Gene therapy for bladder hyperactivity in diabetic rats

糖尿病大鼠膀胱过度活动症的基因治疗

• 批准号: 6524607
• 负责人: George Joseph Christ
• 金额: $ 16.7万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6524607
• 关键词: diabetes mellitus; disease /disorder model; gene expression; gene therapy; in situ hybridization; laboratory rat; medical complication; microarray technology; potassium channel; streptozotocin; technology /technique development; urinary bladder disorder; urination

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.

描述（由申请人提供） 糖尿病影响着全世界超过 1 亿人。神经元 糖尿病引起的变化会导致膀胱 功能障碍的范围从轻微到严重。其实，尿失禁 高达80%的糖尿病患者会发生这种情况，其表现包括 膀胱感觉减退、残余尿增多或逼尿肌不稳定 （即膀胱过度活动或多动）。这些情况严重 对个人的生活质量产生不利影响，造成巨大的经济损失 还有情感消耗。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。当前的 药物疗法缺乏疗效和特异性。为此，我们建议 评估 K 通道基因治疗改善膀胱的功效 与最常用的糖尿病动物模型相关的多动症 神经病（根据参考引文确定），即链脲佐菌素 (STZ)-糖尿病大鼠。我们将研究 STZ 诱发的糖尿病对 雄性和雌性大鼠的体内膀胱功能。在具体目标#1中，我们将利用排尿反射来研究有意识和有意识状态下的膀胱功能。 自由活动的大鼠，从而识别那些表现出 STZ 诱导的动物 膀胱过度活动症。有膀胱过度活动记录的大鼠将接受 单次注射 hSlo/pcDNA，编码 α 亚基 人类 maxi-K（钾）通道。在具体目标 #2 中，我们将就地利用 杂交技术建立重组体之间的关系 转基因表达（即 hSlo/pcDNA 表达）和膀胱功能 同一种动物。此类研究将使我们能够牢固地建立这种关系 同一动物的转染效率和体内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 STZ-糖尿病对糖尿病大鼠膀胱基因表达的影响 已经对膀胱功能障碍的程度进行了表征 体内。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。在本提案的第二年 我们将研究这种基因疗法的预防能力和持续时间 方法。