The Huwe1 ubiquitin ligase regulates mitosis, genomic stability and oncogenesis.

Huwe1 泛素连接酶调节有丝分裂、基因组稳定性和肿瘤发生。

基本信息

批准号：
10188467
负责人：
Antonio Iavarone
金额：
$ 37.06万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10188467
关键词：
APC gene Acceleration Acute Address Aneuploidy Biochemical Brain Brain Neoplasms Cell division Cells Chromosomal Instability Chromosome Segregation Chromosome Structures Chromosome abnormality Chromosomes Collection DNA Damage DNA Sequence Alteration Defect Development Dissection Epigenetic Process Event Gene Dosage Generations Genetic Genetic Identity Genetic Transcription Genome Genome Stability Genomic Instability Glioma Goals Human In Vitro Insertional Mutagenesis Intestines Knockout Mice Knowledge Lesion MYCN gene Malignant - descriptor Mediating Methods Mitosis Mitotic Mitotic Checkpoint Molecular Mus Mutation Neoplastic Cell Transformation Nervous system structure Neurons Normal Cell Oncogenes Oncogenic Oncoproteins Partner in relationship Patients Phenotype Play Proteins Proteome Proteomics Regulation Resistance Role Sleeping Beauty Time Transposase Tumor Suppressor Genes Tumor Suppressor Proteins Work cancer cell carcinogenesis chromosome missegregation comparative conditional knockout experimental study genome integrity in vitro Model in vivo intestinal tumorigenesis loss of function mouse model neoplastic cell neurogenesis novel preservation protein degradation segregation self-renewal stem cell differentiation tumor tumor initiation tumor progression tumorigenesis ubiquitin ligase

项目摘要

Abstract Progression from a non-transformed normal cell to a malignant cancer cell requires multiple genetic changes that activate oncogenes while restraining tumor suppressors. This occurs via the acquisition of genetic mutations and changes of gene copy number. Whole chromosome and segmental aneuploidy can alter copy number of relevant oncogenes and tumor suppressors and perturbation of the machinery controlling the fidelity of chromo- some segregation during mitosis is sufficient to promote tumor development via the generation of aneuploidy. The HECT-domain ubiquitin ligase HUWE1 coordinates the proteasomal-mediated degradation of proteins with crucial roles in cell division, differentiation and survival and recent work has shown that inactivation of HUWE1 contributes to tumor development. Initially, we generated and studied conditional knockout mice in which HUWE1 inactivation was targeted to the nervous system. We used this mouse model to characterize the role of HUWE1 in neurogenesis. We found that the activation of MYCN oncoprotein and MYCN-dependent transcriptional net- works are crucial events operating downstream of HUWE1 and whose de-regulation leads to developmental aberrations in the HUWE1-null brain. We also determined that genetic and epigenetic mechanisms drive loss- of-function alterations of HUWE1 in a significant fraction of human brain tumors. More recently, our work has revealed novel and unexpected functions of HUWE1 that have defined this ubiquitin ligase as crucial regulator of mitotic fidelity whereby loss of HUWE1 induces chromosome mis-segregation and an unstable genome. The present proposal embodies three Specific Aims that will investigate the novel functions of HUWE1 and will serve as entry point into the tumor suppressive mechanisms of HUWE1. In Aim 1 we will investigate the events that are controlled by HUWE1 to preserve mitotic fidelity and genome integrity. Aim 2 will examine the role of candi- date HUWE1 substrates that we have identified through unbiased proteomic methods for the accumulation the mitotic phenotype initiated by loss of HUWE1 activity. In Aim 3 we will proceed to address the full scope of genetic lesions that cooperate with the accumulation of DNA damage and the mitotic phenotype triggered by HUWE1 deletion to initiate brain carcinogenesis using the Sleeping Beauty transposase insertional mutagenesis.

抽象的从未转化的正常细胞发展为恶性癌细胞需要多种基因变化激活癌基因，同时抑制抑癌基因。这是通过获得基因突变而发生的以及基因拷贝数的变化。全染色体和节段非整倍体可以改变染色体的拷贝数相关癌基因和肿瘤抑制因子以及控制染色体保真度的机制的扰动有丝分裂期间的一些分离足以通过非整倍体的产生促进肿瘤的发展。 HECT 域泛素连接酶 HUWE1 协调蛋白酶体介导的蛋白质降解 HUWE1 在细胞分裂、分化和生存中发挥着至关重要的作用，最近的工作表明，HUWE1 失活有助于肿瘤的发展。最初，我们生成并研究了条件敲除小鼠，其中 HUWE1 失活是针对神经系统的。我们使用这个小鼠模型来表征 HUWE1 的作用在神经发生中。我们发现 MYCN 癌蛋白和 MYCN 依赖性转录网络的激活作品是 HUWE1 下游运行的关键事件，其放松管制会导致发展 HUWE1 缺失大脑中的畸变。我们还确定遗传和表观遗传机制会导致损失- HUWE1 在大部分人脑肿瘤中发生功能性改变。最近，我们的工作已揭示了 HUWE1 的新颖且意想不到的功能，将这种泛素连接酶定义为关键的调节因子有丝分裂保真度，HUWE1 的缺失会导致染色体错误分离和不稳定的基因组。这目前的提案体现了三个具体目标，将研究 HUWE1 的新颖功能并服务于作为 HUWE1 肿瘤抑制机制的切入点。在目标 1 中，我们将调查以下事件：由 HUWE1 控制以保持有丝分裂保真度和基因组完整性。目标 2 将检验候选人的作用日期 HUWE1 底物，我们通过无偏见的蛋白质组学方法鉴定出用于积累有丝分裂表型由 HUWE1 活性丧失引发。在目标 3 中，我们将继续解决以下问题的全部范围：与 DNA 损伤累积和有丝分裂表型相配合的遗传损伤 HUWE1 缺失通过睡美人转座酶插入突变引发脑癌。