Regulation of RNA processing by the novel spliceosomal protein, TTDN1, in development and cancer

新型剪接体蛋白 TTDN1 在发育和癌症中对 RNA 加工的调节

• 批准号: 10066928
• 负责人: Brittany A Townley
• 金额: $ 3.15万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10066928
• 关键词: Age; Alternative Splicing; Amyotrophic Lateral Sclerosis; Area; Brain; C-terminal; CRISPR/Cas technology; Cancer cell line; Cell Communication; Cell Line; Cell physiology; Cells; Complex; DNA Repair; Data; Defect; Development; Dilated Cardiomyopathy; Disease; Disease model; Ensure; Female; Genetic Diseases; Genetic Transcription; Human; Inherited; Introns; Link; Liver; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of esophagus; Malignant neoplasm of prostate; Messenger RNA; Molecular; Mus; Mutation; Nucleotide Excision Repair; Oncogenic; Organ; Pathologic; Pathway interactions; Physiological; Protein Isoforms; Proteins; RNA; RNA Processing; RNA Splicing; Regulation; Reproducibility of Results; Role; Site-Directed Mutagenesis; System; Technology; Testing; Testis; Therapeutic; Time; Tissues; Transcript; Translating; Trichothiodystrophy; body system; cancer type; developmental disease; experimental study; genome integrity; genome-wide analysis; in vitro Model; innovation; interest; lariat debranching enzyme; mRNA Precursor; male; mouse model; next generation sequencing; novel; outcome forecast; overexpression; transcriptome sequencing; tumorigenesis

Abstract Conversion of primary mRNA transcripts into multiple distinct mature mRNA’s by alternative splicing promotes cell- and tissue-specific protein diversity that is necessary for normal cellular function. Abnormal alternative splicing is implicated in many diseases, including amyotrophic lateral sclerosis, dilated cardiomyopathy, and multiple cancer types, yet the mechanisms by which dysregulation of RNA processing pathways modulate alternative splicing are not well understood. How the disruption of RNA processing pathways translates to alternative splicing consequences is a rising area of interest that has recently been catalyzed by advances in next-generation sequencing technologies and genome-wide analysis. Therefore, understanding the contribution of RNA processing proteins to genome integrity is paramount to developing therapeutic approaches. These studies will define the role of the largely uncharacterized protein TTDN1 as a novel RNA processing protein. Mutations in TTDN1 are prevalent in the majority of cases of non-photosensitive trichothiodystrophy (NP-TTD), an inherited developmental disorder. NP-TTD belongs to the class of nucleotide excision repair- defective disorders, yet NP-TTD cases are considered DNA repair proficient. However, the molecular defects underlying NP-TTD are unknown. Additionally, TTDN1 is overexpressed in certain cancers, including cervical, prostate, and esophageal cancers, and this overexpression predicts a worse prognosis. My preliminary data indicate TTDN1 promotes mRNA processing by regulating the intron lariat debranching enzyme DBR1, and suggests this interaction is crucial for proper expression of alternative transcript isoforms. Importantly, deregulated DBR1 expression, concurrent with intron lariat processing defects, has been recently linked to aberrant isoform expression and oncogenesis, but the functional contribution of TTDN1 in cancer is unknown. My preliminary data strongly suggests a physical and functional link between TTDN1 and DBR1, tying together pre-mRNA processing and alternative isoform regulation. Aim 1 will determine the influence of the TTDN1-DBR1 interaction on the molecular regulation of the intron lariat processing pathway. Aim 2 will determine isoform regulation by TTDN1 in both malignant and tissue-specific settings using TTDN1-/- cancer cell lines as well as an already established TTDN1-deficient mouse model. Upon completion of these studies, the role of TTDN1 in regulating intron lariat processing will be defined, as will the consequences for isoform expression upon pathological alteration of TTDN1 expression. The identification of novel regulatory mechanisms connecting RNA processing to transcriptional integrity has broad implications for the many genetic disorders and cancers that feature defects in alternative splicing.

抽象的 通过替代剪接将主要mRNA转录本转化为多个不同的成熟mRNA 促进正常细胞功能所必需的细胞和组织特异性蛋白质多样性。异常 替代剪接涉及许多疾病，包括肌萎缩性侧面硬化症，扩张 心肌病和多种癌症类型，但是RNA处理失调的机制 途径调节替代剪接尚不清楚。 RNA处理途径的破坏如何 转化为替代剪接后果是一个关注的兴趣区域，最近已被催化 下一代测序技术和全基因组分析的进步。因此，了解 RNA加工蛋白对基因组完整性的贡献对于开发治疗方法至关重要。 这些研究将定义在很大程度上未表征的蛋白TTDN1作为新型RNA加工的作用 蛋白质。在大多数非光质敏感性毛thiododyromthrophophy的情况下，TTDN1中的突变很普遍 （NP-TTD），一种继承的发育障碍。 NP-TTD属于核丁基惊喜维修类 有缺陷的疾病，但NP-TTD病例被认为是DNA修复熟练的。但是，分子缺陷 基础NP-TTD是未知的。此外，TTDN1在某些癌症中过表达，包括子宫颈， 前列腺和食管癌，这种过表达的预后较差。我的初步数据 表明TTDN1通过反射内含子套索脱粒酶DBR1和 表明这种相互作用对于正确表达替代转录本同工型至关重要。重要的是， 放松管制的DBR1表达，与内含子肾脏处理缺陷并发，最近已与 异常的同工型表达和肿瘤发生，但是TTDN1在癌症中的功能贡献尚不清楚。 我的初步数据强烈建议TTDN1和DBR1之间的物理和功能联系，并将 前MRNA处理和替代同工型调节。 AIM 1将确定TTDN1-DBR1的影响 内含子套索处理途径的分子调节的相互作用。 AIM 2将确定同工型 TTDN1在恶性和组织特异性环境中使用TTDN1 - / - 癌细胞系调节 已经建立了TTDN1缺陷鼠标模型。完成这些研究后，TTDN1在 调节内含子套索的处理将被定义，同工型表达对 连接RNA的新型调节机制的识别 处理转录完整性对许多遗传疾病和癌症具有广泛的影响 替代剪接中的特征缺陷。