Identification and activation mechanisms of vagal and spinal nociceptors in esophageal mucosa

食管粘膜迷走神经和脊髓伤害感受器的识别和激活机制

基本信息

批准号：
9978776
负责人：
Thomas Edward Taylor-Clark
金额：
$ 35.24万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-25 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9978776
关键词：
ASIC channel Acids Address Anatomy Animal Model Animals Bile Acids C Fiber Cations Cavia Chest Pain Clinical Data Development Electrophysiology (science)Embryo Esophageal mucous membrane Esophagus Esthesia Failure Family Fiber Functional disorder Future GPBAR1 gene Gastric Acid Gastroesophageal reflux disease Gene Expression Profiling Gene Silencing Genes Genetic Green Fluorescent Proteins Health Heartburn Human Investigation Knowledge Lead Location Mediating Medical Methods Molecular Mucous Membrane Nerve Nerve Fibers Neural Crest Neurons Neurotransmitters Nociception Nociceptors Organ Donor Pain Paper Patients Pharmaceutical Preparations Pharmacology Population Positioning Attribute Potassium Process Proton Pump Inhibitors Protons Publishing Reflex action Reflux Refractory Spinal Stomach Content Symptoms TRPV1 gene Target Populations Therapeutic Tissues United States afferent nerve base differential expression experience in vivo knock-down multiphoton imaging neuroregulation novel pain sensation patch clamp patient population persistent symptom receptor response small hairpin RNA targeted treatment transgene expression

项目摘要

Project Summary Gastroesophageal reflux disease (GERD) is a major health problem in the United States, impacting over 20% of the population. In at least a quarter of these patients, symptoms caused by GERD such as heartburn and chest pain cannot be controlled by gastric acid suppression with proton pumps inhibitors (PPIs). This population of patients with PPI-refractory GERD have no effective medical treatment options. Failure to understand mechanisms of persistent symptoms in this condition is a major knowledge gap, and limits clinical therapeutic options. One potential cause of persistent symptoms in PPI-refractory GERD patients is weakly acidic (pH>5.0) reflux. This can cause activation of pain- mediating nerves (C-fiber nociceptors) in esophageal mucosa, but the molecular mechanisms have not been clearly elucidated. Furthermore it is not clear whether bile acid also directly activates esophageal C-fibers and mediates nociceptive symptoms in GERD. Our overall hypothesis is that the refluxates in PPI-refractory GERD (mild acid and bile acids) activate esophageal nociceptive C-fiber subtypes and contribute to persistent symptoms in refractory GERD. Determining the identity of the afferent nerve subtype(s) and the key receptor(s) responsible for PPI refractory symptoms are crucial for development of specific targets for this population. Over the past decade, we have identified 3 types of C-fibers in the esophagus which express differential receptors and neurotransmitters and have different central projections. Intriguingly, we have developed new data which demonstrated that two factors in weakly acidic refluxate robustly stimulate esophageal C-fibers: 1) bile acids (that are quite often found in the refluxates on PPI therapy), and 2) acid in low proton concentrations (pH=6.0, termed here mild acid). Our published papers and preliminary data suggest that the key receptors TGR5 and TASK1/ASIC3 are likely involved. Based on these progresses, we will first determine in aim 1 the subtypes of nociceptive afferent nerve terminals in esophagus mucosa which are more vulnerable to refluxates. We will then elucidate receptors mediating C-fiber activation by mild acid in aim 2. We will address the hypothesis that mild acid activates C-fiber subtypes via combination of inhibition of potassium two-pore domain (K2P) family channel (TASK1) and activation of cationic channels from the ASIC family (ASIC3). In aim 3, we will elucidate receptors mediating C-fiber activation by bile acids. We hypothesize that bile acids activate C- fiber subtype(s), which is mediated by the bile acid receptor TGR5. We will apply the approaches of gene expression analysis, patch clamp recordings, and recordings of nerve activity originating in the C-fiber nerve terminals. Studies will be carried out in in animal models in which neuron-selective gene knockdown by our validated in vivo shRNA silencing strategy. Elucidation of the mechanisms by which bile acids and mild acid activate C-fibers will help to develop novel targets for PPI--refractory GERD.

