SON-mediated RNA splicing in glioblastoma

胶质母细胞瘤中SON介导的RNA剪接

基本信息

批准号：
10621200
负责人：
Erin Eun-Young Ahn
金额：
$ 35.03万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621200
关键词：
Affect Alternative Splicing Apical Automobile Driving Biological Biology Brain Cell Cycle Progression Cells Complex DNA Binding DNA Repair Data Data Analyses Development Diagnosis Differentiation and Growth Epidermal Growth Factor Receptor Excision Exons Future Gene Abnormality Gene Expression Genes Glioblastoma Glioma Hematologic Neoplasms Heterogeneous-Nuclear Ribonucleoproteins Impairment In Vitro Introns Knowledge Malignant Neoplasms Mediating Meditation Messenger RNA Molecular Mutation Nerve Block Nuclear Oncogenic PDGFRA gene Patients Polypyrimidine Tract-Binding Protein Positioning Attribute Process Production Proteins RNA RNA Binding RNA Splicing Radiation therapy Recurrence Regulation Research Role Sampling Site Solid Neoplasm Specific qualifier value Splice-Site Mutation Spliceosomes TP53 gene Testing Therapeutic Transcript Up-Regulation Xenograft Model cell growth chemotherapy clinically significant cofactor genome-wide genome-wide analysis in vivo inhibitor knock-down nerve stem cell neural new therapeutic target non-genetic novel strategies novel therapeutic intervention patient derived xenograft model programs recruit stem cell genes stem cells targeted treatment temozolomide tumor

项目摘要

PROJECT SUMMARY Glioblastoma multiforme (GBM) is the most common and lethal brain malignancy with a median survival of only one year after diagnosis. Our current knowledge of the underlying basis of GBM centers mostly on several recurrent mutations in specific genes. However, non-genetic factors contributing to GBM development and progression are largely unknown. Due to our poor understanding of GBM biology, treatment options are limited to chemotherapy (temozolomide, TMZ) combined with radiotherapy. Thus, new therapeutic approaches are desperately needed to treat this deadly tumor. Emerging evidence has demonstrated that aberrant RNA splicing due to splice site mutations and/or splicing factor mutations drives oncogenic gene expression in multiple types of solid tumors and hematologic malignancies. In GBM, a few RNA splicing factors, including polypyrimidine tract-binding protein 1 (PTBP1) and hnRNP A2B1 have been recently identified as driving factors in oncogenic splicing, indicating that RNA splicing is a critical, yet to be explored, mechanism that governs a broad range of oncogenic gene expression. Our extensive preliminary data demonstrated that SON, a large nuclear speckle protein possessing DNA- and RNA-binding abilities, is highly upregulated in GBM patient samples, and there is a strong correlation between SON upregulation and short patient survival. We found that SON facilitates expression of PTBP1, thereby activating the PTBP1-meditated oncogenic splicing program. In contrast, SON inhibits the expression of PTBP2, a splicing factor required for neural exon inclusion and neural differentiation. We further revealed that SON regulates the intron removal process at the constitutive splice site in the PTBP1 transcript and regulates cassette exon inclusion/skipping at the alternative splice site in the PTBP2 transcript. We also demonstrated that SON knockdown markedly inhibits GBM cell growth and neural stem cell gene expression, and SON depletion renders patient-derived glioma stem cells (GSCs) sensitive to TMZ in vitro. Based on our preliminary data, we hypothesize that SON is a master RNA splicing regulator positioned at the apex of the splicing factor hierarchy that affects both constitutive and alternative RNA splicing, consequently turning on the oncogenic splicing program and blocking neural splicing. Thus, SON could represent a promising novel therapeutic target for GBM. To test this hypothesis, we propose to dissect the molecular mechanisms of SON functions in the regulation of constitutive and alternative RNA splicing in GBM (Aim 1), and to determine the therapeutic potential of targeting SON in vivo (Aim 2). Successful completion of this proposed study will significantly advance our knowledge of abnormal gene expression in GBM and provide a fundamental rationale for future endeavors to develop SON inhibitors.

