Nanoscale structure and function of desmosomes

桥粒的纳米结构和功能

基本信息

批准号：
10380815
负责人：
Alexa Lynn Mattheyses
金额：
$ 32.34万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
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：
2018-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10380815
关键词：
Address Adherens Junction Adhesions Adhesives Antibodies Architecture Autoantibodies Binding Biological Markers Biophysics Biopsy Specimen Bulla C-terminal Cadherin Domain Cadherins Calcium Cell Adhesion Cell-Cell Adhesion Cells Cellular biology Characteristics Complex Core Protein Couples Cytoskeleton Defect Dehydration Dependence Dermatologic Desmosomes Development Disease Electrons Elements Environmental Protection Epidermis Epithelial Fluorescence Microscopy Future Goals Growth Heart Diseases Human Image Immunoglobulin G Individual Intermediate Filaments Lead Life Link Macromolecular Complexes Maps Measures Mechanical Stress Mechanics Mediating Methods Microscopy Modeling Molecular Multiprotein Complexes Optics Organizational Change PRKCA gene Pathogenesis Pemphigus Vulgaris Phosphorylation Polarization Microscopy Primary Cell Cultures Process Protein Dynamics Proteins Regulation Resistance Resolution Signal Transduction Skin Stress Structure Structure-Activity Relationship Tertiary Protein Structure Testing Tissue Sample Tissues desmoglein III desmoplakin experimental study extracellular human disease human tissue innovation insight interdisciplinary approach keratinocyte mutant nanoscale novel novel strategies potential biomarker skin disorder targeted treatment therapeutic development therapeutic target tool wound healing

项目摘要

Project Summary The epidermis provides protection from environmental insult, dehydration and stress. Mechanical strength is derived from robust cell-cell adhesive junctions called desmosomes is a fundamental feature of epidermal tissue. Desmosomes are macromolecular complexes composed of desmosomal cadherins, which mediate cell- cell adhesion, and a number of intracellular plaque proteins, including desmoplakin, which couples the complex to the intermediate filament cytoskeleton. Notably, aberrant desmosome function can lead to severe epidermal disorders. Pemphigus vulgaris is a potentially life-threatening skin blistering disease caused by autoantibodies directed against the desmosomal cadherin desmoglein-3 (Dsg3) that leads to disruption of cell-cell adhesion. Though responsible for mechanical integrity, desmosomes can switch between strong and weak states in development and wound healing. This functional transition occurs with minimal change to the core proteins comprising the desmosome. We hypothesize that the architecture or organization of proteins within a desmosome drives its adhesive function. However, due to the size and molecular complexity of desmosomes there is a lack of tools to study this structure-function relationship creating a critical barrier in this field. We will use a multi-disciplinary approach to address this challenge and to test the hypothesis that the biophysical organization of proteins in the desmosome provides a mechanism to regulate adhesion. We recently developed two highly innovative and complimentary super-resolution fluorescence microscopy approaches to study the order and organization of proteins within desmosomes. Our goal is to elucidate how the order and organization of proteins impacts the adhesive function of desmosomes in healthy and disease states. This will provide novel insight into the structure and function of these critical complexes. In Aim 1 we will determine the how the organization of plaque proteins changes in different adhesive states with the goal of identifying functionally sensitive elements and potential biomarkers. In Aim 2 we will use a live cell approach to study mechanisms that confer ordering of desmosomal cadherins, and how this order is altered with function. Finally, in Aim 3 we will define changes to the architecture of the desmosome induced in pemphigus vulgaris, with the goal of facilitating development of targeted therapeutics. We will use primary human keratinocytes and human tissue biopsy samples to address these questions. Accomplishment of these goals will provide a fundamental understanding and framework of how protein organization and dynamics influence the adhesive function of desmosomes in healthy and disease states.

项目概要表皮提供保护，免受环境侵害、脱水和压力。机械强度为源自称为桥粒的坚固的细胞与细胞粘合连接，是表皮的基本特征组织。桥粒是由桥粒钙粘蛋白组成的大分子复合物，介导细胞- 细胞粘附和许多细胞内斑块蛋白，包括桥粒斑蛋白，它与复合物偶联到中间丝细胞骨架。值得注意的是，异常的桥粒功能可导致严重的表皮损伤失调。寻常型天疱疮是一种由自身抗体引起的可能危及生命的皮肤起泡疾病针对桥粒钙粘蛋白 desmoglein-3 (Dsg3)，导致细胞间粘附破坏。虽然桥粒负责机械完整性，但它可以在强状态和弱状态之间切换发育和伤口愈合。这种功能转变发生时核心蛋白的变化极小包括桥粒。我们假设蛋白质的结构或组织桥粒驱动其粘附功能。然而，由于桥粒的大小和分子复杂性缺乏研究这种结构-功能关系的工具，在该领域造成了严重障碍。我们将使用多学科方法来应对这一挑战并检验生物物理学的假设桥粒中蛋白质的组织提供了调节粘附的机制。我们最近开发了两种高度创新和互补的超分辨率荧光显微镜方法研究桥粒内蛋白质的顺序和组织。我们的目标是阐明顺序和蛋白质的组织影响健康和疾病状态下桥粒的粘附功能。这将为这些关键复合体的结构和功能提供新的见解。在目标 1 中，我们将确定斑块蛋白的组织在不同的粘附状态下如何变化，目的是识别功能敏感元件和潜在的生物标志物。在目标 2 中，我们将使用活细胞方法进行研究赋予桥粒钙粘蛋白排序的机制，以及这种顺序如何随着功能而改变。最后，在目标 3 中，我们将定义寻常型天疱疮中诱导的桥粒结构的变化，其中促进靶向治疗药物开发的目标。我们将使用原代人角质形成细胞和人组织活检样本可以解决这些问题。这些目标的实现将为蛋白质组织和动力学如何影响粘附功能的理解和框架健康和疾病状态下的桥粒。