项目摘要胃食管反流病（GERD）是美国的主要健康问题，影响了 20％的人口。在至少四分之一的患者中，由GERD引起的症状，例如用质子泵抑制剂抑制胃酸，无法控制胃灼热和胸痛（PPI）。 PP-RECRACTORY GERD的患者人群没有有效的医疗选项。在这种情况下，未能理解持续症状的机制是主要的知识差距，并限制临床治疗选择。持续症状的一个潜在原因 PP-RACTARCTORY GERD患者是酸性弱（pH> 5.0）的反流。这会导致疼痛激活 - 食管粘膜中介导神经（C纤维伤害感受器），但分子机制尚未清楚地阐明了。此外，尚不清楚胆汁酸是否也直接激活食管 C纤维和介导GERD的伤害感受性症状。我们的总体假设是回流 PPI饮食GERD（温和酸和胆汁酸）激活食道伤害性C纤维亚型，并激活有助于难治性GERD的持续症状。确定传入神经的身份负责PPI难治症状的亚型和关键受体对于发育至关重要该人群的特定目标。在过去的十年中，我们已经确定了3种类型的C纤维表达差异受体和神经递质的食道，具有不同的中心预测。有趣的是，我们开发了新数据，这些数据表明弱的两个因素酸性回流可靠地刺激食管C纤维：1）胆汁酸（通常在 PPI疗法的回流）和2）低质子浓度（pH = 6.0，称为温和酸）的酸。我们的已发表的论文和初步数据表明，关键受体TGR5和TEAKS1/ASIC3可能是涉及。基于这些进展，我们将首先在AIM 1中确定伤害性传入的亚型食管粘膜中的神经末端更容易流动。然后我们将阐明在AIM 2中介导C纤维激活C纤维激活的受体。我们将解决温和酸的假设通过组合抑制钾两孔结构域（K2P）家族来激活C纤维亚型通道（任务1）和来自ASIC家族的阳离子通道的激活（ASIC3）。在AIM 3中，我们将阐明胆汁酸介导C纤维激活的受体。我们假设胆汁酸激活C- 纤维亚型（S），由胆汁酸受体TGR5介导。我们将应用基因的方法表达分析，斑块夹记录以及源自C纤维中的神经活动的记录神经终端。研究将在神经元选择基因的动物模型中进行通过我们经过验证的体内shRNA沉默策略敲除。阐明该机制胆汁酸和温和酸激活的C纤维将有助于开发PPI - 残酷GERD的新目标。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Acidic Pharyngeal Reflux Does Not Correlate with Symptoms and Laryngeal Injury Attributed to Laryngopharyngeal Reflux.

DOI：
10.1007/s10620-018-5372-1
发表时间：
2019-05
期刊：
DIGESTIVE DISEASES AND SCIENCES
影响因子：
3.1
作者：
Duricek, Martin;Banovcin, Peter;Halickova, Tatiana;Hyrdel, Rudolf;Kollarik, Marian
通讯作者：
Kollarik, Marian

Comprehensive analysis of acidic pharyngeal reflux before and after proton pump inhibitor treatment in patients with suspected laryngopharyngeal reflux.

DOI：
10.1097/meg.0000000000001584
发表时间：
2020-02
期刊：
EUROPEAN JOURNAL OF GASTROENTEROLOGY & HEPATOLOGY
影响因子：
2.1
作者：
Duricek, Martin;Banovcin, Peter, Jr.;Halickova, Tatiana;Hyrdel, Rudolf;Kollarik, Marian
通讯作者：
Kollarik, Marian

Mapping of the Sensory Innervation of the Mouse Lung by Specific Vagal and Dorsal Root Ganglion Neuronal Subsets.

通过特定迷走神经和背根神经节神经元亚群绘制小鼠肺的感觉神经支配图。

DOI：
10.1523/eneuro.0026-22.2022
发表时间：
2022
期刊：
eNeuro
影响因子：
3.4
作者：
Kim,Seol-Hee;Patil,MayurJ;Hadley,StephenH;Bahia,ParmvirK;Butler,ShaneG;Madaram,Meghana;Taylor-Clark,ThomasE
通讯作者：
Taylor-Clark,ThomasE

Structure of vagal afferent nerve terminal fibers in the mouse trachea.

DOI：
10.1016/j.resp.2018.01.001
发表时间：
2018-03
期刊：
Respiratory physiology & neurobiology
影响因子：
2.3
作者：
Hennel M;Harsanyiova J;Ru F;Zatko T;Brozmanova M;Trancikova A;Tatar M;Kollarik M
通讯作者：
Kollarik M

Ginger Constituent 6-Shogaol Attenuates Vincristine-Induced Activation of Mouse Gastroesophageal Vagal Afferent C-Fibers.

DOI：
10.3390/molecules27217465
发表时间：
2022-11-02
期刊：
MOLECULES
影响因子：
4.6
作者：
Patil, Mayur J.;Huang, Yongming;Yu, Mingwei;Dong, Xinzhong;Undem, Bradley J.;Yu, Shaoyong
通讯作者：
Yu, Shaoyong