Molecular and Genetic Studies of Bladder Tumorigenesis

膀胱肿瘤发生的分子和遗传学研究

• 批准号: 9257250
• 负责人: XUE-RU WU
• 金额: --
• 依托单位: VA NEW YORK HARBOR HLTHCARE/SYS/BROOKLYN
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-10-01 至 2017-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9257250
• 关键词: Ablation; Adult; Affect; Age-Years; American; Area; Attention; Automobile Driving; Award; Biological; Biological Markers; Bladder; Bladder Neoplasm; C-terminal; Cancer Model; Cancer cell line; Candidate Disease Gene; Cell Proliferation; Chemicals; Clinical; Complement; Data; Development; Diagnosis; Diagnostic; Disease; Down-Regulation; Engineering; Epithelium; Etiology; Event; Expenditure; Family; Family member; Female; Foundations; Funding; General Population; Genes; Genetic study; Genetically Engineered Mouse; Goals; Growth; Health; Human; In Vitro; Incidence; Length; Lesion; Libraries; Light; MAP Kinase Gene; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of urinary bladder; Mediating; Medical; Medicinal Plants; Modality; Modeling; Molecular; Molecular Genetics; Mus; Muscle; Nuclear; Nuclear Translocation; Oncogenic; PTEN gene; Papillary; Pathogenesis; Pathway interactions; Phosphorylation Site; Population; Prevalence; Preventive; Prognostic Marker; Protein Family; Protein Isoforms; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; Pyruvate Kinase; Research; Research Personnel; Research Support; Retinoblastoma Protein; Risk; Role; STAT3 gene; Series; Serine/Threonine Phosphorylation; Signal Transduction; Small Interfering RNA; Smoker; Smoking; Specimen; Spinal cord injury patients; Surveys; TP53 gene; Testing; Therapeutic; Therapeutic Intervention; Time; Tobacco smoking; Transgenic Mice; Transgenic Organisms; Tumor-Suppressor Gene Inactivation; Tumorigenicity; Tyrosine; Tyrosine Phosphorylation; UPK2 gene; United States; Urothelial Cell; Urothelial Hyperplasia; Urothelium; Ursidae Family; Variant; Veterans; Xenograft procedure; accurate diagnosis; aging population; base; cancer cell; cancer therapy; cancer type; combinatorial; cost; design; diagnostic biomarker; effective therapy; improved; in vivo; inhibitor/antagonist; interest; knock-down; mTOR inhibition; male; men; mouse model; mutant; neglect; new therapeutic target; non-smoker; novel; outcome forecast; overexpression; public health relevance; small hairpin RNA; socioeconomics; targeted treatment; therapeutic target; tool; tumor; tumor growth; tumor initiation; tumor progression; tumorigenesis; tumorigenic; validation studies

DESCRIPTION (provided by applicant): The main objectives of this VA Merit Review renewal proposal are to gain a better understanding of the molecular pathogenesis of bladder cancer and to utilize this information to devise improved diagnostic, preventive and therapeutic approaches for this disease. Bladder cancer is the fifth most prevalent cancer among the Americans and the costliest cancer to treat on a per case basis. However, despite its prevalence, clinical importance and high medical expenses, bladder cancer has historically lagged behind many other cancer types in research endeavors and hence new discoveries. As a result, highly reliable diagnostic and prognostic biomarkers and effective treatment modalities remain scarce. Research supported by this Merit Review Award has been focused on the molecular and genetic events that are capable of driving bladder cancer formation and progression along the divergent pathways. The approaches have concentrated on the development and analyses of genetically engineered mice by expressing or abrogating genes of interest or both in a bladder-specific and time-controlled manner. These studies have been highly informative regarding the tumorigenic ability or the lack thereof of candidate genes in bladder cancer formation, the gene-pathway association, the sequence of tumor development in the bladder, the collaborative relationship of different oncogenic events, the unique context of bladder epithelium in tumorigenesis and the mechanisms of tumor suppressor gene inactivation. Results from these studies underscore the need of combining different biomarkers for more accurate diagnosis and prediction of prognosis. These studies have also been instrumental in identifying potential targets that can be further explored for therapeutic intervention for bladder cancer clinically. This renewal proposal builds on the broad foundation established during the past funding period to tackle several new, hypothesis-driven and highly focused questions. Specifically, the renewal proposal will investigate, in three Specific Aims, the role of pyruvate kinase isoform 2 (PKM2), found during the preliminary studies to be highly expressed in mouse bladder cancer models, human bladder cancer cell lines and specimens, in bladder tumor initiation and progression. Specific Aim 1 is designed to identify signals in the mitogenic pathways that are capable of up-regulating and activating the growth-promoting functions of PKM2, an extremely important but under-explored area. Both specific-target-based and high throughput siRNA knock- down approaches will be employed with cultured human bladder cancer cell lines. Specific Aim 2 will investigate the in vivo role of PKM2 in bladder tumor initiation using complementary over-expression as well as knock-down approaches. Novel transgenic mouse lines will be engineered to over-express PKM2 and then be crossed to existing transgenic lines that bear tumor-precursor lesions, thus providing opportunity to determine whether PKM2 promotes bladder tumor formation. Additionally, PKM2 will be knocked down using isoform- specific shRNA and its effects on tumor growth will be examined in vitro and in vivo in xenograft and orthotopic models. Specific Aim 3 will evaluate whether inhibition of PKM2 with naturally occurring products, alone or in combination with chemotherapeutics, helps slow tumor growth in low-grade non-invasive as well as high-grade invasive bladder tumor variants. Together, these three series of studies should greatly enhance our understanding of the role of PKM2 over-expression in bladder tumorigenesis and progression and the prospect of using PKM2 as a diagnostic biomarker and/or a therapeutic target for bladder cancer treatment.

