反式剪接。总的来说，本应用程序提出的研究将拓宽对正常组织中RNA反式剪接的认识，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：JAZF1-JJAZ1基因融合通常在人类子宫内膜某些细胞中产生的肿瘤中发现，并表达一种杂交RNA，与正常子宫内膜基质中RNA前体之间反式剪接产生的RNA相同。拟开展的研究将探讨反式剪接的机制及其在子宫内膜中的作用，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.