Purinergic Regulation of Bladder Interstitial Cells of Cajal

Cajal 膀胱间质细胞的嘌呤能调节

• 批准号: 9143745
• 负责人: weiqun yu
• 金额: $ 24.9万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-11 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9143745
• 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; Muscle function; 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; Staging; Symptoms; Syndrome; Testing; Therapeutic; Time; Tissues; Urinary Incontinence; Urology; Woman; Work; abstracting; 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; receptor; receptor expression; receptor function; research study; stoichiometry; tool; transmission process; treatment strategy; tripolyphosphate

Title: Purinergic Regulation of Bladder Interstitial Cells of Cajal Abstract: 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 function 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.

卡哈尔膀胱间质细胞的嘌呤能调节 摘要：尿失禁、膀胱过度活动(OAB)和神经源性膀胱通常由以下原因引起 不适当的膀胱平滑肌(BSM)运动及其潜在机制尚不清楚。 起搏细胞，也被称为Cajal间质细胞(ICC)，可能在调节 膀胱平滑肌的功能，但这一新近定义的细胞，几乎完全没有研究。出生的婴儿 无膀胱ICC(BICC)--一种称为巨囊炎的致命性疾病--微结肠肠动力低下 综合征(MMIHS)-完全缺乏自主排尿功能，死于膀胱扩张， 强调了BICC在调节BSM方面的重要作用。这项研究的长期目标，与 国家泌尿学研究议程的几个明确目标是充分了解 神经元、BICC和BSM在调节膀胱运动中的作用。此特定应用程序目标是识别 在BICC中工作的嘌呤能信号通路以及它们如何调节BSM的运动。我们 假设BICC的嘌呤能信号将通过钙信号和GAP调节BSM的运动 连接变速箱。在强大的初步数据的指导下，展示了新的嘌呤能受体，P2X2/6和 A2a在BICC上的表达，观察到这些受体的激活诱导了BSM的收缩/松弛， 我们将通过以下四个具体目标来检验我们的假设：1)证明BICC 通过激活P2X2/6异构体受体介导BSM收缩；2)确定P2X2/6是否 激活导致内质网钙释放，并通过缝隙连接传递介导BSM收缩；3) A2a受体激活是否通过抑制钙离子而松弛BICC介导的BSM收缩 信号转导；以及4)确定ECN Entpd2是否调节P2X2/6和A2a受体功能 通过调节三磷酸腺苷和腺苷在BICC上的可获得性来调节膀胱的运动。由于令人信服的 初步数据确定了该计划的可行性，我们预计目标1将在年内完成。 该项目的指导阶段以及编写并提交供出版的最终手稿。 追求目标2和目标3的初步试验也可能在这一初始阶段开始，因此 在独立研究阶段为完成剩余目标创造重要动力。 我们提出的方法是创新的和综合的。为了实现我们的目标，我们建立了一个新的 方法研究BICC介导的BSM运动。我们将使用多种转基因动物模型来 联合应用膀胱肌条状成像研究不同信号通路的参与 具有特异性的受体药理调节，实时钙成像和质谱学。我们 期待这项研究垂直推进我们对BICC如何调节BSM运动性的理解，以及 最终建立一个新的BICC-BSM相互作用模型，该模型将改变膀胱运动的范式 受监管的。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。