Analysis of the Novel Kinase ROR2: A New Molecular Target in Renal Cell Carcinoma

新型激酶 ROR2 的分析：肾细胞癌的新分子靶点

• 批准号: 7617834
• 负责人: Tricia M Wright
• 金额: $ 2.76万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7617834
• 关键词: Agar; Binding; Biological Assay; Brain; Cell Line; Cell Proliferation; Cells; Clear Cell; Development; Disease; Effectiveness; Endothelial Cells; Endothelial Growth Factors; Gastrointestinal Stromal Tumors; Gene Expression; Goals; Growth; Growth Factor; Heart; Histology; Human; Hypoxia; Hypoxia Inducible Factor; Immune system; Immunoblotting; Immunoprecipitation; Kidney; Knowledge; Lead; Ligands; Lung; Malignant Epithelial Cell; Malignant Neoplasms; Molecular Target; Mus; Mutate; Oncogenic; Orphan; PDGFRB gene; Pericytes; Pharmaceutical Preparations; Pharmacologic Substance; Phosphotransferases; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Play; Preclinical Drug Evaluation; Protein Tyrosine Kinase; Proteins; Proto-Oncogene Protein c-kit; RORA gene; Receptor Inhibition; Receptor Protein-Tyrosine Kinases; Regulation; Renal Cell Carcinoma; Renal carcinoma; Role; Signal Pathway; Solid Neoplasm; Specimen; Supporting Cell; Techniques; Transcript; Tumor Burden; Tumor Suppressor Genes; Tyrosine Kinase Inhibitor; VHL mutation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factors; Work; Xenograft Model; angiogenesis; base; cancer cell; cell growth; cell motility; chemotherapy; inhibitor/antagonist; kinase inhibitor; neoplastic cell; novel; overexpression; receptor; response; small hairpin RNA; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Renal cell carcinoma (RCC), a notoriously hard to treat solid tumor and the most common form of kidney cancer, has minimal sensitivity to traditional chemotherapy. Recently, receptor tyrosine kinase (RTK) inhibitory drugs which inhibit platelet-derived growth factor (PDGF) receptor and vascular endothelial growth factor (VEGF) receptor have come into play for treating RCC. PDGF and VEGF are two growth factors highly expressed in RCC because of inactivating mutations of the von Hippel-Lindau (VHL) tumor suppressor gene. PDGFR and VEGFR are found on tumor associating pericytes and endothelial cells respectively, but not on the tumor cells themselves, with inhibition of these receptors predominantly causing disease stabilization. In other solid tumors, treatment with RTK inhibitors target tumor cell intrinsic kinases, resulting in dramatic reductions in tumor burdens. However, no disease associated cell intrinsic kinase is known for renal cell carcinoma. My aim was to identify a cancer cell specific kinase expressed on RCC as a rational target for pharmaceutical development. Using a phospho-RTK screen in renal carcinoma cells, I identified ROR2, a phosphorylated orphan receptor tyrosine kinase that was previously unknown in RCC. Preliminary work from our lab has shown that not only is ROR2 expressed in RCC cells, it is expressed in a VHL dependent manner in RCC cells from clear cell histology tumors. I would like to establish the regulation status of ROR2 by evaluating ROR2 expression as a target of VHL loss and hypoxia inducible factor regulation using a combination of RCC cell lines, shRNA knockdown cell lines and hypoxia with immunoblot and qRT-PCR techniques. I would like to further characterize the functional role of ROR2 in RCC using immunoprecipitation and autophosphorylation assays. To delineate the role ROR2 plays in renal cell carcinoma tumorigenesis, I will use a combination of shRNA knockdown cell lines to determine the influence of native ROR2 and overexpressed cell lines to delineate its oncogenic potential. The final objective is to find potential inhibitors that may be used for RCC treatment which take advantage of or exploit the presence of this kinase. This study is of importance to the public as it represents a potentially important molecular target for RCC. The long term goal of this study is not only to analyze the importance of ROR2 in RCC but also to use the gained knowledge of ROR2 function in the hopes of finding inhibitors that can be used clinically for treating RCC.

描述（由申请人提供）：肾细胞癌（RCC）是一种臭名昭著的治疗实体瘤，是肾癌最常见的形式，对传统化学疗法的敏感性最低。最近，抑制血小板衍生的生长因子（PDGF）受体和血管内皮生长因子（VEGF）受体的受体酪氨酸激酶（RTK）抑制性药物已开始用于治疗RCC。 PDGF和VEGF是在RCC中高度表达的两个生长因子，因为von Hippel-Lindau（VHL）肿瘤抑制基因的突变灭活。 PDGFR和VEGFR分别在肿瘤上的周细胞和内皮细胞上发现，但在肿瘤细胞本身中没有发现，这些受体主要导致疾病稳定。在其他实体瘤中，用RTK抑制剂的治疗靶向肿瘤细胞固有激酶，导致肿瘤负担的急剧减轻。但是，没有疾病相关的细胞固有激酶以肾细胞癌而闻名。我的目的是确定在RCC上表达的癌细胞特异性激酶是药物开发的合理靶标。使用肾脏癌细胞中的磷酸-RTK筛选，我鉴定了ROR2，ROR2是一种磷酸化的孤儿受体酪氨酸激酶，以前在RCC中是未知的。我们实验室的初步工作表明，ROR2不仅在RCC细胞中表达，而且在清晰细胞组织学肿瘤的RCC细胞中以VHL的方式表达。我想通过评估ROR2表达作为VHL丢失和缺氧诱导因子调节的靶标，建立ROR2的调节状态，并使用RCC细胞系，SHRNA敲低细胞系和使用免疫印迹和QRT-PCR技术的缺氧组合。我想进一步表征ROR2在RCC中使用免疫沉淀和自磷酸化测定的功能作用。为了描述ROR2在肾细胞癌肿瘤发生中的作用，我将使用shRNA敲低细胞系的组合来确定天然ROR2和过表达细胞系的影响，以描述其致癌潜力。最终目标是找到可能用于RCC治疗的潜在抑制剂，这些抑制剂利用或利用这种激酶的存在。这项研究对公众来说是重要的，因为它代表了RCC的潜在重要分子靶标。这项研究的长期目标不仅是分析ROR2在RCC中的重要性，而且还要利用获得ROR2功能的知识，以期找到可以在临床上用于治疗RCC的抑制剂。