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）： 该VA功绩审查更新建议的主要目标是更好地了解膀胱癌的分子发病机理，并利用这些信息来改善该疾病的诊断，预防和治疗方法。膀胱癌是美国人中第五大流行的癌症，也是每例病例治疗的最昂贵的癌症。然而，尽管膀胱癌在研究努力中却落后于许多其他癌症类型，因此尽管临床意义，临床重要性和高医疗费用，但膀胱癌一直落后于许多其他癌症类型。结果，高度可靠的诊断和预后生物标志物以及有效的治疗方式仍然很少。该优点审查奖支持的研究集中在能够推动膀胱癌形成和沿不同途径发展的分子和遗传事件上。这些方法通过以膀胱特异性和时间控制的方式表达或废除了感兴趣的基因或废除感兴趣的基因，集中在基因工程小鼠的发展和分析上。这些研究对于膀胱癌形成中的肿瘤能力或候选基因缺乏候选基因，基因 - 校园协会，膀胱中肿瘤发展的序列，不同致癌事件的协作关系，肿瘤上皮的独特背景在肿瘤中的独特背景和肿瘤中的独特背景，肿瘤的合作关系以及肿瘤中的独特背景和肿瘤的机制。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测。这些研究也有助于确定可以进一步探索临床膀胱癌治疗干预措施的潜在靶标。该更新提案建立在过去的筹资期间建立的广泛基础上，以解决几个新的，假设驱动的且高度集中的问题。具体而言，续签建议将在三个特定的目的中研究丙酮酸激酶同工型2（PKM2）的作用，在初步研究中发现，在小鼠膀胱癌模型，人膀胱癌细胞系和标本中，在膀胱肿瘤的开始和进展中高度表达。特定的目标1旨在识别有丝分裂途径中的信号，这些信号能够上调和激活PKM2的生长促进功能，PKM2是一个极为重要但探索的区域。培养的人膀胱癌细胞系将采用基于特定的基于靶标和高吞吐量的siRNA敲低方法。具体的目标2将使用互补的过表达以及敲低方法研究PKM2在膀胱肿瘤起始中的体内作用。新型的转基因小鼠系将被设计为过表达PKM2，然后越过现有的转基因线，伴有肿瘤损伤，从而提供了确定PKM2是否促进膀胱肿瘤形成的机会。此外，将使用同工型shRNA击倒PKM2，其对肿瘤生长的影响将在异种移植和原位模型中在体外和体内进行检查。特定的目标3将评估单独或与化学治疗剂结合使用天然产物的PKM2是否有助于低度无创和高级浸润性膀胱肿瘤变体的肿瘤生长缓慢。总之，这三个一系列的研究应大大增强我们对PKM2过表达在膀胱肿瘤发生和进展中的作用的理解，以及将PKM2用作诊断生物标志物和/或治疗靶靶标的膀胱癌治疗的前景。