Mdm2 Alternative Splicing in DNA Damage and Cancer

Mdm2 选择性剪接在 DNA 损伤和癌症中的作用

• 批准号: 10280391
• 负责人: Dawn S Chandler
• 金额: $ 42.62万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10280391
• 关键词: Address; Adult; Alanine Transaminase; Alternative Splicing; Antineoplastic Agents; Astrocytoma; Automobile Driving; Binding; Binding Proteins; Bioinformatics; Biological Assay; Biology; Biophysics; Breast; Cancer Biology; Cell Line; Cell Proliferation; Cells; Cellular Stress; Chemistry; Childhood Glioma; Complex; Coupled; DNA Damage; Data; Dominant-Negative Mutation; Engineering; Event; Excision; Exons; Future; Gap Junctions; Generations; Genes; Genetically Engineered Mouse; Genome; Genotoxic Stress; Glioma; Goals; High-Throughput Nucleotide Sequencing; Human; In Vitro; Knowledge; Lead; Length; Lung; Lymphoma; MDM2 gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Measures; Mechanics; Mediating; Methodology; Methods; MicroRNAs; Modality; Modeling; Molecular; Mus; Mutation; Myosarcoma; Nuclear; Nuclear Protein; Nuclear RNA; Oligonucleotides; Oncogenic; Outcome; Pathway interactions; Pharmacologic Substance; Phenotype; Play; Positioning Attribute; Primary Neoplasm; Property; Protein Isoforms; Proteins; Publishing; RNA; RNA Biochemistry; RNA Processing; RNA Sequences; RNA Splicing; RNA metabolism; RNA-Binding Proteins; Regulation; Regulatory Element; Repression; Research; Rhabdomyosarcoma; Role; Sampling; Stress; System; TP53 gene; Technology; Testing; Therapeutic; Therapeutic Intervention; Tumor Suppressor Proteins; Variant; Viral; Work; Xenograft procedure; cancer therapy; cancer type; cell growth; cell type; combinatorial; design; drug discovery; exon skipping; genotoxicity; industry partner; liposarcoma; mRNA Precursor; mouse model; novel; novel therapeutic intervention; overexpression; prevent; response; therapy design; tumor; tumor progression; tumorigenesis; tumorigenic

ABSTRACT Tumor suppressor p53 is the quintessential guardian of the genome whose function is inhibited in greater than 50% of all human cancers. Though mutation and deletion of p53 are major contributors to p53 inactivation, overexpression of the negative regulators MDM2 and MDM4 (MDMX) are also known to inactivate p53, thus leading to the cancer phenotype. Our lab has shown that specific types of cell stress initiate the generation of an alternatively spliced isoform of MDM2. The predominant MDM2 alternative isoform, MDM2-ALT1 also known as MDM2-B, functions to primarily activate the p53 pathway by inhibiting MDM2 and MDM4 in a dominant negative fashion. Paradoxically, this isoform is upregulated in several human cancers, such as pediatric high-grade gliomas, astrocytomas, rhabdomyosarcomas (RMS), and liposarcomas, as well as adult cancers such as lymphomas and those of the breast. Thus, MDM2-ALT1 plays opposing roles in cancer progression dependent upon the context of its expression. In the proposed research, we will study the underpinnings of the control of the p53 pathway by MDM2-ALT1 to better understand 1) the specific mechanism by which that MDM2-ALT1 is generated in cancer and 2) the ability of the resultant isoforms to be targeted using splice-switching oligonucleotides. We hypothesize that the expression of oncogenic MDM2- ALT1 is modulated by alterations in protein and RNA nuclear factors during the progression to tumorigenesis and can be targeted to induce splicing changes. We will use assays that identify and measure splice regulation in conjunction with gene editing approaches to identify RNA sequences and their respective nuclear factor- binding partners necessary for regulation of MDM2 splicing. Furthermore, we will use novel genetically engineered mouse models as well as established mouse xenograft assays and novel splice switching oligonucleotides (SSOs) to modulate MDM2 isoform levels. Our work will broaden our knowledge of combinatorial regulation of RNA processing in response to stress and in cancer and interrogate the utility of MDM2 isoforms modulation for rational control of the p53 pathway.

摘要 肿瘤抑制因子p53是基因组的典型监护者，其功能在大于100%的情况下被抑制。 50%的人类癌症。虽然p53的突变和缺失是p53失活的主要原因， 负调节因子MDM 2和MDM 4（MDMX）的过表达也已知对p53有影响，因此 导致癌症表型。我们的实验室已经表明，特定类型的细胞应激启动了 MDM 2的可变剪接同种型。主要的MDM 2替代同种型MDM 2-ALT 1也 被称为MDM 2-B，其功能是通过抑制MDM 2和MDM 4而主要激活p53通路。 主导消极的方式。奇怪的是，这种同工型在几种人类癌症中上调，例如， 儿童高级别胶质瘤、星形细胞瘤、横纹肌肉瘤（RMS）和脂肪肉瘤，以及成人 癌症如淋巴瘤和乳腺癌。因此，MDM 2-ALT 1在癌症中起着相反的作用。 这取决于其表达的背景。在拟议的研究中，我们将研究 MDM 2-ALT 1控制p53通路的基础，以更好地理解1）特定的 MDM 2-ALT 1在癌症中产生机制和2）所得同种型 使用剪接转换寡核苷酸靶向。我们假设致癌MDM 2- ALT 1在肿瘤发生过程中受蛋白质和RNA核因子改变的调节 并且可以靶向诱导剪接变化。我们将使用鉴定和测量剪接调节的分析方法 结合基因编辑方法来鉴定RNA序列及其各自的核因子- 调节MDM 2剪接所必需的结合伴侣。此外，我们将使用新的基因 工程小鼠模型以及已建立的小鼠异种移植物测定和新型剪接转换 使用寡核苷酸（SSO）来调节MDM 2同种型水平。我们的工作将扩大我们对 RNA加工的组合调控，以应对压力和癌症，并询问的效用， MDM 2亚型调节用于合理控制p53途径。