BLR&D Research Career Scientist Award Application

研究职业科学家(RCS)奖的续签申请旨在继续正在进行的研究 现任主任者在主要尿路分子发病机制方面的活动 包括膀胱癌和肾结石在内的疾病。这两种疾病在退伍军人中都非常普遍， 并造成相当大的精神和身体痛苦以及惊人的医疗费用。尽管 社会经济影响，对这些疾病的分子发病机制的研究仍然极其有限， 从而阻碍了新的诊断、治疗和预防战略的制定。在过去的几年里 在获奖期间，RCS的实验室在确定关键的基因和 膀胱癌和肾结石发生和发展的表观遗传决定因素。使用 关于膀胱癌，RCS的实验室确定了几种特定的组合遗传驱动因素 与膀胱癌的两个主要表型变异的发生和发展有关， 例如，低级别浅表乳头状肿瘤与高级别侵袭性肿瘤。使用广泛的 技术途径包括培养细胞系、基因工程小鼠、化学致癌、 对人体标本进行分析和高通量剖析，实验室发现激活的受体 酪氨酸激酶(RTK)/RAS/PI3K通路与9p21抑癌基因(如CDKN2A)的缺失 和CDKN2B)诱导低级别浅表性乳头状膀胱癌，并激活该基因 RTK/Ras/PI3K途径与Rb家族蛋白或P53途径的缺失协同作用 诱导高级别肌肉浸润性膀胱癌的效应物。在退伍军人事务部支持的另一系列研究中 RCS的实验室证明，丙酮酸激酶2(PKM2)，一个关键的酶 厌氧糖酵解或Warburg效应在大多数膀胱癌中上调，并在 膀胱癌细胞的增殖，并且PKM2的基因消融显著减少了膀胱癌的生长 体外和体内。RNAi或小分子对PKM2的抑制也大大降低了获得性抗性 膀胱癌细胞对顺铂的敏感性。这些数据和将在提案中详细说明的其他数据在 不仅提高了我们对膀胱癌发病机制的分子基础的认识，而且为我们的研究提供了新的思路。 对膀胱癌诊断和治疗的新策略的实用见解。关于肾 RCS的实验室在解开RCS的作用和机制方面继续取得重大进展 人类最丰富的尿蛋白Tamm-Horsfall蛋白(THP或尿调蛋白)在尿液中的作用 防止肾结石形成的肠道防御。THP的缺乏使动物模型更容易受到影响 到肾内钙化，通过增加的尿液过饱和度，这与早期的 人类的特发性结石形成。由于在患者中经常观察到THP缺乏症，这些数据将 对肾结石的发病机制和预防有重要意义。在过去的四年半里， RCS获得了VA优秀奖、NIH多学科计划项目(PO1)赠款和NIH RO1赠款； 发表49篇同行评审论文；带头与退伍军人事务部和非退伍军人事务部开展多个协作项目 调查人员；继续积极培训初级教员和博士后研究员；提供了重要的 为地方和国家退伍军人事务部、大学附属机构和整个研究社区提供服务。这些 取得的成就为RCS继续和扩大他对退伍军人事务部使命的贡献奠定了基础 大学附属机构帮助推进研究、教育、病人护理、指导、服务和 跨学科协作。

This renewal application for Research Career Scientist (RCS) award seeks to continue the ongoing research activities by the incumbent that have been focused on the molecular pathogenesis of major urinary tract diseases including bladder cancer and kidney stone. Both diseases are highly prevalent among the Veterans, and inflict considerable emotional and physical suffering and staggering medical expenses. Despite the socioeconomic impact, research into the molecular pathogenesis of these diseases remains extremely limited, thus hampering the development of new diagnostic, therapeutic and preventive strategies. During the last award period, the laboratory of the RCS has made significant inroads in defining the key genetic and epigenetic determinants underlying the initiation and progression of bladder cancer and kidney stone. With respect to bladder cancer, the laboratory of the RCS identified several specific combinatorial genetic drivers that are responsible for the genesis and progression of the two major phenotypic variants of bladder cancer, e.g., low-grade superficial papillary tumors versus high-grade invasive tumors. Using a broad range of technical approaches including cultured cell lines, genetically engineered mice, chemical carcinogenesis, analysis of human specimens and high-throughput profiling, the laboratory found that the activation of receptor tyrosine kinase (RTK)/RAS/PI3K pathway collaborates with the loss of 9p21 tumor suppressors (e.g., CDKN2A and CDKN2B) to induce low-grade superficial papillary bladder cancer, and that the same activated RTK/RAS/PI3K pathway collaborates with the deficiency of RB family proteins or that of the p53 pathway effectors to induce high-grade muscle-invasive bladder cancer. In another series of studies supported by a VA Merit Review award, the laboratory of the RCS demonstrated that pyruvate kinase 2 (PKM2), a key enzyme in anaerobic glycolysis or Warburg effect, is upregulated in most bladder cancers and plays a critical role in bladder cancer cell proliferation, and that genetic ablation of PKM2 markedly reduces bladder cancer growth in vitro and in vivo. Inhibition of PKM2 by RNAi or small molecules also greatly decreases the acquired resistance of bladder cancer cells to cisplatin. These and other data to be detailed in the proposal are highly significant in not only improving our understanding of the molecular bases of bladder cancer pathogenesis, but also offer practical insights into novel strategies for bladder cancer diagnostics and treatment. With respect to kidney stone, the laboratory of the RCS continued to make major progress in unraveling the role and mechanisms of action of Tamm-Horsfall protein (THP or uromodulin), the most abundant urinary protein in humans, in urinary tract defense against kidney stone formation. The deficiency of THP renders animal models more susceptible to intra-renal calcification, via increased urinary supersaturation, that strongly resembles the early stages of idiopathic stone formation in humans. Because THP deficiency is often observed in patients, such data will have major implications on kidney stone pathogenesis and prevention. In the past four and a half years, the RCS obtained a VA Merit Award, a multidisciplinary NIH Program Project (PO1) grant and an NIH RO1 grant; published 49 peer-reviewed papers; spearheaded multiple collaborative projects with VA and non-VA investigators; continued to actively train junior faculty members and postdoctoral fellows; provided important services to local and national VA, university affiliates and research community at large. These accomplishments set the stage for the RCS to continue and expand his contributions to the mission of VA and university affiliates in helping advance research, education, patient care, mentoring, services and interdisciplinary collaborations.