Ral in human bladder cancer progression

Ral 在人类膀胱癌进展中的作用

基本信息

批准号：
7195109
负责人：
DAN THEODORESCU
金额：
$ 32.98万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-09-01 至 2008-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7195109
关键词：
1-Phosphatidylinositol 3-Kinase Actins Address Adhesions Affect Anus Back Basement membrane Biochemistry Biological Biological Assay Biology Bladder Bladder Neoplasm Bladder Urothelium Cancer cell line Cell Line Cell membrane Cells Cellular Morphology Clinical Oncology Clinical Research Complex Connective Tissue Cytoskeleton Data Development Diagnostic Disease EGF gene EGFR Protein Overexpression Endopeptidases Epidermal Growth Factor Receptor Epithelial Family Fibroblasts Foundations Freezing Funding Future GTP-Binding Proteins Genes Genetic Growth Factor Guanine Nucleotide Dissociation Inhibitors Guanosine Triphosphate Phosphohydrolases HRAS gene Human In Vitro Incidence Invaded Invasive Knockout Mice Laboratories Lead Life Malignant Neoplasms Malignant neoplasm of urinary bladder Mediating Membrane Modeling Molecular Molecular Weight Mucous Membrane Mus Muscle Mutate Mutation PTEN gene Pathway interactions Patients Peptide Hydrolases Phenotype Phosphotransferases Process Production Protein Overexpression Proteins RRAS2 gene Range Rattus Receptor Activation Receptor Signaling Regulation Research Personnel Risk Role Role playing therapy Running Second Messenger Systems Sequence Homology Signal Pathway Signal Transduction Specimen Stimulus Stress Fibers Technology Testing Time Tissue Banks Tissues Transgenic Animals Transgenic Organisms Transport Vesicles Tumor Cell Invasion Tumor Suppressor Genes United States base bladder Carcinoma cancer cell cell growth cell motility follow-up human RRAS2 protein human tissue in vivo mRNA Expression member migration novel p120 GTPase Activating Protein phospholipase C gamma polymerization prognostic programs ras GTPase-Activating Proteins receptor reconstitution response rho second messenger tool tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Background and Significance: 40% of patients presenting with "superficial" (non-muscle-invasive) bladder cancer develop the "invasive" life-threatening form of the disease during follow up. In clinical studies, overexpression of Epidermal Growth Factor Receptor (EGFR), Ha-Ras mutation and loss of tumor suppressor gene PTEN have been associated with this phenotypic tumor transition. However, the exact molecular pathway by which these genes effectively trigger or facilitate the invasive process is incompletely understood. Our original R29 hypothesized that EGFR signaling enhances bladder tumor motility in vitro and invasion in vivo and intended to determine the signaling pathways used by EGFR in this process. Since funding of the R29 in 9/97, we have made the following important observations which support the original hypothesis and address the aims of the original application: 1) EGFR and Ras inhibition diminished the motility of invasive bladder cancer cells; 2) EGF stimulates motility in non-invasive cells via PI3K and this requires activity of Rho and Ras effector Rat; 3) In non-invasive cells, baseline RalA activity is low while invasive cells have constitutively higher activation; 4) Invasive cells have low levels of RhoGDI2 expression. Reconstitution of this gene leads to diminished motility and activity of RalA but not RhoA suggesting this gene may be the first RalGDI identified to function as an invasion suppressor; 5) Inhibition of PI3K activity via PTEN reconstitution in invasive cells with inactive PTEN, results in an inhibition of orthotopic invasion in vivo and a decrease in RhoA activity. Since the overall biology of both Ral and RhoGDI2 is poorly understood, but might be critical for regulating tumor invasion in patients with bladder cancer, we propose the Guiding Hypothesis that EGF mediates bladder tumor invasion via Ral activation. We will test this hypothesis with a matrix of technologies ranging from basic biochemistry to clinical oncology to address Ral biology in human bladder cancer. These include: 1) unique paired human bladder cancer cell lines with different invasive abilities; 2) a novel organotypic bladder model allowing in vitro study of tumor invasion; 3) an orthotopic assay evaluating the effects of candidate molecules on in vivo bladder cancer invasion; 4) transgenic and knockout mice with appropriate genetic and phenotypic profiles; 5) a human tissue bank with pathologically and clinically well characterized frozen specimens. Specific Aims: 1) Determine the role and pathobiology of Ral in bladder cancer invasion in organotypic, murine orthotopie and human tumor studies; 2) Determine the regulators of Ral activation (RhoGDI2, etc..) and their effect on intracellular Ral localization and bladder cancer nfigration and invasion; 3) Determine the protein complexes associated with Ral in vitro and in vivo, including those found in human cancer. Conclusion: Completion of these specific aims will provide biologically relevant molecular information on the signaling pathways regulating bladder cancer invasion in vivo and lead to the rational development of diagnostic and prognostic tools predicting the development of invasive disease and therapies to interfere with this process in patients with superficial bladder cancer.

