The Role of Neutrophil Mechanosensing During Oropharyngeal Candidiasis

中性粒细胞机械感应在口咽念珠菌病中的作用

• 批准号: 10160640
• 负责人: Hadley A. Witt
• 金额: $ 0.58万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10160640
• 关键词: 3-Dimensional; Adhesions; Affect; Antifungal Agents; Biochemical; Biomedical Engineering; Biomimetics; Blood Circulation; Candida; Candidiasis; Cells; Characteristics; Chemicals; Chemotaxis; Chemotherapy-Oncologic Procedure; Denture Stomatitis; Dentures; Development; Devices; Diabetes Mellitus; Disease; Elasticity; Esophagus; Extracellular Matrix; Gel; Generations; Gingiva; Glass; Growth; HIV; Hard Palate; Host Defense; Human; Human body; Hydrogels; Hyphae; Immune; Immunotherapeutic agent; In Vitro; Infection; Inflammation; Innate Immune Response; Integrins; Invaded; Investigation; Laboratories; Lead; Mandible; Mechanics; Mediating; Modality; Motion; Mucous Membrane; Mycoses; Nature; Oral candidiasis; Oral cavity; Organ; Patients; Process; Production; Property; Reporting; Risk; Role; Site; Smoking; Soft Palate; Structure; Surface; System; Testing; Tissues; Tongue; Traction; Variant; Work; antimicrobial; beta-Glucans; cell motility; comorbidity; first responder; immune activation; interstitial; mechanical properties; mechanotransduction; microbial; microorganism; migration; neutrophil; novel therapeutics; oropharyngeal thrush; pathogen; physical property; response; screening; soft tissue; tissue culture

Project Summary / Abstract Neutrophils are the first responders to sites of infection and are fundamental to the anti-fungal response. This includes conditions of oropharyngeal candidiasis, where blood neutrophils extravasate into infected tissues and locomote to contact invading Candida. During this process, neutrophils are highly sensitive not only to the biochemical properties of the tissue microenvironment but the physical properties as well. The oral cavity is highly variant in the mechanical properties of its composite tissues, from the relatively soft and elastic tongue and soft palate to structures of relatively intermediate stiffness such as the esophagus to rigid structures such as the hard palate. The Reichner lab and others have shown that human neutrophils are highly sensitive to the mechanical features of their microenvironment including substrate stiffness. They have reported that human neutrophils move more slowly and with greater directionality when placed on stiff matrices as compared to soft matrices. Quantification and mapping of tractional forces show greater force generation on stiffer matrices with most of the force produced at the rear of the cell. A significant gap regarding these findings is that we have no information regarding how a pathogen, or its component PAMPs, affect the mechanosensitive response of neutrophils with regard to overall motility and the production of traction on matrices of varying stiffnesses. This is significant because it is the pathogen that induces neutrophil entry into an infected tissue to execute antimicrobial activity, and Candida infection is highly relevant in the oral cavity as a cause of OPC. The overarching hypothesis to be tested in this proposal is that the presence of candida affects the mechanosensitive properties of the human neutrophil. In Aim 1, we will use tunable elasticity hydrogels to determine how substrate stiffness regulates the neutrophil anti-Candida effector responses and, in turn, how the candida PAMP beta- glucan, affects the generation of traction force. Additionally, tissues are highly confined 3-dimensional spaces and, as such, are different than the 2D tissue culture substrates typically used for in vitro studies. Again, citing prior work from Reichner and colleagues, the physical property of confinement has a significant affect on neutrophil motility and traction. For example, whereas neutrophils are known to require integrins for adhesion and migration on 2D surfaces, integrins become dispensable for migration following entry into the highly confined 3D interstitial space. To study the sensitivity of neutrophils to physical confinement, we developed a double hydrogel compression device and have shown that confinement is the physical trigger for neutrophils to switch to integrin-independent migration and generation of traction. Therefore, although prior work has shown mechanosensitive effects on neutrophil integrin engagement in motility, nothing is known about whether confinement represents a mechanical property within tissues that regulates the anti-Candida effector functions (Aim 2).

项目摘要 /摘要 中性粒细胞是对感染部位的第一批响应者，是抗真菌反应的基础。 这包括口咽念珠菌病的疾病，其中血液中性粒细胞外出成受感染的组织 并发球与入侵念珠菌联系。在此过程中，中性粒细胞不仅对 组织微环境的生化特性，但也具有物理特性。口腔是 来自相对柔软和弹性的舌头，其复合组织的机械性能高度变化 柔软的口感是相对中间刚度的结构，例如食道，以刚性结构 作为硬口味。里希纳实验室和其他实验室表明，人类中性粒细胞对 它们的微环境的机械特征，包括底物刚度。他们报告说人 与软矩阵相比 矩阵。拖曳力的定量和映射显示在僵硬矩阵上产生更大的力 大部分力在细胞后部产生。关于这些发现的一个重大差距是我们没有 有关病原体或其成分弹跳如何影响机械敏感反应的信息 中性粒细胞在整体运动性和在不同刚度的矩阵上产生牵引力的产生。这 很重要，因为是病原体诱导中性粒细胞进入感染组织 作为OPC的原因，抗菌活性和念珠菌感染在口腔中高度相关。这 在此提案中要检验的总体假设是念珠菌的存在会影响机械敏感 人类中性粒细胞的特性。在AIM 1中，我们将使用可调弹性水凝胶来确定底物 刚度调节中性粒细胞抗candida效应子的反应，进而调节念珠菌pAMPβ- 葡萄糖会影响牵引力的产生。 此外，组织是高度限制的3维空间，因此与2D不同 组织培养底物通常用于体外研究。同样，以赖希纳（Reichner）及其同事的先前工作， 监禁的物理特性对中性粒细胞运动和牵引力有重大影响。例如， 众所周知，嗜中性粒细胞需要整合素才能在2D表面上进行粘附和迁移，而整联蛋白成为 进入高度约束的3D间质空间后，可用于迁移。研究的灵敏度 中性粒细胞到物理限制中，我们开发了一种双水凝胶压缩装置，并表明 监禁是中性粒细胞切换到整合素独立迁移和生成的物理触发因素 牵引力。因此，尽管先前的工作已经对中性粒细胞整合素的互动显示了机械敏感的作用 在运动性中，限制是否代表组织中的机械性能，没有什么知道 调节抗Candida效应子功能（AIM 2）。