Role of Ectonucleotidase in Voiding Function and Dysfunction

外切核苷酸酶在排尿功能和功能障碍中的作用

• 批准号: 10292995
• 负责人: weiqun yu
• 金额: $ 38.5万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-19 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10292995
• 关键词: ATP Hydrolysis; Address; Adenosine; Adrenergic Agents; Affect; Age; Aging; Agonist; Appearance; Atropine; Autoimmune; Autoimmune Diseases; Bladder; Bladder Control; Bladder Diseases; Bladder Dysfunction; Bladder Tissue; Clinical; Comparative Study; Cost of Illness; Data; Diabetes Mellitus; Disease; Drug Targeting; Enzymes; Exhibits; Functional disorder; Genes; Human; Impairment; Kinetics; Knock-out; Knockout Mice; Knowledge; Light; LoxP-flanked allele; Mediating; Mediator of activation protein; Molecular; Mouse Strains; Mus; Muscarinic Antagonists; Muscarinics; Mutation; Myocardium; Nature; Nerve; Neurodegenerative Disorders; Overactive Bladder; Partner in relationship; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Physiology; Play; Population; Purinoceptor; Receptor Signaling; Regulation; Relaxation; Reporting; Research; Rodent; Role; Seeds; Signal Pathway; Signal Transduction; Smooth Muscle; Smooth Muscle Myocytes; Stimulus; Symptoms; Testing; Therapeutic; Time; Tissues; Transgenic Mice; Translating; Urination; Visceral; cost; design; diabetic; differential expression; disabling symptom; health related quality of life; human disease; human female; insight; loss of function mutation; lower urinary tract symptoms; male; mouse model; mutant mouse model; new therapeutic target; novel; novel therapeutic intervention; novel therapeutics; prevent; receptor; receptor expression; receptor function; sex; social; telokin

Abstract: Lower urinary tract symptoms (LUTS) impact a large percentage of the population with enormous financial and social costs, and effective drugs for management of LUTS are limited. Purinergic signaling is a prominent dysregulated pathway, which has been consistently reported in LUTS. While purinergic contractility is minimal in normal human bladder, it increases significantly in bladders of human patients, to more closely resembles documented rodent physiology. However, the underlying mechanism which results in these functional pathway alterations is not known. To study this, we have identified multiple novel functional purinergic receptors and modulators in bladder. We have demonstrated that purinergic signaling is not only important for contraction in bladder wall, as mediated by ATP/ADP activated P2X1/P2Y12 signaling, but is also important for bladder relaxation mediated by adenosine activated A2b receptor signaling. Furthermore, A2b receptor mediated relaxation signaling can completely inhibit P2X1/P2Y12 mediated contractions. By comparative studies, we have defined a key gene - ENTPD1 (also known as CD39) – that is differentially expressed in human and mouse bladders (~ 8-fold higher in normal human bladders than in mouse). ENTPD1 is a rate-limiting enzyme converting ATP/ADP/AMP sequentially. ENTPD1 controls the kinetics of an agonist cascade (ATP>ADP>adenosine), and hence plays a critical role in the control of purinergic receptor expression and function. Interestingly, human ENTPD1 mutations have been identified in autoimmune diseases and neurodegenerative disorders, and LUTS is a prominent symptom in these diseases. Consistent with this, impaired ATP hydrolysis has been reported in bladder tissues from LUTS patients. Therefore, we hypothesize that ENTPD1 is a key regulator in bladder function. In support, we have preliminary data from a global Entpd1- /- mouse, indicating that deletion of Entpd1 results in a dilated bladder with abnormal voiding phenotype. We will test our hypothesis through the following aims: (1) We will generate a visceral smooth muscle specific Entpd1 knockout mouse model to demonstrate the critical importance of ENTPD1 in regulating normal detrusor contractility and bladder function; (2) We will use two ENTPD1 mutant mouse models with high and low ENTPD1 expression levels to mimic human bladders that are normal (high) and diseased (low), and then examine voiding function/dysfunction; (3) Through the National Disease Research Interchange (NDRI) network, we will procure human bladder tissues, and study the expression and function of ENTPD1 in the bladders of both sexes. We will solve the puzzles of why there are purinergic contractile differences between human and rodent bladders, between male and female human bladders, and between normal and diseased human bladders. We expect this study to substantially advance our understanding on how bladder contraction and relaxation is controlled by ENTPD1 and its associated purinergic signaling. These studies have high potential to introduce novel therapies for human LUTS patients as there are multiple targetable options.

抽象的： 较低的尿路症状（LUTS）影响了很大一部分的财务和 社会成本和用于LUTS管理的有效药物是有限的。嘌呤能信号传导是突出的 失调的途径，在LUTS中一直报告。而嘌呤能收缩性很小 在正常的人膀胱中，人类患者的膀胱大大增加，与更相似 记录的啮齿动物生理学。但是，导致这些功能途径的基本机制 改变是不知道的。为了研究这一点，我们已经确定了多个新型的功能性嘌呤能受体和 膀胱调节器。我们已经证明，嘌呤能信号不仅对收缩很重要 膀胱壁，由ATP/ADP激活的P2X1/P2Y12信号传导介导，但对于膀胱也很重要 腺苷介导的弛豫激活了A2B受体信号。此外，A2B受体介导 松弛信号传导可以完全抑制P2X1/P2Y12介导的收缩。通过比较研究，我们有 定义了一个关键基因-ENTPD1（也称为CD39），该基因在人和小鼠中的表达不同 膀胱（正常人膀胱中比小鼠高约8倍）。 ENTPD1是限速酶 顺序转换ATP/ADP/AMP。 ENTPD1控制激动剂级联的动力学 （ATP> ADP>腺苷），因此在控制嘌呤能受体表达和 功能。有趣的是，在自身免疫性疾病中已经确定了人ENTPD1突变 神经退行性疾病和LUTS是这些疾病的重要症状。与此一致 LUTS患者的膀胱组织中已经报道了ATP水解受损。因此，我们假设 该ENTPD1是膀胱功能中的关键调节器。为了支持，我们拥有来自全球ENTPD1-的初步数据 / - 小鼠，表明ENTPD1的缺失导致膀胱扩张，异常排尿表型。我们 将通过以下目的检验我们的假设：（1）我们将产生一个内脏平滑肌特异性 ENTPD1敲除鼠标模型，以证明ENTPD1在控制正常急躁方面的重要性 收缩和膀胱功能； （2）我们将使用两个ENTPD1突变小鼠模型高和低 ENTPD1表达水平与模仿正常（高）和患病（低）的人类膀胱，然后 检查空隙功能/功能障碍； （3）通过国家疾病研究互换（NDRI） 网络，我们将采购人类膀胱组织，并研究ENTPD1在 两性的膀胱。我们将解决为什么嘌呤能收缩之间存在差异的难题 男性和雌性人类膀胱之间以及正常和衰落之间的人类和啮齿动物膀胱 人类的刀具。我们希望这项研究能够大大提高我们对膀胱收缩的理解 放松由ENTPD1及其相关的嘌呤能信号控制。这些研究很高 由于有多种目标选择，因此潜力为人类LUTS患者引入新型疗法。