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 通过选择性剪接将初级 mRNA 转录物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 选择性剪接与许多疾病有关，包括肌萎缩侧索硬化症、扩张症 心肌病和多种癌症类型，但 RNA 加工失调的机制 调节选择性剪接的途径尚不清楚。 RNA加工途径如何被破坏 转化为选择性剪接后果是一个日益引起人们关注的领域，最近受到以下因素的催化 下一代测序技术和全基因组分析的进展。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。 这些研究将明确很大程度上未表征的蛋白质 TTDN1 作为新型 RNA 加工的作用 蛋白质。 TTDN1 突变在大多数非光敏性毛发硫营养不良症病例中普遍存在 (NP-TTD)，一种遗传性发育障碍。 NP-TTD 属于核苷酸切除修复类别- 有缺陷的疾病，但 NP-TTD 病例被认为具有 DNA 修复能力。然而，分子缺陷 潜在的 NP-TTD 未知。此外，TTDN1 在某些癌症中过度表达，包括宫颈癌、 前列腺癌和食道癌，这种过度表达预示着更差的预后。我的初步数据 表明 TTDN1 通过调节内含子套索脱支酶 DBR1 来促进 mRNA 加工，并且 表明这种相互作用对于替代转录亚型的正确表达至关重要。重要的是， DBR1 表达失调与内含子套索加工缺陷同时发生，最近与 异常同工型表达和肿瘤发生，但 TTDN1 在癌症中的功能贡献尚不清楚。 我的初步数据强烈表明 TTDN1 和 DBR1 之间存在物理和功能联系，将它们联系在一起 前 mRNA 加工和替代亚型调节。目标 1 将确定 TTDN1-DBR1 的影响 内含子套索加工途径分子调节的相互作用。目标 2 将确定同种型 使用 TTDN1-/- 癌细胞系以及 TTDN1 在恶性和组织特异性环境中的调节 已经建立了TTDN1缺陷小鼠模型。完成这些研究后，TTDN1 在 调节内含子套索加工的过程将被定义，同种型表达的后果也将被定义。 TTDN1表达的病理改变。连接RNA的新型调控机制的鉴定 转录完整性的处理对许多遗传性疾病和癌症具有广泛的影响 选择性剪接的特征缺陷。