DYRK1A signaling in control of cell growth, proliferation and DNA damage repair

DYRK1A 信号传导控制细胞生长、增殖和 DNA 损伤修复

• 批准号: 8963119
• 负责人: Larisa Litovchick
• 金额: $ 34.88万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8963119
• 关键词: Affect; Binding; Biochemical; Biological Assay; Cell Cycle; Cell Cycle Arrest; Cell Proliferation; Cells; Complex; DNA Binding; DNA Repair; DNA Repair Pathway; Data; Development; Disease; Down Syndrome; Drug Targeting; Family; G1 Arrest; Genes; Genetic; Genetic Models; Genetic Transcription; Growth; Health; Human; Impairment; In Vitro; Knowledge; LATS1 gene; LATS2 gene; Life; Loss of Heterozygosity; MYBL2 gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Cell; Maps; Mass Spectrum Analysis; Mediating; Mitotic; Modeling; Oncogenic; Ovarian; Ovarian Carcinoma; Ovarian Serous Adenocarcinoma; Pathogenesis; Pathway interactions; Phosphorylation; Phosphorylation Site; Phosphotransferases; Play; Process; Protein Family; Protein Kinase; Proteins; Proteomics; Psyche structure; Recruitment Activity; Regulation; Repression; Retinoblastoma; Risk; Role; Scaffolding Protein; Signal Pathway; Signal Transduction; Site-Directed Mutagenesis; Testing; Transcriptional Regulation; Tumor Suppression; Tumor Suppressor Proteins; Tumorigenicity; Up-Regulation; Woman; Work; cancer cell; carcinogenesis; cell growth; cognitive function; human FRAP1 protein; improved; in vivo; novel; ovarian neoplasm; overexpression; precursor cell; protein complex; senescence; therapeutic target

DESCRIPTION (provided by applicant): The ability of cells to arrest proliferation and to enter a G1-arrested or G0 (quiescent) state is critical for tumor suppression. Retinoblastoma (RB) family proteins, including p130, mediate G0/G1 arrest in mammalian cells by mechanisms that are not fully understood. Upon entry into G0/G1, p130 is recruited into the DNA-binding DREAM protein complex (DP, RB-like, E2F, and MuvB core). The MuvB core of 5 proteins (LIN9, LIN37, LIN52, LIN54, and RBBP4) plays a dual role in transcriptional control. When bound to p130 in G0/G1, the MuvB core forms the DREAM complex and represses more than 800 cell cycle-dependent genes. When released from DREAM, the MuvB core binds to BMYB to activate mitotic genes. DYRK1A protein kinase plays a critical role in promoting assembly of the DREAM complex by phosphorylating the LIN52 subunit, which is required for binding between the p130 and MuvB core. Inhibition of DYRK1A is sufficient to block assembly of the DREAM complex and to override G0/G1 arrest. The DYRK1A gene is mapped to a Down syndrome (DS) critical region, and its upregulation due to trisomy 21 contributes to mental impairment in people with DS. It is important to establish the role of DYRK1A in tumor suppression because of its potential value as a therapeutic target to improve the cognitive functions of people with DS. Previous studies have revealed an essential role of DYRK1A in G0/G1 arrest, mediated by RB family, as well as in oncogenic Ras-induced senescence. Furthermore, biochemical data suggest that DYRK1A is downstream of LATS2 kinase in the Hippo tumor suppressor pathway. Our preliminary analysis found that the DYRK1A gene undergoes frequent copy number loss in high-grade ovarian carcinoma, suggesting that inactivation of DYRK1A contributes to the pathogenesis of ovarian cancer. We hypothesize that inactivation of DYRK1A in ovarian cancer due to genetic losses or aberrant regulation (such as loss of LATS-mediated activation) could promote ovarian carcinogenesis by disrupting the assembly and function of the DREAM complex. We propose to test this hypothesis by characterizing DYRK1A pathways in cells using the following strategy. In Aim 1, we will determine whether loss of DYRK1A contributes to the malignant transformation of the ovarian cancer precursor cells and whether gain of DYRK1A function can suppress the growth of ovarian cancers in vitro and in vivo. In Aim 2, we will determine the role of the Hippo pathway in regulating DYRK1A and the DREAM complex. Since the function and regulation of DYRK1A in cells is not fully understood, we identified DYRK1A-interacting proteins. In Aim 3, we will characterize novel DYRK1A-interacting proteins that have been prioritized by their potential significance in development and cancer, and establish the role of these factors in the growth arrest functions of DYRK1A. The proposed biochemical and functional characterization of DYRK1A-regulated pathways will advance our knowledge of quiescence, improve our understanding of the pathogenesis of ovarian cancer, and establish whether DYRK1A can be safely targeted to treat DS.

描述（由申请人提供）：细胞抑制增殖并进入G1停滞或G 0（静止）状态的能力对于肿瘤抑制至关重要。视网膜母细胞瘤（RB）家族蛋白，包括p130，介导的G 0/G1停滞在哺乳动物细胞的机制还不完全清楚。进入G 0/G1期后，p130被募集到DNA结合DREAM蛋白复合物（DP、RB样、E2 F和MuvB核心）中。MuvB核心的5种蛋白（LIN 9、LIN 37、LIN 52、LIN 54和RBBP 4）在转录控制中起双重作用。当在G 0/G1期与p130结合时，MuvB核心形成DREAM复合物并抑制800多个细胞周期依赖性基因。当从DREAM释放时，MuvB核心与BMYB结合以激活有丝分裂基因。 DYRK 1A蛋白激酶通过磷酸化LIN 52亚基在促进DREAM复合物的组装中起关键作用，LIN 52亚基是p130和MuvB核心之间结合所必需的。DYRK 1A的抑制足以阻断DREAM复合物的组装并覆盖G 0/G1阻滞。DYRK 1A基因被定位于唐氏综合征（DS）的关键区域，由于21三体导致其上调导致DS患者的精神障碍。确定DYRK 1A在肿瘤抑制中的作用很重要，因为它作为治疗靶点的潜在价值可以改善DS患者的认知功能。 先前的研究已经揭示了DYRK 1A在RB家族介导的G 0/G1期阻滞以及致癌Ras诱导的衰老中的重要作用。此外，生化数据表明，DYRK 1A是下游的LATS 2激酶在河马肿瘤抑制途径。我们的初步分析发现，DYRK 1A基因在高级别卵巢癌中经历频繁的拷贝数丢失，这表明DYRK 1A的失活有助于卵巢癌的发病机制。我们假设，卵巢癌中DYRK 1A的失活是由于遗传丢失或异常调控（如LATS介导的激活丢失），可能通过破坏DREAM复合物的组装和功能来促进卵巢癌的发生。我们建议使用以下策略通过表征细胞中的DYRK 1A通路来测试这一假设。在目的1中，我们将确定DYRK 1A的缺失是否有助于卵巢癌前体细胞的恶性转化，以及DYRK 1A功能的获得是否可以在体外和体内抑制卵巢癌的生长。在目标2中，我们将确定Hippo通路在调节DYRK 1A和DREAM复合物中的作用。由于DYRK 1A在细胞中的功能和调控尚未完全了解，我们鉴定了DYRK 1A相互作用蛋白。在目标3中，我们将表征新型DYRK 1A相互作用蛋白，这些蛋白在发育和癌症中的潜在意义已被优先考虑，并确定这些因子在DYRK 1A生长停滞功能中的作用。DYRK 1A调节通路的拟议生化和功能表征将推进我们对静止期的认识，提高我们对卵巢癌发病机制的理解，并确定DYRK 1A是否可以安全地靶向治疗DS。