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: 10427138
• 负责人: XUE-RU WU
• 金额: --
• 依托单位: VA MEDICAL CENTER
• 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 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427138
• 关键词: Address; Affect; Age-Years; Aging; American; Apoptosis; Attention; Autophagocytosis; Binding; Bladder; Bladder Neoplasm; Cancer cell line; Cell Proliferation; Cells; Chemicals; Chemoresistance; Cisplatin; Clinical; Clinical Management; Cultured Cells; DNA Damage; Data; Diagnosis; Diagnostic; Disease; Distant; Embryonic Development; Emotional; Enzymes; Event; Exhibits; Expenditure; FDA approved; Female; Frequencies; Funding; Gene Mutation; General Population; Genetic; Genetic Engineering; Genetic study; Genetically Engineered Mouse; Goals; Growth; Health; Human; Hypoxia; In Vitro; Incidence; Knock-out; Knowledge; Life; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of urinary bladder; Mediating; Medical; Molecular; Molecular Genetics; Mus; Muscle; Mutation; Nature; Neoplasm Metastasis; Pain; Papillary; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Prevalence; Procedures; Process; Protein Isoforms; Pyruvate Kinase; Quality of life; RAS genes; RNA Interference; RNA Splicing; Radical Cystectomy; Recording of previous events; Recurrence; Regimen; Regulation; Research; Resistance; Risk; Role; Series; Signal Pathway; Smoker; Smoking; Specimen; Spinal cord injury; Spinal cord injury patients; Surveys; Survival Rate; Testing; Therapeutic; Time; Tobacco smoking behavior; Transgenic Mice; Transgenic Organisms; Translating; Treatment Efficacy; Treatment Failure; United States; Up-Regulation; Urothelium; Variant; Veterans; Warburg Effect; Work; aerobic glycolysis; aging population; angiogenesis; cancer cell; cancer initiation; cancer recurrence; cancer stem cell; cancer therapy; carcinogenesis; cell growth; cigarette smoke; cigarette smoking; clinical practice; cost; design; effective therapy; exome; experience; improved; in vivo; inhibitor; insight; knock-down; lifetime risk; lost work time; male; men; mouse genetics; mouse model; neoplastic cell; non-smoker; novel; novel diagnostics; novel therapeutic intervention; novel therapeutics; overexpression; prognostication; response; small molecule; smoking-related cancer; socioeconomics; transcriptome; tumor; tumor growth; tumor initiation; tumor metabolism; tumor progression; tumorigenesis; tumorigenic; whole genome

Bladder cancer is very common, costly to manage but extremely poorly studied. The incidence of bladder cancer is disproportionally higher in the Veterans than in the general population because the disease mainly afflicts the aging males with a history of cigarette smoke. The risk of developing bladder cancer is even higher in patients with spinal cord injuries – a condition very prevalent in the Veterans. There are many challenges in the clinical management of bladder cancer including the unpredictable nature of recurrence and progression of low-grade non-invasive tumors, the aggressiveness of muscle-invasive tumors and their tendency to metastasize despite radical cystectomy, the high frequency of cisplatin resistance and the lack of effective therapy for metastatic disease. Patients with this disease therefore inevitably experience tremendous amount of pain and suffering, high medical expenses and low quality of life. Despite its prevalence, clinical challenges and socioeconomic ramifications, bladder cancer has received a level of attention far below many less common and deadly diseases. Until recently, relatively little research was devoted to bladder cancer and few if any new drugs were approved by the FDA to treat advanced bladder cancer. While the whole genome, exome and transcriptome profiling of human bladder cancer of late has injected new life into the research arena, many of the new findings have not been translated into fundamentally changing the clinical practice. The primary goals of this project are therefore to acquire a deep understanding the genetic and molecular events that play principal roles in bladder cancer formation and progression and then to utilize such information to develop new diagnostic and therapeutic strategies. Toward this overall goal, a range of studies were carried out during the last funding period that focused on isoform 2 of pyruvate kinase (PKM2), a key rate-limiting enzyme controlling aerobic glycolysis or Warburg effect. Major findings include that (i) PKM2 is upregulated in a vast majority of low-grade papillary and high-grade invasive bladder cancer; (ii) RNAi inhibition of PKM2 markedly reduces cell proliferation and increases apoptosis and autophagy; (iii) shikonin binds and inhibits PKM2-mediated tumor cell growth; (iv) shikonin and cisplatin together are much more inhibitory of bladder cancer growth in vitro and in vivo; (v) acquired resistance to cisplatin is strongly associated with PKM2 upregulation; and (vi) constitutive urothelium-specific knockout of PKM2 reduces the formation of low-grade papillary bladder tumors in genetically engineered mice. These and other data demonstrate the critical importance of PKM2 in bladder tumorigenesis and set a stage for work proposed in this renewal application. Three series of studies are designed to broaden and deepen our understanding of the role and mechanisms of action of PKM2 in different pathways and stages of bladder cancer and its therapeutic potentials. The first series will define the tumorigenic stage(s) during which PKM2 is indispensable. Sophisticated mouse genetic engineering approach will be employed to ablate PKM2 in a time-controlled manner before or after the formation of low-grade papillary or high-grade invasive bladder cancer. The second series will examine whether divergent mutations of the RTK-RAS-PI3K, the most prevalently altered pathway in bladder cancer, converge to upregulate PKM2. Overexpression and knockdown of specific mutations in cultured cells will be followed by expression analyses of PKM2 and RNA splicing factors. The third series will determine the functional significance of PKM2 upregulation in cisplatin-resistant bladder cancer cells and whether inhibiting PKM2 by RNAi or shikonin re- sensitize these cells to cisplatin using in vitro and in vivo approaches. Together, these three series of studies should offer novel insights into the role(s) and underlying mechanism(s) of PKM2 overexpression during bladder cancer formation and progression and the therapeutic value of inhibiting PKM2 in reducing cisplatin resistance and in treating various forms of bladder cancer.

