Sphingolipid Signaling in Wilms' Tumor Cells

肾母细胞瘤细胞中的鞘脂信号传导

基本信息

批准号：
7694612
负责人：
FERNANDO A FERRER
金额：
$ 0.1万
依托单位：
CONNECTICUT CHILDREN'S MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-09-23 至 2010-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7694612
关键词：
Adult Agonist Binding Cell Line Cell Proliferation Childhood Childhood Renal Neoplasm Children&apos s Oncology Group Clinical Collaborations Complement Coupling Data Development End Point Enzymes Family Foundations G-Protein-Coupled Receptors GTP-Binding Proteins Genes Glioblastoma Growth Growth and Development function Human In Vitro Institution Invasive Investigation Kidney Kidney Neoplasms LIM Domain Kinase 1 Lead Letters Ligands Lipids Lyase Lysophospholipids Malignant Epithelial Cell Malignant Neoplasms Measurement Measures Mediating Metastatic Neoplasm to the Lung Morbidity - disease rate Neoplasm Metastasis Nephroblastoma Numbers Outcome Pathway interactions Patients Phenotype Phosphorylation Play Process Prognostic Marker Protein Overexpression Publishing RNA Interference Receptor Signaling Regulation Relative (related person)Renal Cell Carcinoma Reporting Research Research Personnel Reverse Transcriptase Polymerase Chain Reaction Rho-associated kinase Role SPHK1 enzyme Sampling Signal Transduction Signaling Protein Specimen Sphingolipids Sphingosine Sphingosine-1-Phosphate Receptor System Techniques Technology Time Toxin Tumor Angiogenesis Tumor Cell Line Western Blotting angiogenesis carcinogenesis cell behavior cell motility clinically relevant cofilin comparative in vivo inhibitor/antagonist insight interest migration mortality mouse model neoplastic cell novel novel therapeutics programs receptor receptor coupling receptor expression repository research study response rho rho GTP-Binding Proteins sphingosine 1-phosphate sphingosine kinase tumor tumor growth tumor progression

项目摘要

DESCRIPTION (provided by applicant): Pediatric and adult renal tumors are typically, angiogenic, invasive and potentially metatastic. Morbidity and mortality associated with Wilm's tumor is usually associated with metastases. Recently, the lysophospholipid, sphingosine-1-phosphate (S1P), and its associated G-protein-coupled receptors have emerged as putative regulators of angiogenesis, tumor proliferation, migration and metastasis. S1P receptor isotypic expression may be a determinant of tumor cell migration. We propose to gain mechanistic insights into the role of S1P receptor signaling in Wilm's tumor cells. In Aim 1 the expression of S1P receptors and S1P metabolizing enzymes in Wilm's tumor cell lines and clinical specimens will be evaluated using quantitative RT-PCR, western blot analysis, and immunohistochemical techniques. In Aim 2 the mechanisms of S1P signaling will be studied by S1P receptor overexpression and RNA interference. Cell lines that overexpress key SIP-metabolizing enzyme will be used to assess cell behavior in response to changes in S1P. S1P-receptor coupling to G proteins (Gi, GS, Gq and G12/13) and subsequent activation of Rho family GTPases (Rho, Rac, Cdc42) and Rho kinases have been implicated in S1P regulation of tumor cell motility. We will study the importance of these pathways and of the effectors downstream from Rho kinase, such as LIM-kinase (LIMK) and cofilin, by studying Rho and Rac activation, inhibition of Rho kinase, and the phosphorylation status of LIMK and cofilin. In Aim 3 growth, invasion and metastasis of naive cell lines and of S1P receptor-modified Wilm's cell lines in an established nude mouse model will be studied, thereby confirming and validating our ex vivo and in vitro results. Renal cell carcinoma cell lines from adult patients will be used for comparison. The results of these studies will contribute to our understanding of the processes of Wilm's tumor migration/metastasis and may offer novel therapeutic approaches for renal tumors.

描述（由申请人提供）：小儿和成年肾脏肿瘤通常是血管生成，侵入性且潜在的。与Wilm肿瘤相关的发病率和死亡率通常与转移有关。最近，溶物磷脂，鞘氨醇1-磷酸（S1P）及其相关的G蛋白偶联受体已成为血管生成，肿瘤增殖，迁移和转移的推定调节剂。 S1P受体同种型表达可能是肿瘤细胞迁移的决定因素。我们建议获得有关S1P受体信号在Wilm肿瘤细胞中的作用的机械见解。在AIM 1中，将使用定量RT-PCR，Western blot分析和免疫组织化学技术评估S1P受体和S1P代谢酶的表达和临床标本中的S1P代谢酶的表达。在AIM 2中，S1P受体的过表达和RNA干扰将研究S1P信号的机理。过表达过表达的键sip量代谢酶的细胞系将用于评估S1P变化的细胞行为。 S1P受体偶联与G蛋白（GI，GS，GQ和G12/13）以及随后的Rho家族GTPases（Rho，RAC，CDC42）和Rho激酶的激活与S1P调节有关肿瘤细胞运动的调节。我们将通过研究Rho和RAC激活，Rho激酶的抑制以及肢体和Cofilin的磷酸化状态，研究Rho激酶下游的这些途径和效应子的重要性。在AIM 3的生长中，将研究幼稚细胞系的侵袭和转移，以及在已建立的裸鼠模型中的S1P受体修饰的Wilm的细胞系的侵袭和转移，从而确认和验证了我们的离体和体外结果。来自成年患者的肾细胞癌细胞系将用于比较。这些研究的结果将有助于我们理解Wilm肿瘤迁移/转移的过程，并可能为肾肿瘤提供新颖的治疗方法。