Role of mechanotransducer Piezo1 in laryngeal and vocal fold epithelial function

机械传感器 Piezo1 在喉和声带上皮功能中的作用

• 批准号: 10064876
• 负责人: Alexander G Foote
• 金额: $ 3.91万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064876
• 关键词: Ablation; Adult; Affect; Alleles; Anatomy; Apical; Apoptosis; Architecture; Biological; Biomechanics; Bladder; Cartilage; Cell Differentiation process; Cell membrane; Cells; Cellular Mechanotransduction; Clinical; Clinical Treatment; Communication impairment; Coughing; Deglutition Disorders; Development; Disease; Embryo; Enterobacteria phage P1 Cre recombinase; Environment; Epithelial; Epithelial Cells; Epithelium; Etiology; Event; Exposure to; Friction; Gastrointestinal tract structure; Generations; Genes; Genetic; Health; Heart; Histology; Homeostasis; Immunofluorescence Immunologic; In Situ Nick-End Labeling; Individual; Inflammatory; Integral Membrane Protein; Ion Channel; Keratin; Knock-out; Knowledge; Laryngeal Diseases; Laryngeal Epithelium; Larynx; Liquid substance; Lung; Maintenance; Mammalian Cell; Mechanical Stress; Mechanics; Mechanoreceptors; Mediating; Mediator of activation protein; Modeling; Molecular; Morphogenesis; Morphology; Mucous Membrane; Mus; Nuclear; Occupational; Outcome; Pathologic Processes; Pathology; Phenotype; Phonation; Physiological Processes; Piezo 1 ion channel; Piezo 2 ion channel; Piezo ion channels; Play; Positioning Attribute; Process; Production; Productivity; Proteins; Regenerative Medicine; Regulation; Respiratory System; Role; Sensory; Sensory Disorders; Series; Signal Pathway; Signal Transduction; Skin; Squamous Epithelium; Stains; Stimulus; Stratified Epithelium; Stratified Squamous Epithelium; Stretching; Structure; Tamoxifen; Testing; Tight Junctions; Tissue Engineering; Tissues; Transducers; Transgenic Organisms; Trauma; Voice Disorders; Work; airway epithelium; base; body system; cell type; cohort; conditional knockout; experimental study; genomic locus; in vitro Model; in vivo; in vivo regeneration; innovation; insight; mechanical force; mechanical load; mechanotransduction; mouse model; mutant; novel; offspring; perinatal development; postnatal; progenitor; promoter; protein biomarkers; protein expression; protein function; regenerative; response; self-renewal; shear stress; somatosensory; stem cells; vocal cord

PROJECT ABSTRACT Voice disorders are among the most common communication disorders and are often associated with mucosa inflammatory etiologies. Furthermore, laryngeal sensory disorders represent a significant clinical cohort that present ubiquitous treatment challenges. Recent discovery of mechanosensitive ion channels, Piezo1 and Piezo2, has proved essential to development and homeostasis for various tissue phenotypes that undergo mechanical loading (e.g. lung, skin, cartilage, bladder, vasculature, heart). As molecular transducers of mechanical force, Piezo proteins are activated by mechanical stretch stimuli exerted on cell membranes, upon which they rapidly and efficiently convert stimuli into intracellular signaling events for biologic effect. Evidence supports Piezo1 presence and function in epithelial cells, however, the role of Piezo1 in vocal fold (VF) – a tissue type that undergoes extreme biomechanical challenges, is not known. In this application, a series of experiments to determine the contribution of Piezo1 to vocal fold epithelium development and function is proposed. We will employ two different in vivo strategies to delete/knockout (KO) Piezo1 using cell specific carriers. Aim 1 will define Piezo1 regulation in vocal fold epithelium morphogenesis and structural barrier protection mechanisms following widespread Piezo1 KO in VF mucosa. Aim 2 will determine Piezo1 function localized to VF basal epithelial cells via Keratin 14 promoter to elucidate differentiation and self-renewal strategies. Our overarching hypothesis is that Piezo1 is a critical mechanotransducer for VF epithelium; essential for a matured, stratified tissue architecture with necessary barrier function. This innovative application will provide vital knowledge for mechanism(s) that govern cell signaling pathways involved in mechanosensory transduction processes in the developing and adult VF epithelium.

项目摘要 语音障碍是最常见的沟通障碍之一，通常与粘膜有关。 炎症病因学。此外，喉感觉障碍代表了一个重要的临床队列， 存在普遍存在的治疗挑战。机械敏感离子通道Piezo 1和Piezo 2的最新发现 Piezo 2，已被证明是必不可少的发展和稳态的各种组织表型，经历 机械负载（例如肺、皮肤、软骨、膀胱、血管系统、心脏）。作为分子传感器， 机械力，压电蛋白被施加在细胞膜上的机械拉伸刺激激活， 它们快速有效地将刺激转化为细胞内信号事件以产生生物学效应。证据 支持Piezo 1在上皮细胞中的存在和功能，然而，Piezo 1在声带（VF）中的作用- a 经受极端生物力学挑战的组织类型是未知的。在本申请中， 确定Piezo 1对声带上皮发育和功能的贡献的实验是 提出了我们将采用两种不同的体内策略，使用细胞特异性引物缺失/敲除（KO）Piezo 1。 载波目的1明确Piezo 1在声带上皮形态发生和结构屏障中的调控作用 VF粘膜中广泛Piezo 1 KO后的保护机制。目标2将确定Piezo 1功能 通过角蛋白14启动子定位于VF基底上皮细胞，以阐明分化和自我更新 战略布局我们的总体假设是，Piezo 1是VF上皮的关键机械换能器; 对于具有必要屏障功能的成熟分层组织结构至关重要。这种创新的应用 将提供重要的知识机制，管理细胞信号通路参与机械感觉 在发育和成年VF上皮中的转导过程。