Diversity Supplement R01----Mdm2 Alternative Splicing in DNA Damage and Cancer

多样性补充剂 R01----Mdm2 选择性剪接在 DNA 损伤和癌症中的作用

基本信息

批准号：
10599711
负责人：
Dawn S Chandler
金额：
$ 7.03万
依托单位：
RESEARCH INST NATIONWIDE CHILDREN'S HOSP
依托单位国家：
美国
项目类别：
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/10599711
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 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

项目摘要

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.

抽象的肿瘤抑制p53是基因组的典型监护人，其功能在大于大于所有人类癌症中有50％。尽管p53的突变和缺失是p53失活的主要贡献者，但阴性调节剂MDM2和MDM4（MDMX）的过表达也已知p53，因此导致癌症表型。我们的实验室表明，特定类型的细胞应力会引发产生 MDM2的剪接同工型。主要的MDM2替代同工型MDM2-ALT1也称为MDM2-B，通过在A中抑制MDM2和MDM4来主要激活p53途径主要的负面方式。矛盾的是，这种同工型在几种人类癌症中被上调小儿高级神经胶质瘤，星形胶质细胞瘤，横纹肌肉瘤（RMS）和脂肪肉瘤以及成人癌症和乳腺的癌症。因此，MDM2-ALT1在癌症中起相反的作用进展取决于其表达的上下文。在拟议的研究中，我们将研究 MDM2-Alt1对p53途径控制的基础，以更好地理解1）在癌症中产生MDM2-ALT1的机制和2）所得同工型的能力使用剪接切换寡核苷酸针对目标。我们假设致癌MDM2-的表达 Alt1在肿瘤发生过程中蛋白质和RNA核因子的改变调节并且可以针对诱导剪接变化。我们将使用识别和测量剪接调节的测定法结合鉴定RNA序列及其各自的核因子的基因编辑方法 - 结合伙伴调节MDM2剪接所需的结合伙伴。此外，我们将在遗传上使用新颖工程的鼠标模型以及已建立的鼠标异种移植分析和新型剪接切换寡核苷酸（SSO）调节MDM2同工型水平。我们的工作将扩大我们对 RNA处理的组合调节应响应压力和癌症，并审问 MDM2同工型调制用于p53途径的合理控制。