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）：细胞阻止增殖和进入G1捕获或G0（静止）状态的能力对于肿瘤抑制至关重要。包括P130在内的视网膜母细胞瘤（RB）家族蛋白通过未完全了解的机制中介导G0/G1在哺乳动物细胞中停滞。进入G0/G1后，将P130招募到DNA结合梦蛋白复合物（DP，RB样，E2F和MUVB核心）中。 5种蛋白（Lin9，Lin37，Lin52，Lin54和RBBP4）的MUVB核心在转录控制中起双重作用。当绑定到G0/G1中的P130时，MUVB核心形成梦中的复合物并抑制800多个细胞周期依赖性基因。当从梦中释放时，MUVB核心与BMYB结合以激活有丝分裂基因。 DYRK1A蛋白激酶通过磷酸化LIN52亚基在促进梦中的组装中起关键作用，这是P130和MUVB核心之间结合所必需的。 DYRK1A的抑制足以阻止梦式综合体的组装并覆盖G0/G1逮捕。 DYRK1A基因被映射到唐氏综合症（DS）关键区域，其上调21造成的上调导致DS患者的心理障碍。重要的是要确定DYRK1A在肿瘤抑制中的作用，因为它的潜在价值是改善DS患者的认知功能的治疗靶标。 先前的研究表明，DYRK1A在RB家族介导的G0/G1停滞以及致癌Ras引起的衰老中的重要作用。此外，生化数据表明，DYRK1A在河马肿瘤抑制途径中的LATS2激酶的下游。我们的初步分析发现，DYRK1A基因在高级卵巢癌中经常拷贝数损失，这表明DYRK1A的失活有助于卵巢癌的发病机理。我们假设由于遗传损失或异常调节（例如LATS介导的激活的损失）导致卵巢癌中的DyRK1a失活，可以通过破坏梦境复合体的组装和功能来促进卵巢癌变。我们建议使用以下策略在细胞中表征DYRK1A途径来检验这一假设。在AIM 1中，我们将确定DYRK1A的丧失是否有助于卵巢癌前体细胞的恶性转化，以及DYRK1A功能的获得是否可以抑制体外和体内卵巢癌的生长。在AIM 2中，我们将确定河马途径在调节Dyrk1a和Dream Complex中的作​​用。由于尚未完全了解细胞中DyRK1a的功能和调节，因此我们确定了DYRK1A相互作用的蛋白质。在AIM 3中，我们将表征新颖的DYRK1A相互作用蛋白质，这些蛋白已优先考虑其在发育和癌症中的潜在意义，并确定这些因素在DYRK1A的生长停滞功能中的作用。 DYRK1A调节途径的拟议的生化和功能表征将提高我们对静态的了解，提高我们对卵巢癌发病机理的理解，并确定是否可以将DYRK1A安全地靶向DS。