Purinergic Regulation of Bladder Interstitial Cells of Cajal

Cajal 膀胱间质细胞的嘌呤能调节

基本信息

批准号：
8707443
负责人：
weiqun yu
金额：
$ 9万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8707443
关键词：
ATP Receptors ATP phosphohydrolase Acetylcholine Adenosine Agonist Animal Model Birth Bladder Bladder Control Bladder Diseases Calcium Calcium Signaling Cells Co-Immunoprecipitations Communication Connexin 43 Connexins Coupled Cyclic AMP Data Disease Enzymes Etiology Exhibits Functional disorder Gap Junctions Genetically Modified Animals Goals Image Incontinence Infant Interstitial Cell of Cajal Interstitial Cystitis Intestines Investigation Kinetics Knowledge Life Manuscripts Mass Spectrum Analysis Mediating Mentors Methods Modeling Molecular Molecular Target Motor Mus Muscle Muscle Contraction Myography Nerve Fibers Neurogenic Bladder Neurons Neurotransmitters Overactive Bladder Pain Pathway interactions Peristalsis Phase Play Proto-Oncogene Protein c-kit Publications Publishing Purinergic P1 Receptors Purinoceptor Receptor Activation Regulation Relaxation Research Role Ryanodine Receptor Calcium Release Channel Signal Pathway Signal Transduction Smooth Muscle Smooth Muscle Myocytes Solutions Staging Symptoms Syndrome Testing Therapeutic Time Tissues Urinary Incontinence Urology Woman Work aging population base cell motility cell type cost desensitization gastrointestinal improved innovation liquid chromatography mass spectrometry lower urinary tract symptoms molecular marker nodal myocyte novel prevent public health relevance receptor receptor expression receptor function research study stoichiometry tool transmission process treatment strategy tripolyphosphate

项目摘要

DESCRIPTION (provided by applicant): Urinary incontinence, overactive bladder (OAB) and neurogenic bladder often arise from inappropriate bladder smooth muscle (BSM) motility and the underlying mechanisms are poorly understood. Pacemaker cells, also known as interstitial cells of Cajal (ICC) are likely to play a critical role in modulating bladder smooth muscle functio but this recently defined cell, is almost completely unstudied. Infants born without bladder ICC (BICC) - a lethal condition called megacystis-microcolon intestinal hypoperistalsis syndrome (MMIHS) - have a complete absence of autonomic voiding function and die with dilated bladders, underscoring a vital role for BICC in modulating BSM. The long-term goal of this research, in alignment with several stated goals of the National Urology Research Agenda, is to fully understand the interactions of neurons, BICC, and BSM in regulating bladder motility. The objective in this particular application is to identify the purinergic signaling pathways which operate in BICC and how they function to regulate BSM motility. We hypothesize that purinergic signaling to BICC will regulate BSM motility through calcium signaling and gap junction transmission. Guided by strong preliminary data demonstrating novel purinergic receptors, P2X2/6 and A2a expression on BICC and observing that activation of these receptors induces BSM contraction/relaxation, we will investigate our hypothesis through the following four specific aims: 1) to demonstrate that BICC mediates BSM contraction through activation of P2X2/6 heteromeric receptors; 2) to define whether P2X2/6 activation results in Ca2+ release from ER and mediates BSM contraction through gap junction transmission; 3) to define whether activation of A2a receptors relaxes BICC-mediated BSM contraction through inhibiting Ca2+ signaling; and 4) to determine whether ectonucleotidase Entpd2 regulates P2X2/6 and A2a receptor function and bladder motility through modulating the availability of ATP and adenosine on BICC. Due to the compelling preliminary data which defines the feasibility of this plan, we expect Aim 1 to be completed during the mentored phase of the project and the resulting manuscript prepared and submitted for publication. Preliminary experiments in pursuit of aims 2 and 3 will likely have begun in this initial phase also, thereby generating important momentum for completion of the remaining aims during the independent research phase. The approach we propose is innovative and integrative. To achieve our aims we have established a new method to study BICC-mediated BSM motility. We will use multiple genetically modified animal models to investigate the involvement of various signaling pathways using bladder muscle strip myography in conjunction with specific pharmacological modulation of receptors, real-time calcium imaging and mass spectrometry. We expect this research to vertically advance our understanding of how BSM motility is regulated by BICC, and eventually establish a novel BICC-BSM interaction model that will shift the paradigm for how bladder motility is regulated. Ultimately, this work may define new treatment solutions and molecular targets for bladder disease.

描述（由申请人提供）：尿失禁、膀胱过度活动症（OAB）和神经源性膀胱通常由不适当的膀胱平滑肌（BSM）运动引起，并且其潜在机制知之甚少。起搏细胞，也称为卡哈尔间质细胞 (ICC)，可能在调节膀胱平滑肌功能中发挥关键作用，但这种最近定义的细胞几乎完全未经研究。出生时没有膀胱 ICC (BICC) 的婴儿 - 一种称为巨囊小结肠肠蠕动功能减退综合征 (MMIHS) 的致命疾病 - 完全缺乏自主排尿功能，并因膀胱扩张而死亡，这强调了 BICC 在调节 BSM 中的重要作用。这项研究的长期目标是充分了解神经元、BICC 和 BSM 在调节膀胱运动方面的相互作用，这与国家泌尿学研究议程的几个既定目标保持一致。该特定应用的目的是确定 BICC 中起作用的嘌呤能信号通路以及它们如何发挥调节 BSM 运动的作用。我们假设 BICC 的嘌呤能信号传导将通过钙信号传导和间隙连接传输调节 BSM 运动。以强有力的初步数据为指导，证明 BICC 上存在新型嘌呤能受体、P2X2/6 和 A2a 表达，并观察到这些受体的激活会诱导 BSM 收缩/松弛，我们将通过以下四个具体目标来研究我们的假设：1）证明 BICC 介导通过激活 P2X2/6 异聚受体来收缩 BSM； 2) 确定P2X2/6激活是否导致Ca2+从ER释放并通过间隙连接传递介导BSM收缩； 3) 确定A2a受体的激活是否通过抑制Ca2+信号传导来放松BICC介导的BSM收缩； 4) 确定核酸外切酶Entpd2是否通过调节BICC上ATP和腺苷的可用性来调节P2X2/6和A2a受体功能和膀胱运动。由于确定该计划可行性的令人信服的初步数据，我们预计目标 1 将在项目的指导阶段完成，并准备好并提交出版的手稿。追求目标 2 和 3 的初步实验也可能在这个初始阶段开始，从而为独立研究阶段完成其余目标产生重要动力。我们提出的方法是创新和综合的。为了实现我们的目标，我们建立了一种新方法来研究 BICC 介导的 BSM 运动。我们将使用多种转基因动物模型，利用膀胱肌条肌描记术结合受体的特定药理调节、实时钙成像和质谱来研究各种信号传导途径的参与。我们期望这项研究能够纵向推进我们对 BICC 如何调节 BSM 运动的理解，并最终建立一种新颖的 BICC-BSM 相互作用模型，该模型将改变膀胱运动调节的范式。最终，这项工作可能会为膀胱疾病定义新的治疗方案和分子靶点。