Bladder-specific Knockout and Gene Expression

膀胱特异性敲除和基因表达

• 批准号: 7632291
• 负责人: XUE-RU WU
• 金额: $ 26.45万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7632291
• 关键词: Ablation; Achondroplasia; Address; Animal Model; Apoptosis; Biological; Biological Models; Biology; Bladder; Bladder Neoplasm; Chondrocytes; Differentiation and Growth; Disease; Dose; Doxycycline; Dwarfism; Embryo; Epithelium; Fibroblast Growth Factor Receptors; Functional disorder; Gene Activation; Gene Expression; Generations; Genes; Genetic Recombination; Germ-Line Mutation; Goals; Growth; Healthcare; Human; Interstitial Cystitis; Knock-out; LacZ Genes; Lead; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Mediating; Molecular; Mus; Mutation; Neonatal; Oncogenes; Oncogenic; Papillary; Papillary Neoplasm; Pathogenesis; Pathway interactions; Pre-Clinical Model; Process; Property; Reaction Time; Reporter; Reporter Genes; Research; Response Elements; Retinoblastoma Genes; Role; Series; Signal Pathway; Signaling Molecule; Site; Skeletal Development; Somatic Mutation; System; Testing; Tetanus Helper Peptide; Tetracycline Control; Tetracyclines; Thanatophoric Dysplasia; Time; Trans-Activators; Transgenes; Transgenic Mice; Transgenic Model; Transgenic Organisms; Tumor Suppressor Genes; Tumor Suppressor Proteins; UPK2 gene; United States; Urinary tract infection; Urologic Diseases; Urothelium; Withdrawal; Work; base; cost; experience; fibroblast growth factor receptor 3; in vivo; insight; interest; knockout gene; malformation; mature animal; mouse model; mutant; novel; promoter; recombinase; skeletal; tool; tumor; tumorigenesis; tumorigenic

DESCRIPTION (provided by applicant): Bladder epithelium or urothelium is involved in major urinary tract diseases including bladder cancer, urinary tract infection and interstitial cystitis. However, research into the molecular bases of urothelial diseases has been hampered by the lack of effective model systems. Utilizing uroplakin II (UPII) gene promoter, we have succeeded over the last several years in targeting gene expression specifically into the urothelium of transgenic mice. The existing system, however, does not allow gene expression and inactivation to occur in a temporally controlled fashion, thus precluding many biological questions from being addressed. The overall goals of the current project are to develop and validate the second-generation transgenic systems that will allow urothelium-specific and inducible gene expression as well as knockout; and to utilize these novel systems to study the in vivo roles of several critical genes in urothelial growth, differentiation and tumorigenesis. Toward these goals, we plan to carry out two series of studies. In the first, we will generate transgenic lines in which the UPII promoter drives the urothelium-specific expression of a reverse tetracycline transactivator (UPll/rtTA). We will then test the feasibility and parameters of urothelium-specific and inducible gene expression by crossing UPll/rtTA mice with TRE/LacZ reporter mice so that LacZ expression is under the control of tetracycline response elements (TRE) and only occurs upon doxycycline treatment. In addition, we will establish a urothelium-specific and inducible knockout system, by crossing the UPll/rtTA mice with TRE/Cre mice. We will then test the functionality of this system by crossing the UPll/rtTA-TRE/Cre bi-transgenic mice with Rosa26 reporter mice in which the reporter genes are transcriptionally activated upon Cre expression. This system will allow inducible knockout of any genes of interest in the urothelium. In the second series of studies, we will inducibly express an activated fibroblast growth factor receptor as well as inducibly delete the retinoblastoma gene in the urothelium and test the hypothesis that the two genetic alterations induce low-grade superficial papillary tumors and high-grade invasive tumors, respectively. We will also examine the signaling pathways that these genetic alterations exploit in transforming the urothelium. These studies will generate powerful experimental tools for studying bladder biology and diseases and offer molecular insights regarding urothelial growth and tumorigenesis.

描述（申请人提供）：膀胱上皮或尿路上皮与主要泌尿道疾病有关，包括膀胱癌、尿路感染和间质性膀胱炎。然而，由于缺乏有效的模型系统，对尿路上皮疾病的分子基础的研究受到了阻碍。利用尿斑蛋白 II (UPII) 基因启动子，我们在过去几年中成功地将基因表达特异性靶向转基因小鼠的尿路上皮。然而，现有系统不允许基因表达和失活以时间受控的方式发生，从而阻碍了许多生物学问题的解决。当前项目的总体目标是开发和验证第二代转基因系统，该系统将允许尿路上皮特异性和诱导性基因表达以及敲除；并利用这些新系统来研究几个关键基因在尿路上皮生长、分化和肿瘤发生中的体内作用。为了实现这些目标，我们计划开展两个系列的研究。首先，我们将生成转基因株系，其中 UPII 启动子驱动反向四环素反式激活因子 (UPII/rtTA) 的尿路上皮特异性表达。然后，我们将通过将 UPll/rtTA 小鼠与 TRE/LacZ 报告小鼠杂交来测试尿路上皮特异性和可诱导基因表达的可行性和参数，以使 LacZ 表达处于四环素反应元件 (TRE) 的控制下，并且仅在强力霉素治疗时发生。此外，我们将通过将 UPll/rtTA 小鼠与 TRE/Cre 小鼠杂交，建立尿路上皮特异性和诱导型敲除系统。然后，我们将通过将 UPll/rtTA-TRE/Cre 双转基因小鼠与 Rosa26 报告小鼠杂交来测试该系统的功能，其中报告基因在 Cre 表达时被转录激活。该系统将允许诱导敲除尿路上皮中任何感兴趣的基因。在第二系列研究中，我们将诱导表达活化的成纤维细胞生长因子受体，并诱导删除尿路上皮中的视网膜母细胞瘤基因，并检验这两种基因改变诱导低级别浅表乳头状肿瘤和高级别浸润性肿瘤的假设， 分别。我们还将检查这些基因改变在转化尿路上皮时所利用的信号通路。这些研究将为研究膀胱生物学和疾病提供强大的实验工具，并提供有关尿路上皮生长和肿瘤发生的分子见解。