Human Bladder Urothelial Cell Structure and Function in Bladder Hypersensation

膀胱过敏中的人膀胱尿路上皮细胞结构和功能

• 批准号: 7856568
• 负责人: TOBY C CHAI
• 金额: $ 1.41万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-16 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7856568
• 关键词: Acetylcholine; Address; Antibodies; Apical; Area; Attenuated; Basal Cell; Basic Science; Biology; Biopsy; Bladder; Bladder Urothelial Cell; Bladder Urothelium; Carbachol; Carcinoma; Cell Membrane Structures; Cell physiology; Cells; Cellular Structures; Cellular biology; Clinical; Complementary DNA; DTR gene; Disease; EGF gene; Electron Microscopy; Epithelial; Epithelial Cells; Esthesia; Freezing; Frequencies; Functional disorder; Goals; Grant; Growth Factor; Health; Herpes zoster disease; Human; Immunofluorescence Immunologic; Immunohistochemistry; In Vitro; Interstitial Cystitis; Ion Channel; Lasers; Lead; Literature; Lower urinary tract; Measures; Mediating; Membrane; Membrane Structure and Function; Messenger RNA; Molecular; Muscarinic M2 Receptor; NIH Program Announcements; Nocturia; Organ; Overactive Bladder; Pain; Pathogenesis; Patients; Plasmids; Play; Potassium; Proteins; Quality of life; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Scanning; Scanning Electron Microscopy; Sensory; Side; Signal Transduction; Specimen; State Interests; Structure; Symptoms; Techniques; Testing; Tight Junctions; Time; Transcript; Transfection; Treatment Factor; Urge Incontinence; Urothelial Cell; Urothelium; Western Blotting; base; design; human disease; improved; large-conductance calcium-activated potassium channels; laser capture microdissection; meetings; micturition urgency; neoplastic; occludin; programs; protein expression; research study; response; transmission process

DESCRIPTION (provided by applicant): The relevance of bladder urothelial cells (BUG) to human disease has traditionally been understood only in terms of urothelial cell carcinoma. However, there is exciting new literature highlighting the role of altered BUG structure and function in non-neoplastic states. Two highly prevalent yet poorly understood conditions, interstitial cystitis (IC) and overactive bladder (OAB) may both be associated with pathogenic changes in the bladder urothelium. Both of these conditions are characterized by hypersensory symptoms such as urinary urgency, frequency, and bladder pain. We propose to study the structure and function of the bladder urothelium as it relates to both normal and hypersensory conditions of IC and OAB. Recent scanning electron microscopy of the lumenal side of the bladder urothelium show distinct changes between IC and control apical umbrella cells (AUC). These changes could be due to abnormalities in uroplakins expression. Uroplakins are unique proteins expressed only by the AUC in the urothelium and play a role in AUC membrane structure and function. Human IC BUC grown in culture also had abnormalities including augmented purinergic signaling, altered expression of certain growth factors (HB-EGF, EGF and APF), and decreased potassium conductance (mediated by ion channels Kir2.1 and BK) leading to depolarization. The discovery of muscarinic M2 receptors on the bladder urothelium provides another mechanism of action in the use of antimuscarinics in treatment of OAB. This grant will test the following 4 hypotheses: 1. AUC from patients with bladder hypersensory conditions have structural abnormalities related to uroplakins. 2. The bladder urothelium from patients with bladder hypersensory conditions have altered protein expressions of tight junction proteins (ZO-1, occludin), Kir2.1, BK, M2, HB-EGF, EGF and APF. 3. The conductance of ion channel Kir2.1 is decreased in IC BUC. This can be reversed with HB-EGF treatment. Normal BUC can be made to have decreased Kir2.1 conductance with APF treatment. 4. The conductance of the BK channel is regulated by M2 receptors in human BUC. There is a decreased outward potassium current in OAB and IC BUC which can be reversed by hSIo cDNA transfection. By accomplishing these aims, we hope to develop an understanding of the pathogenesis of hypersensory bladder conditions which ultimately can lead to targeted treatment options.

描述（由申请人提供）：传统上仅在尿路上皮细胞癌方面了解膀胱尿路上皮细胞（BUG）与人类疾病的相关性。然而，有一些令人兴奋的新文献强调了 BUG 结构和功能改变在非肿瘤状态中的作用。间质性膀胱炎（IC）和膀胱过度活动症（OAB）这两种非常普遍但人们知之甚少的疾病可能都与膀胱尿道上皮的致病性变化有关。这两种情况的特征都是感觉过敏症状，如尿急、尿频和膀胱疼痛。 我们建议研究膀胱尿路上皮的结构和功能，因为它与 IC 和 OAB 的正常和感觉过度状态有关。最近膀胱尿路上皮管腔侧的扫描电子显微镜显示 IC 和对照顶端伞细胞 (AUC) 之间存在明显变化。这些变化可能是由于尿斑蛋白表达异常所致。尿斑蛋白是仅由尿路上皮中的 AUC 表达的独特蛋白质，在 AUC 膜结构和功能中发挥作用。在培养物中生长的人 IC BUC 也存在异常，包括嘌呤能信号增强、某些生长因子（HB-EGF、EGF 和 APF）表达改变以及导致去极化的钾电导降低（由离子通道 Kir2.1 和 BK 介导）。膀胱尿道上皮毒蕈碱 M2 受体的发现为使用抗毒蕈碱药治疗 OAB 提供了另一种作用机制。 该资助将检验以下 4 个假设： 1. 膀胱感觉过敏患者的 AUC 具有与尿斑蛋白相关的结构异常。 2. 膀胱感觉过敏患者的膀胱尿路上皮紧密连接蛋白（ZO-1、occludin）、Kir2.1、BK、M2、HB-EGF、EGF 和 APF 的蛋白表达发生改变。 3. IC BUC中离子通道Kir2.1的电导降低。这可以通过 HB-EGF 治疗来逆转。通过 APF 治疗，可以使正常 BUC 的 Kir2.1 电导降低。 4. BK通道的电导受人BUC中M2受体的调节。 OAB 和 IC BUC 中的外向钾电流减少，这可以通过 hSIo cDNA 转染来逆转。通过实现这些目标，我们希望加深对膀胱感觉过敏疾病发病机制的了解，最终可以制定有针对性的治疗方案。