膀胱癌非常常见，治疗成本高昂，但研究极少。膀胱的发病率 退伍军人中癌症的发病率比普通人群高得不成比例，因为这种疾病主要是 困扰有吸烟史的老年男性。患膀胱癌的风险更高 脊髓损伤患者——这种情况在退伍军人中非常普遍。存在很多挑战 膀胱癌的临床治疗，包括复发和进展的不可预测性 低度非侵袭性肿瘤、肌肉侵袭性肿瘤的侵袭性及其倾向 尽管进行了根治性膀胱切除术，但仍发生转移，顺铂耐药发生率高且缺乏有效的治疗 转移性疾病的治疗。因此，患有这种疾病的患者不可避免地会经历巨大的痛苦。 痛苦和磨难、高昂的医疗费用和低下的生活质量。尽管其普遍存在，但临床挑战 和社会经济影响，膀胱癌受到的关注程度远远低于其他国家 常见和致命的疾病。直到最近，针对膀胱癌的研究相对较少，而且很少有研究涉及膀胱癌。 FDA 批准任何新药用于治疗晚期膀胱癌。虽然全基因组、外显子组 人类膀胱癌的转录组分析最近为研究领域注入了新的活力，许多 的新发现尚未转化为从根本上改变临床实践。初级 因此，该项目的目标是深入了解发挥作用的遗传和分子事件 膀胱癌形成和进展中的主要作用，然后利用这些信息开发新的 诊断和治疗策略。为了实现这一总体目标，期间开展了一系列研究 上一个资助期重点关注丙酮酸激酶同工型 2 (PKM2)，这是一种关键的限速酶控制 有氧糖酵解或瓦伯格效应。主要发现包括 (i) PKM2 在绝大多数细胞中表达上调 低级别乳头状癌和高级别浸润性膀胱癌； (ii) PKM2 的 RNAi 抑制显着减少细胞 增殖并增加细胞凋亡和自噬； (iii)紫草素结合并抑制PKM2介导的肿瘤细胞 生长; (iv) 紫草素和顺铂一起在体外和体内对膀胱癌的生长具有更强的抑制作用 体内； (v) 对顺铂的获得性耐药与 PKM2 上调密切相关； (vi) 本质性的 尿路上皮特异性敲除 PKM2 可减少低级别膀胱乳头状肿瘤的形成 基因工程小鼠。这些和其他数据证明了 PKM2 在膀胱中的至关重要性 肿瘤发生并为本次更新申请中提出的工作奠定基础。三个系列的研究是 旨在扩大和加深我们对 PKM2 在不同疾病中的作用和作用机制的理解 膀胱癌的途径和阶段及其治疗潜力。第一个系列将定义 PKM2 不可或缺的致瘤阶段。复杂的小鼠基因工程方法 将用于在低品位形成之前或之后以时间控制的方式消融 PKM2 乳头状或高级别浸润性膀胱癌。第二个系列将检查是否存在不同的突变 RTK-RAS-PI3K 是膀胱癌中最普遍改变的通路，其聚合上调 PKM2。 培养细胞中特定突变的过度表达和敲低将进行表达分析 PKM2 和 RNA 剪接因子。第三系列将确定PKM2的功能意义 顺铂耐药膀胱癌细胞中的上调以及是否通过 RNAi 或紫草素抑制 PKM2 使用体外和体内方法使这些细胞对顺铂敏感。这三个系列的研究一起 应该对 PKM2 过度表达的作用和潜在机制提供新的见解 膀胱癌的形成、进展及抑制PKM2减少顺铂的治疗价值 抵抗力和治疗各种形式的膀胱癌。

项目成果

Chemoprevention of urothelial cell carcinoma growth and invasion by the dual COX-LOX inhibitor licofelone in UPII-SV40T transgenic mice.

UPII-SV40T转基因小鼠中双Cox-lox抑制剂licofelone对尿路上皮细胞癌的生长和侵袭的化学预防。

• DOI: 10.1158/1940-6207.capr-14-0087
• 发表时间: 2014-07
• 期刊: Cancer prevention research (Philadelphia, Pa.)
• 作者: Madka V;Mohammed A;Li Q;Zhang Y;Patlolla JM;Biddick L;Lightfoot S;Wu XR;Steele V;Kopelovich L;Rao CV
• 通讯作者: Rao CV