Spatial and Temporal Induction of Calcineurin in the Urinary Bladder

描述(申请人提供)：我们已经证明，在部分膀胱出口梗阻(PBOO)后，钙调神经磷酸酶途径被激活，并且用其已知的抑制剂环孢素A(CsA)抑制会导致更有利的膀胱表型。尽管手术模式提供了好处，但它们确实有局限性：1)存在不同程度的肥厚。2)pBOO同时激活多条通路。3)膀胱壁内存在多种细胞类型，其中通路可能被激活，也可能不被激活。4)在一个细胞群体中存在由感兴趣途径引起的改变的可能性，从而在另一个细胞群体中引起改变。尽管手术模型在总结临床pBOO中发生的事件方面有好处，但手术模型的性质使得研究单个途径和细胞之间的相互作用非常困难。为了更全面地研究钙调神经磷酸酶介导膀胱壁肥大的机制(S)，我们现在提出了一种转基因策略，允许我们在尿路上皮或平滑肌细胞群体中的多西环素敏感启动子的控制下打开钙调神经磷酸酶。这将允许对该通路在pBOO后膀胱壁肥厚中所起的作用进行更精确的机制评估。此外，我们在本方案中产生的试剂鼠将适用于钙调神经磷酸酶途径活跃的其他系统(如心脏、肺、血管和神经)的研究。 公共卫生相关性：患者膀胱壁肥大通常是由于解剖阻塞或由于神经损伤引起的异常排尿，如患者脊柱裂或脊髓损伤所致。我们已经证明，在小鼠部分出口阻塞后，钙调神经磷酸酶途径被激活。这项拨款提案寻求利用现代转基因策略开发一种更精细的概念证明，即钙调神经磷酸酶有助于调节膀胱壁肥大。这个模型为我们提供了一个更好的平台，在其中展示使用环磷酰胺A(一种已知的钙调神经磷酸酶抑制剂)作为一种保护膀胱功能的治疗方法的潜在好处。早期制定的保留功能和预防终末期膀胱的药物治疗策略将为这些患者提供更好的生活质量。

DESCRIPTION (provided by applicant): We have shown that the calcineurin pathway is activated following partial Bladder Outlet Obstruction (pBOO), and that inhibition with its known inhibitor cyclosporine A (CSA) results in a more favorable bladder phenotype. Despite the benefits that surgical models offer, they do have limitations: 1) there is a variable degree of hypertrophy. 2) Multiple pathways are simultaneously activated with pBOO. 3) Multiple cell types exist within the bladder wall in which the pathway may or not be activated. 4) There is a potential for changes induced by the pathway of interest in one cell population to induce changes in another. Despite their benefits in a recapitulation of the events that take place in clinical pBOO, the nature of surgical models makes the study of individual pathways and cell to cell interactions very difficult. In order to more fully investigate the mechanism(s) by which calcineurin mediates bladder wall hypertrophy, we now propose the development of a transgenic strategy that allows us to turn on calcineurin under the control of the doxycycline sensitive promoter within the urothelium or the smooth muscle cell populations. This will allow for a more precise mechanistic assessment of the role this pathway plays in bladder wall hypertrophy following pBOO. In addition the reagent mouse that we generate in this proposal will find applicability in the study of other systems (such as cardiac, pulmonary, vascular, and neural) where the calcineurin pathway is active. PUBLIC HEALTH RELEVANCE: Bladder wall hypertrophy often develops in patients either as a result of an anatomic blockage or as a result of abnormal voiding due to nerve injury as seen patients spina bifida or spinal cord injury. We have shown that the calcineurin pathway is activated following partial outlet obstruction in the mouse. This grant proposal seeks to develop a more refined proof of concept using modern transgenic strategies that calcineurin helps mediate bladder wall hypertrophy. This model offers us and improved platform in which to demonstrate the potential benefit of using cyclopsorine A (a known calcineurin inhibitor) as a treatment to preserve bladder function. A strategy of preserving function and preventing the end stage bladder using pharmacologic manipulation that is instituted early will offer these patients an improved quality of life.