描述（由申请人提供）：背景和意义：40％的患者在随访期间出现“表面”（非肌肉侵入性）膀胱癌在随访期间发展了疾病的“侵入性”。在临床研究中，表皮生长因子受体（EGFR），HA-RAS突变和肿瘤抑制基因PTEN的丧失与这种表型肿瘤转变有关。但是，这些基因有效触发或促进侵入性过程的确切分子途径是不完全理解的。我们最初的R29假设EGFR信号在体外增强了膀胱肿瘤运动性，并旨在确定EGFR在此过程中使用的信号传导途径。自从9/97在R29提供资金以来，我们做出了以下重要观察结果，这些观察支持原始假设并解决了原始应用的目的：1）EGFR和RAS抑制作用减少了浸润性膀胱癌细胞的运动性； 2）EGF通过PI3K刺激非侵入性细胞的运动，这需要RHO和RAS效应大鼠的活性； 3）在非侵入性细胞中，基线RALA活性较低，而侵入性细胞具有更高的激活。 4）侵入性细胞的RHOGDI2表达水平较低。该基因的重建导致Rala的运动性和活性减少，但没有提示该基因可能是第一个被确定为发挥入侵抑制器的Ralgdi。 5）通过PTEN抑制PTEN的抑制PTEN活性细胞的无活性PTEN，导致对原位侵袭体内的抑制作用，而RhoA活性降低。由于RAL和RHOGDI2的总体生物学知识鲜为人知，但对于调节膀胱癌患者的肿瘤侵袭至关重要，我们提出了一种指导假设，即EGF通过RAL激活介导膀胱肿瘤侵袭。我们将通过从基本生物化学到临床肿瘤学到人类膀胱癌中的RAL生物学的技术来检验该假设。其中包括：1）具有不同侵入性能力的独特成对的人膀胱癌细胞； 2）一种新型的器官膀胱模型，允许体外研究肿瘤侵袭； 3）评估候选分子对体内膀胱癌入侵的影响的原始测定； 4）具有适当遗传和表型特征的转基因和敲除小鼠； 5）具有病理和临床表征良好的冷冻标本的人体组织库。具体目的：1）确定RAL在膀胱癌入侵中的作用和病理生物学在器官，鼠矫正和人类肿瘤研究中的作用； 2）确定RAL激活的调节剂（RHOGDI2等）及其对细胞内RAL定位和膀胱癌的影响； 3）确定与体外和体内RAL相关的蛋白质复合物，包括在人类癌症中发现的蛋白质复合物。结论：这些具体目标的完成将提供有关调节体内膀胱癌入侵的信号通路的生物学相关分子信息，并导致诊断和预后工具的合理发展，以预测侵入性疾病和疗法的发展，以在患者患有乳腺癌患者的这一过程中介入这一过程。