Gene therapy for bladder hyperactivity in diabetic rats

描述（由申请人提供） 糖尿病影响全世界1亿多人。神经元 糖尿病的后果可能导致膀胱 范围从轻度到重度的功能障碍。事实上，尿失禁 发生在高达80%的糖尿病患者中，其表现包括 膀胱感觉减退、残余尿增多或逼尿肌不稳定 (i.e.,膀胱过度活动或活动过度）。这些条件具有严重的 对个人生活质量的不利影响， 以及情感上的花费。这些与糖尿病相关的膀胱功能变化 是永久性的，需要药物治疗来扭转症状。电流 医学治疗缺乏有效性和特异性。为此，我们建议 评价K通道基因治疗改善膀胱功能的疗效 与最常用的糖尿病动物模型相关的活动过度 神经病变（如参考文献所确定），即链脲佐菌素 （STZ）-糖尿病大鼠。我们将研究STZ诱导的糖尿病对 雄性和雌性大鼠的体内膀胱功能。在具体目标#1，我们将 利用排尿反射研究清醒状态下的膀胱功能， 自由活动的大鼠，从而确定那些表现出STZ诱导的动物 膀胱过度活动记录有膀胱过度活动的大鼠将接受 单次注射hSlo/pcDNA，其编码α亚基， 人maxi-K（钾）通道。在具体目标#2中，我们将在现场使用 杂交技术来建立重组体之间的关系 转基因表达（即，hSlo/pcDNA表达）与膀胱功能的关系 同样的动物。这些研究将使我们能够牢固地建立这种关系， 在同一动物体内的转染效率和器官功能之间。 在具体目标#3中，我们将利用微阵列基因芯片分析来研究 STZ-糖尿病对大鼠膀胱中基因表达的影响 在膀胱功能障碍的程度方面， vivo.此外，我们还将研究hSlo基因治疗对基因表达的影响。 表情因此，我们预计能够建立明确的 转基因表达的程度与 这些分子变化对体内膀胱功能的影响。在本提案的第二年， 我们将研究这种基因治疗的预防能力和持续时间， approach.

DESCRIPTION (provided by applicant) Diabetes mellitus affects more than 100 million people worldwide. Neuronal alterations, as a consequence of diabetes mellitus can cause bladder dysfunction ranging from mild to severe in scope. In fact, urinary incontinence occurs in up to 80 percent of diabetic patients, and the manifestations include decreased bladder sensation, increased residual urine or detrusor instability (i.e., bladder overactivity or hyperactivity). These conditions have a severely adverse effect on the quality of life of the individual, at great monetary as well as emotional expense. These diabetes-related changes in bladder function are permanent and require medical therapy to reverse the symptoms. Current medical therapies lack both efficacy and specificity. To this end, we propose to evaluate the efficacy of K channel gene therapy to ameliorate the bladder hyperactivity associated with the most commonly used animal model of diabetic neuropathy (as determined by reference citations), that is, the streptozotocin (STZ)-diabetic rat. We shall study the effects of STZ-induced diabetes on bladder function in vivo in MALE and FEMALE rats. In Specific Aim #1 we will utilize the micturition reflex to study bladder function in conscious and freely moving rats, and thereby identify those animals exhibiting STZ-induced bladder overactivity. Rats with documented bladder hyperactivity will receive a single injection of the hSlo/pcDNA, which encodes the alpha subunit of the human maxi-K (potassium) channel. In Specific Aim #2 we will utilize in situ hybridization techniques to establish the relationship between recombinant transgene expression (i.e., hSlo/pcDNA expression) and bladder function in the same animal. Such studies will permit us to firmly establish the relationship between transfection efficiency and organ function in vivo in the same animal. In Specific Aim #3, we will utilize microarray gene chip analysis to study the effects of STZ-Diabetes on gene expression in the bladder of rats that have already been characterized with respect to the degree of bladder dysfunction in vivo. Moreover, we will also examine the effects of hSlo gene therapy on gene expression. As such, we anticipate being able to establish definite relationships between the degree of transgene expression, and the effects of these molecular changes on bladder function in vivo. In year 2 of this proposal we will study the prophylactic ability and duration of this gene therapy approach.