Bladder transcriptome in experimental inflammation

实验性炎症中的膀胱转录组

• 批准号: 6711583
• 负责人: RICARDO SABAN
• 金额: $ 27.24万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6711583
• 关键词: Bacillus Calmette Guerin vaccine; Mycobacterium bovis; antiinflammatory agents; autoimmune disorder; biotechnology; gene targeting; genetically modified animals; immunological substance; immunomodulators; immunopathology; immunotherapy; inflammation; interstitial cystitis; laboratory mouse; microarray technology; neurogenic urinary bladder disorder; nonhuman therapy evaluation; nuclear factor kappa beta; pain; pelvis; subtraction hybridization; transcription factor; urinary bladder epithelium; urinary tract disorder chemotherapy; urinary tract pharmacology

DESCRIPTION (provided by applicant): Interstitial Cystitis (IC) is a chronic bladder syndrome characterized by urinary urgency, frequency, nocturia pain and sterile urine. Although inflammation is not a universal characteristic of biopsies from IC patients, it seems that inflammation underline all major bladder pathologies including malignancy and represents a defense reaction to injury caused by physical damage, chemical substances, microorganisms or other gents. Indeed, areas of bladder inflammation are found in bladder disease including carcinoma, during chronic implantation of catheters, and an integrative part of bladder responses to intravesical Bacillus Calmette-Guerin (BCG). Although BCG has been proposed as a promising option for treatment of IC its mechanisms of action is not completely known. One of the theories is that intravesical BCG may be effective in treating by correcting an aberrant immune imbalance in the bladder, leading to long-term symptomatic improvement. It remains to be determined the nature of this immune imbalance. As nuclear factor kappaB (NFkappaB) has been described to modulate both bladder inflammation and the effects of BCG therapy, it is fair to propose, as a central hypothesis, that intravesical BCG leads to activation of NFkappaB and translation of pro- and anti-inflammatory genes that control both the immune and inflammatory system. The long-range goals of this application are to determine the gene networks involved in bladder responses to BCG therapy. To meet these goals, we will use the mouse as a vehicle for understanding basic biological questions regarding BCG and to permit rigorous control of experimental design. Transgenic mice with reporter genes expressed specifically on endothelial cells (Tie2-LacZ) and on promoter elements responsive to NFkappaB (p105-LacZ and p65-LacZ) will be used to address our hypothesis. In addition, the appropriate use of micro array technology combined with subtractive hybridization (SSH) will identify key molecules and mechanisms involved in the transition between acute and chronic inflammation in individual bladder layers. To address our central hypothesis, Aim 1 will quantify the time course alterations in bladder morphology as a consequence of acute and chronic intravesical BCG therapy. Aim 2 will test the hypothesis that NFkB plays a central role on BCG-induced bladder inflammation and Aim 3 will test the hypothesis that acute and chronic instillation of BCG induce a differential gene regulation in bladder mucosa and detrusor muscle. For this purpose, we will use micro array technology combined with suppression subtractive hybridization (SSH) to select bladder mucosa- and detrusor specific genes. Following target validation of SSH-selected genes, a custom array will be developed. This micro array will be used to determine how the bladder transcriptome is altered by BCG therapy.

描述（由申请人提供）：间质性膀胱炎（IC）是一种慢性膀胱综合征，其特征为尿急、尿频、排尿痛和无菌尿。虽然炎症不是IC患者活检的普遍特征，但似乎炎症强调了所有主要的膀胱病理，包括恶性肿瘤，并代表了对物理损伤、化学物质、微生物或其他有害物质引起的损伤的防御反应。事实上，膀胱炎症的区域在膀胱疾病中发现，包括癌，在导管的慢性植入期间，以及膀胱对膀胱内卡介苗（BCG）的反应的整体部分。虽然BCG已被提出作为治疗IC的有希望的选择，但其作用机制尚不完全清楚。其中一个理论是膀胱内BCG可能通过纠正膀胱中异常的免疫失衡而有效治疗，从而导致长期症状改善。这种免疫失衡的性质还有待确定。由于核因子κ B（NF κ B）已被描述为调节膀胱炎症和BCG治疗的效果，因此可以合理地提出，作为一个中心假设，膀胱内BCG导致NF κ B的激活以及控制免疫和炎症系统的促炎和抗炎基因的翻译。本申请的长期目标是确定膀胱对BCG治疗的反应所涉及的基因网络。为了实现这些目标，我们将使用小鼠作为理解关于BCG的基本生物学问题的工具，并允许严格控制实验设计。将使用具有在内皮细胞（Tie 2-LacZ）和对NF κ B应答的启动子元件（p105-LacZ和p65-LacZ）上特异性表达的报告基因的转基因小鼠来解决我们的假设。此外，适当使用微阵列技术结合消减杂交（SSH）将确定参与急性和慢性炎症之间的过渡在个别膀胱层的关键分子和机制。为了解决我们的中心假设，目标1将量化膀胱形态的时间过程中的变化，作为急性和慢性膀胱内BCG治疗的结果。目的2将检验NFkB在BCG诱导的膀胱炎症中起核心作用的假设，目的3将检验BCG的急性和慢性滴注诱导膀胱粘膜和逼尿肌中的差异基因调控的假设。为此，我们将使用微阵列技术结合抑制性消减杂交（SSH）来选择膀胱粘膜和逼尿肌特异性基因。在对SSH选择的基因进行靶向验证后，将开发定制阵列。该微阵列将用于确定BCG治疗如何改变膀胱转录组。