Mechanisms of mechanotransduction in the enterochromaffin cells

肠嗜铬细胞中的机械转导机制

• 批准号: 8948535
• 负责人: Arthur Beyder
• 金额: $ 15.34万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-16 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8948535
• 关键词: Absenteeism; Acute; Address; Affect; Animals; Area; Basic Science; Biology; Biophysics; Cartoons; Cations; Cell Line; Cell physiology; Cells; Chronic; Clinic; Colon; Consumption; Coupled; Couples; Cytoskeleton; Data; Development; Direct Expenditure; Disease; Doctor of Philosophy; Electric Capacitance; Electrochemistry; Electrophysiology (science); Elements; Enterochromaffin Cells; Environment; Epithelial; Epithelial Cells; Epithelium; Esthesia; Foundations; Functional Gastrointestinal Disorders; Functional disorder; Gastroenterology; Gastrointestinal Motility; Gastrointestinal tract structure; Goals; Health; Healthcare; Hepatology; Human; Image; Immunohistochemistry; International; Intestines; Ion Channel; Irritable Bowel Syndrome; K-Series Research Career Programs; Knock-out; Life; Link; Manometry; Measurement; Mechanics; Mentors; Mentorship; Merkel Cells; Methods; Microscopy; Molecular; Morbidity - disease rate; Mus; Organoids; Outcome; Physicians; Physiological; Physiology; Population; Postdoctoral Fellow; Productivity; Research; Resolution; Role; Scientist; Serotonin; Signal Transduction; Skin; Stimulus; Symptoms; Techniques; Testing; Therapeutic; Tissues; Training; Training Programs; Transgenic Animals; Visceral; Visceral Afferents; Work; authority; career; cell motility; cell type; cellular targeting; gastrointestinal; gastrointestinal epithelium; in vivo; knock-down; new therapeutic target; novel; novel diagnostics; optogenetics; public health relevance; receptor; research study; skills; targeted treatment; tool

 DESCRIPTION (provided by applicant): This Mentored Career Development Award (K08) proposal describes a five year training program for the physician scientist candidate with a long-term goal of becoming an academic authority in cellular and molecular mechanosensitivity in the gastrointestinal (GI) tract in health and disease. The candidate acquired a strong basic science foundation in ion channel biophysics during his PhD and post-doctoral work. The candidate now proposes to solidify his foundation and acquire a set of novel complementary skills necessary for an independent research career in GI tract mechanosensitivity. Mechanosensitivity is important for normal GI tract function and abnormal mechanosensitivity leads to disease. Enterochromaffin (EC) cells are mechanosensitive GI epithelial cells that synthesize and release serotonin (5-HT), which regulates GI motility and sensation. Mechanical forces are a major stimulus for 5-HT release by the EC cell, but the molecular mechanisms of EC cell mechanosensation are unclear. Acute cellular mechanosensation involves mechanosensitive ion channels (MSCs). A recently cloned MSC PIEZO2 is critical for Merkel cell mechanosensation, which is a mechanosensitive cell of the skin epithelium with developmental and functional similarities to the EC cell. The central hypothesis of this application is that PIEZO2 ion channels are the primary mechanosensors that transduce mechanical energy into 5-HT release by the EC cells. We will test the central hypothesis in 2 AIMs. The experiments in AIM 1 will determine the localization and mechanical activation of PIEZO2 in EC cells. The experiments in AIM 2 will determine how PIEZO2 activation is coupled to 5-HT release in single EC cells and intestinal organoids and how PIEZO2 activation couples to GI motility. The AIMs are supported by strong preliminary data which show that PIEZO2 is specific to human and mouse colon EC cells, that mechanical stimuli activate PIEZO2 in primary EC cells, leading to release of 5-HT, while the block of PIEZO2 in the colon epithelium decreases motility. To test the central hypothesis, we will use immunohistochemistry and super resolution imaging, electrophysiology and optogenetics in parallel with mechanostimulation of single cells and intestinal organoids, 5-HT measurements by electrochemistry and novel techniques, novel ex vivo and in vivo recordings of colonic motility in wild type and transgenic animals. This work will be performed in an academically nurturing environment within Mayo Clinic and with full support of the Division of Gastroenterology & Hepatology. The candidate will be supported by several consultants who are experts in the areas that the candidate identified as necessary training. He will be guided by a strong mentorship committee (Drs. Gianrico Farrugia and Nicholas LaRusso). Both are international authorities - Dr. Farrugia in ion channels in GI motility, and Dr. LaRusso in epithelial biology and mechanosensation. As a result of this work, the candidate will significantly advance our understanding of EC cell mechanosensitivity mechanisms in human physiology with a goal of providing a novel platform for therapeutic strategies as well as obtain data for a R01 application.

 描述（由适用提供）：该指导的职业发展奖（K08）提案描述了针对物理科学家候选人进行的为期五年的培训计划，其长期目标是成为胃肠道（GI）健康和疾病的细胞和分子机制的学术权威。候选人在博士学位和博士后工作中获得了离子频道生物物理学的强大基础科学基金会。现在，提议巩固他的基础，并获得一系列在胃肠道机制独立研究职业所需的新颖的补充技能。机械敏感性对于正常的胃肠道功能很重要，异常机制导致疾病。肠球胶质素（EC）细胞是合成和释放血清素（5-HT）的机械GI上皮细胞，可调节胃肠道运动和感觉。机械力是通过EC细胞释放5-HT的主要刺激，但是EC细胞机制的分子机制尚不清楚。急性细胞机制涉及机理敏感的离子通道（MSC）。最近克隆的MSC Piezo2对于默克尔细胞机制至关重要，这是皮肤上皮的机械敏感细胞，与EC细胞具有发育和功能相似性。该应用的中心假设是我们将在两个目标中测试中心假设。 AIM 1中的实验将确定压电2在EC细胞中的定位和机械激活。 AIM 2中的实验将确定如何在单个EC细胞和肠道类器官中耦合Piezo2激活与5-HT释放，以及如何将Piezo2激活夫妇耦合到GI运动。 AIM 2中的实验将确定如何将Piezo2激活耦合到单个EC细胞和肠道类器官以及Piezo2激活夫妇如何与GI运动性偶联。目标得到了强大的初步数据的支持，这些数据表明，压电是针对人和小鼠结肠EC细胞的，机械刺激激活原代EC细胞中的Piezo2，导致5-HT释放，而结肠上皮的Piezo2块降低了运动性。为了检验中心假设，我们将使用免疫组织化学和超级分辨率成像，电生理学和光遗传学与单细胞和肠癌的机械刺激并联，通过新型技术和新型技术，新型Ex ex ex Vivo以及在野生型和转基因动物中的结肠运动能力中进行的5-HT测量。这项工作将 在梅奥诊所内的学术培养环境中表现出色，并在胃肠病学和肝病学的全面支持下。候选人将得到几位顾问的支持，这些顾问是候选人确定为必要培训领域的专家。他将受到一个强大的心态委员会的指导（Gianrico Farrugia和Nicholas Larusso博士）。两者都是国际当局 - 胃肠道运动的离子频道的Farrugia博士和博士 上皮生物学和机制传感的劳动。这项工作的结果是，候选人将大大提高我们对人类生理学中EC细胞机制的理解，目的是为治疗策略提供新颖的平台，并为R01应用程序获得数据。