描述(由申请人提供)： 这项VA Merit Review更新建议的主要目的是更好地了解膀胱癌的分子发病机制，并利用这些信息设计出改进的诊断、预防和治疗方法。膀胱癌是美国人中第五大最常见的癌症，也是每个病例治疗成本最高的癌症。然而，尽管膀胱癌的发病率、临床重要性和高昂的医疗费用，在研究努力和新发现方面，膀胱癌在历史上一直落后于许多其他癌症类型。因此，高度可靠的诊断和预后生物标志物和有效的治疗方式仍然很少。这项由功绩回顾奖支持的研究一直专注于能够推动膀胱癌形成和发展的分子和基因事件。这些方法主要集中在基因工程小鼠的发展和分析上，方法是以特定的膀胱和时间控制的方式表达或去除感兴趣的基因，或两者兼而有之。这些研究对于膀胱癌形成中候选基因的致癌能力或缺失、基因途径的关联、膀胱癌发生的顺序、不同致癌事件之间的协同关系、膀胱上皮在肿瘤发生中的独特背景以及肿瘤抑制基因失活的机制等方面提供了丰富的信息。这些研究的结果强调了将不同的生物标志物结合起来以更准确地诊断和预测预后的必要性。这些研究也有助于确定潜在的靶点，可以进一步探索膀胱癌的临床治疗干预措施。这项更新建议建立在过去供资期间建立的广泛基础之上，以解决几个新的、假设驱动的和高度集中的问题。具体地说，更新提案将以三个特定目标研究丙酮酸激酶亚型2(PKM2)在膀胱癌发生和发展中的作用。在初步研究中，PKM2在小鼠膀胱癌模型、人膀胱癌细胞系和标本中高表达。具体目标1旨在识别有丝分裂通路中能够上调和激活PKM2生长促进功能的信号，这是一个极其重要但未被探索的区域。基于特定靶点和高通量siRNA敲除方法将用于培养的人膀胱癌细胞系。具体目的2将利用互补过表达和基因敲除方法研究PKM2在膀胱肿瘤发生中的体内作用。新的转基因小鼠品系将被设计成过表达PKM2，然后与现有的带有肿瘤前驱病变的转基因品系杂交，从而为确定PKM2是否促进膀胱肿瘤的形成提供了机会。此外，PKM2将使用异构体特异性shRNA被击倒，其对肿瘤生长的影响将在体外和体内异种移植和原位模型中进行检测。具体目标3将评估单独或与化疗药物联合使用自然产生的产物抑制PKM2是否有助于减缓低级别非侵袭性和高级别浸润性膀胱癌变种的肿瘤生长。总之，这三个系列的研究将大大加深我们对PKM2过度表达在膀胱癌发生发展中的作用的理解，以及将PKM2作为膀胱癌诊断生物标志物和/或治疗靶点的前景。