Gene therapy for bladder hyperactivity in diabetic rats

描述(由申请人提供) 糖尿病影响着全球1亿多人。神经元 糖尿病引起的改变会导致膀胱 功能障碍范围从轻微到严重不等。事实上，尿失禁 高达80%的糖尿病患者会发生，其表现包括 膀胱感减弱、残余尿量增加或逼尿肌不稳定 (即，膀胱过度活动或多动)。这些情况有一个严重的 对个人生活质量的不利影响，以巨大的金钱 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。当前 医学治疗既缺乏有效性，也缺乏特异性。为此，我们建议 K通道基因治疗改善膀胱功能的疗效评价 与最常用的糖尿病动物模型相关的多动 神经病(由参考文献确定)，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 雄性和雌性大鼠体内的膀胱功能。在具体目标#1中，我们将利用排尿反射来研究有意识和 自由活动的大鼠，从而识别那些表现出STZ诱导的动物 膀胱过度活跃。有记录的膀胱多动的大鼠将收到 单次注射hSlo/pcDNA，它编码人 人MAXI-K(钾)通道在具体目标#2中，我们将在现场使用 利用杂交技术建立重组人的亲缘关系 转基因表达(即hSlo/pcDNA表达)与小鼠膀胱功能 同样的动物。这样的研究将使我们能够牢固地建立这种关系 同一动物体内的转染率和器官功能之间的关系。 在具体目标3中，我们将利用微阵列基因芯片分析来研究 链脲佐菌素-糖尿病对糖尿病大鼠膀胱基因表达的影响 已经就膀胱功能障碍的程度进行了表征 活着。此外，我们还将研究hSlo基因治疗对基因表达的影响。因此，我们预计能够建立明确的 转基因表达程度与转基因效果之间的关系 这些分子变化对体内膀胱功能的影响。在本计划书的第2年中 我们将研究这种基因疗法的预防能力和持续时间。 接近。

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.