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。

• 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
• 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

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- 在相当一部分人脑肿瘤中，HUWE1 功能发生改变。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 将考察候选人的作用 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。