项目摘要胶质母细胞瘤多形（GBM）是最常见和致命的脑恶性肿瘤，仅中位生存期诊断后一年。我们当前对GBM基础基础的了解主要是在几个基础上特定基因中的复发突变。但是，有助于GBM发展和进展在很大程度上未知。由于我们对GBM生物学的了解不足，治疗方案有限进行化学疗法（Temozolomide，TMZ）结合放射疗法。因此，新的治疗方法是迫切需要治疗这种致命的肿瘤。新兴证据表明RNA异常由于剪接位点突变和/或剪接因子突变引起的拼接驱动肿瘤基因表达多种类型的实体瘤和血液学恶性肿瘤。在GBM中，一些RNA剪接因子，包括息肉酰胺段结合蛋白1（PTBP1）和HNRNP A2B1已被确定为驾驶致癌剪接的因素，表明RNA剪接是至关重要的，尚未探索的机制控制着广泛的致癌基因表达。我们广泛的初步数据表明，儿子，具有DNA和RNA结合能力的大核斑点蛋白在GBM中高度上调患者样本，儿子上调与病人短暂的生存之间存在很强的相关性。我们发现儿子促进了PTBP1的表达程序。相比之下，SON抑制了PTBP2的表达，PTBP2是神经外显子包含所需的剪接因子和神经分化。我们进一步揭示了儿子在 PTBP1转录本中的组成型剪接位点，并调节盒子外显子包含/跳过替代方案 PTBP2转录本中的剪接位点。我们还证明了儿子敲除明显抑制了GBM细胞生长和神经干细胞基因表达，SON耗竭使患者衍生的神经胶质瘤干细胞（GSC）体外对TMZ敏感。根据我们的初步数据，我们假设儿子是主RNA 位于剪接因子层次结构顶端的剪接调节器，该构成因子层次结构影响构成和替代性RNA剪接，因此打开了致癌剪接程序并阻止神经剪接。因此，儿子可以代表GBM的有前途的新型治疗靶标。为了检验这一假设，我们提出了剖析SON在构成和替代RNA的调节中的分子机制在GBM中剪接（AIM 1），并确定靶向儿子体内的治疗潜力（AIM 2）。这项拟议研究的成功完成将大大提高我们对异常基因的了解在GBM中的表达，并为未来发展儿子抑制剂的努力提供了基本的理由。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Overexpression of the WWE domain of RNF146 modulates poly-(ADP)-ribose dynamics at sites of DNA damage.

RNF146 WWE 结构域的过度表达可调节 DNA 损伤位点的聚 (ADP)-核糖动力学。

DOI：
10.1101/2023.12.29.573650
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Al-Rahahleh,RashaQ;Saville,KateM;Andrews,JoelF;Wu,Zhijin;Koczor,ChristopherA;Sobol,RobertW
通讯作者：
Sobol,RobertW

eNEMAL, an enhancer RNA transcribed from a distal MALAT1 enhancer, promotes NEAT1 long isoform expression.

DOI：
10.1371/journal.pone.0251515
发表时间：
2021
期刊：
PloS one
影响因子：
3.7
作者：
Stone JK;Vukadin L;Ahn EE
通讯作者：
Ahn EE

DOI：
10.1212/nxg.0000000000200062
发表时间：
2023-06
期刊：
NEUROLOGY-GENETICS
影响因子：
3.1
作者：
Langford, Jordan;Vukadin, Lana;Carey, John C.;Botto, Lorenzo D.;Velinder, Matt;Mao, Rong;Miller, Christine E.;Filloux, Francis;Ahn, Eun-Young Erin
通讯作者：
Ahn, Eun-Young Erin

Editorial: DNA repair and nucleic acid therapeutics in cancer.