Trafficking of the EGF receptor to the nucleus: Mechanisms and Effects

EGF 受体转运至细胞核：机制和作用

• 批准号: 8152910
• 负责人: MICHAEL H NATHANSON
• 金额: $ 7.7万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-07 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8152910
• 关键词: Binding Proteins; Brazil; Buffers; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Calcium Signaling; Calcium/calmodulin-dependent protein kinase; Cancer Patient; Caveolae; Cell Fractionation; Cell Nucleus; Cell Proliferation; Cells; Chronic; Collaborations; Colon Carcinoma; Confocal Microscopy; EGF gene; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Growth; Growth Factor; Hepatic; Hepatocellular Damage; Hepatocyte; Hepatocyte Growth Factor; Inositol; Lead; Liver; Liver Regeneration; Liver neoplasms; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Mediating; Modeling; Monitor; Movement; Natural regeneration; Nuclear; Organ; Pathway interactions; Phospholipase C; Play; Population; Predictive Value; Primary carcinoma of the liver cells; Process; Protein Isoforms; Proteins; Proto-Oncogene Protein c-met; RNA Interference; Receptor Protein-Tyrosine Kinases; Regulation; Research; Research Personnel; Resected; Role; Signal Pathway; Signal Transduction; Techniques; Testing; Time; Tissues; Tumor Biology; Tumor Cell Line; United States National Institutes of Health; Work; calmodulin-dependent protein kinase II; cancer therapy; cell growth; cell type; malignant breast neoplasm; meetings; novel; overexpression; parent grant; receptor; trafficking; tumor; tumor growth

DESCRIPTION (provided by applicant): The liver displays a unique ability to grow and regenerate. For example, complete hepatic regeneration occurs within days to weeks after two-thirds of the liver has been resected. Chronic hepatocellular damage can lead to impaired regulation of liver regeneration, which results in hepatocellular carcinoma, one of the most common malignancies in the world. The hypothesis of the parent grant is that HGF, via c-met, regulates growth in the liver by inducing InsP3-mediated Ca2+ signals within the nucleus of hepatocytes. This FIRCA application would investigate whether this is a more general mechanism of action of receptor tyrosine kinases (RTKs) across a range of tissues. Specifically, the hypothesis of this FIRCA application is that the Epidermal Growth Factor receptor (EGFR), like c-met, regulates cell growth by inducing InsP3-mediated Ca2+ signals within the nucleus, and that this action of EGFR mediates cell proliferation in common malignancies. This hypothesis will be tested through the following specific aims: 1. whether and how the EGFR reaches the nucleus in common malignancies will be determined. We will test whether a sub-population of EGFRs in caveolae traffic to the nucleus. Intracellular movement of the receptor will be monitored by as well as by cell fractionation studies. Pathways identified in liver cells will be tested in cells derived from breast, lung, prostate, and colon cancers. 2. Whether and how EGF increases Ca2+ in the nucleus will be determined. Targeted InsP3 buffers will be used to determine whether EGF, like HGF, specifically induces InsP3 formation within the nucleus. RNA interference techniques will be used to compare PLC isoforms activated by EGF and HGF, and to determine whether these PLC isoforms vary among cell types. 3. The role of nuclear Ca2+ signals in EGF-induced cell growth will be determined. We will determine whether EGF-induced cell proliferation is disrupted by blocking either (a) movement of EGFR to the nucleus, (b) EGF-induced formation Ca2+ signals in the nucleus, or (c) activation of Ca2+-dependent proteins within the nucleus, such as CaMKII. These studies will reveal how growth factors and their corresponding receptor tyrosine kinases control nuclear Ca2+ in intact cells, and identify the distinct role this may play in regulating tumor growth. This research will be performed primarily at UFMG in Brazil in collaboration with Dawidson Gomes as an extension of Project 1 of NIH P01 DK57751. PUBLIC HEALTH RELEVANCE: Growth of cells within the liver and other organs is regulated by growth factors and their receptors, known as receptor tyrosine kinases. These studies will reveal how growth factors and their corresponding receptor tyrosine kinases control signaling within the cell nucleus, and identify the distinct role this signaling pathway may play in the abnormal growth that occurs in a wide variety of tumors.

描述（申请人提供）：肝脏显示出独特的生长和再生能力。例如，完成肝脏三分之二后几天到几周内发生完整的肝脏再生。慢性肝细胞损伤可能导致肝脏再生的调节受损，这导致肝细胞癌是世界上最常见的恶性肿瘤之一。父母赠款的假设是，通过C-MET，HGF通过在肝细胞核中诱导INSP3介导的Ca2+信号来调节肝脏的生长。该FIRCA应用将研究这是否是在整个组织中受体酪氨酸激酶（RTK）的更一般的作用机理。具体而言，这种FIRCA应用的假设是表皮生长因子受体（EGFR）（例如C-MET）通过诱导核内的INSP3介导的Ca2+信号来调节细胞生长，而EGFR的这种作用介导了普通恶性肿瘤中的细胞增殖。该假设将通过以下特定目的进行检验：1。将确定EGFR是否以及如何达到常见恶性肿瘤的核。我们将测试Caveolae流量到核的EGFR的亚构造。受体的细胞内运动将通过细胞分馏研究来监测。在肝细胞中鉴定出的途径将在源自乳腺癌，肺，前列腺和结肠癌的细胞中进行测试。 2。将确定EGF是否以及如何增加核中的Ca2+。靶向INSP3缓冲区将用于确定EGF（例如HGF）是否特异性诱导了核内的INSP3形成。 RNA干扰技术将用于比较EGF和HGF激活的PLC同工型，并确定这些PLC同工型在细胞类型之间是否有所不同。 3。将确定核Ca2+信号在EGF诱导的细胞生长中的作用。我们将通过阻断EGFR向细胞核的（A）移动EGF诱导的细胞增殖，（b）核中EGF诱导的形成Ca2+信号，还是（c）激活Ca2+依赖性蛋白在细胞核内的Ca2+依赖性蛋白。这些研究将揭示生长因子及其相应的受体酪氨酸激酶如何控制完整细胞中的核Ca2+，并确定该研究在调节肿瘤生长中可能起的独特作用。这项研究将主要在巴西的UFMG与Dawidson Gomes合作，作为NIH P01 DK57751项目1的扩展。 公共卫生相关性：肝脏和其他器官内细胞的生长受到生长因子及其受体（称为受体酪氨酸激酶）的调节。这些研究将揭示生长因子及其相应的受体酪氨酸激酶控制信号在细胞核中如何确定该信号传导途径在多种肿瘤中发生的异常生长中可能起着独特的作用。