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）是一种众所周知难以治疗的实体瘤，也是最常见的肾癌形式，对传统化疗的敏感性极低。近年来，抑制血小板源性生长因子（PDGF）受体和血管内皮生长因子（VEGF）受体的受体酪氨酸激酶（RTK）抑制药物已开始用于治疗肾细胞癌。 PDGF 和 VEGF 是由于 von Hippel-Lindau (VHL) 肿瘤抑制基因的失活突变而在 RCC 中高表达的两种生长因子。 PDGFR 和 VEGFR 分别存在于肿瘤相关周细胞和内皮细胞上，但不在肿瘤细胞本身上，抑制这些受体主要导致疾病稳定。在其他实体瘤中，RTK 抑制剂治疗靶向肿瘤细胞内在激酶，从而显着减轻肿瘤负荷。然而，尚无与肾细胞癌相关的细胞内在激酶的疾病已知。我的目标是确定 RCC 上表达的癌细胞特异性激酶作为药物开发的合理靶标。通过对肾癌细胞进行磷酸化 RTK 筛选，我鉴定出了 ROR2，这是一种以前在肾细胞癌中未知的磷酸化孤儿受体酪氨酸激酶。我们实验室的初步工作表明，ROR2 不仅在 RCC 细胞中表达，而且在来自透明细胞组织学肿瘤的 RCC 细胞中以 VHL 依赖性方式表达。我想通过结合 RCC 细胞系、shRNA 敲低细胞系和缺氧免疫印迹和 qRT-PCR 技术来评估 ROR2 表达作为 VHL 丢失和缺氧诱导因子调节的目标，从而确定 ROR2 的调节状态。我想使用免疫沉淀和自磷酸化测定进一步表征 ROR2 在 RCC 中的功能作用。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。最终目标是找到可用于 RCC 治疗的潜在抑制剂，这些抑制剂利用或利用了该激酶的存在。这项研究对公众很重要，因为它代表了 RCC 的潜在重要分子靶点。本研究的长期目标不仅是分析 ROR2 在 RCC 中的重要性，而且利用所获得的 ROR2 功能知识，希望找到可用于临床治疗 RCC 的抑制剂。