Trans-splicing of RNA in Endometrail Stroma and Other Tissues

子宫内膜基质和其他组织中 RNA 的反式剪接

• 批准号: 8246474
• 负责人: Jeffrey Sklar
• 金额: $ 32.59万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-04 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8246474
• 关键词: Address; Cell Line; Cell Nucleus; Cells; Chromosomal Rearrangement; Data; Development; Endometrial; Endometrial Stromal Cell; Endometrial Stromal Tumors; Endometrium; Gene Fusion; Gene Rearrangement; Genes; Genetic Transcription; Goals; Health; Human; Hybrids; Knowledge; Laboratories; Methods; Normal tissue morphology; Oncogenic; Phenotype; Polycomb; Procedures; Process; Proteins; RNA; RNA Precursors; Research; Role; Site; Testing; Tissues; Trans-Splicing; base; chimeric gene; endometrial stroma; fusion gene; insight; mRNA Precursor; neoplastic; tumor

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.

描述（由申请人提供）：本申请中提出的研究基于申请人实验室最近的发现，即人子宫内膜基质肿瘤中的JAZF1-JJAZ1基因融合体模拟了正常子宫内膜基质细胞中这两个基因的前体mRNA之间的反式剪接结果。其他证据表明，在正常子宫内膜中，前体 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。这些数据导致了这样的假设：其他肿瘤中的基因融合可能通过嵌合RNA的持续表达以类似的方式促成这些肿瘤的肿瘤表型，嵌合RNA通常是在细胞和组织发育的特定时间间隔内通过反式剪接产生的。初步发现和上述假设将通过五个具体目标来实现。其中前两个目标（确定反式剪接过程中 JAZF1 和 JJAZ1 前 mRNA 的核内定位，以及鉴定反式剪接中唯一的 RNA Y 型中间体）将通过解决该过程的机制特征来进一步证明反式剪接。这些目标还将提供有助于研究反式剪接的其他例子的研究程序。第三个目标（分析 JAZF1 和 PHF1 前 mRNA 之间的反式剪接）将研究可能的第二例子宫内膜基质细胞中模拟反式剪接的肿瘤基因融合。第四个目标（分析子宫内膜基质细胞中的嵌合RNA功能）将解决反式剪接在正常子宫内膜基质中的作用，第五个目标（演示子宫内膜基质细胞以外的细胞中的反式剪接）将确定与某些人类肿瘤相关的几个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。公共健康相关性：JAZF1-JJAZ1 基因融合体常见于人类子宫内膜某些细胞产生的肿瘤中，并表达一种杂合 RNA，该杂合 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。拟议的研究将调查反式剪接的机制、其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的情况，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。