Mechanoregulation of Basal Keratinocyte Migration in Wounded Tissue

受伤组织中基底角质形成细胞迁移的机械调节

基本信息

批准号：
10505700
负责人：
Adam Horn
金额：
$ 10万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10505700
关键词：
Acute Address Adhesions Animal Model Architecture Behavior Biophysics Bulla Calcium Signaling Cell Adhesion Cells Chronic Cicatrix Complex Cytoskeleton Defect Disease Ensure Environment Epidermolysis Bullosa Epithelial Epithelial Cells Equilibrium Extracellular Matrix F-Actin Failure Focal Adhesions Functional disorder Homeostasis Image Imaging Techniques Injury Lead Link Mechanics Mediating Membrane Minor Modeling Molecular Genetics Movement Normal Cell Optics Pathology Patients Phase Phenotype Piezo 1 ion channel Process Regulation Regulatory Pathway Research Role Signal Transduction Site Stretching Swelling Syndrome Testing Therapeutic Time Tissues Training Translating Trauma patient Universities Wisconsin Zebrafish cell behavior chronic wound epithelial injury epithelial wound experimental study fluorescence lifetime imaging healing in vivo in vivo imaging insight intravital imaging keratinocyte knock-down mechanical force mechanical signal mechanotransduction migration non-healing wounds novel prevent repaired response response to injury restoration second harmonic generation imaging skin disorder success tool wound wound closure wound healing

项目摘要

Project Summary Epithelial homeostasis is maintained by the balance of mechanical forces acting upon cells across the tissue- scale. Injury disrupts this mechanical balance and it is unclear how changing homeostatic mechanical signals impacts cell behavior needed for wound repair. Failure to efficiently repair can lead to fibrotic scarring, chronic non-healing wounds, and contribute to pathology. Epithelial wound repair relies on the migration of basal keratinocytes to the site of damage. While it is known that basal keratinocytes are sensitive to mechanical forces, we lack an understanding of how epithelial injury alters tissue mechanics in vivo and how these wound-induced biophysical changes subsequently coordinate basal keratinocyte behavior needed for wound repair. This study aims to address these issues by using larval zebrafish, which are amenable to real-time, intravital imaging due to their optical transparency. Preliminary live-imaging experiments show that epithelial injury causes rapid basal keratinocyte migration to the wound site, which is needed for efficient repair. Basal keratinocyte migration is dependent on mechanical signals, such as membrane tension due to cell swelling, and is associated with a transient and localized disruption of epithelial tissue architecture at the wound edge. Basal keratinocyte migration can be inhibited by blocking Arp2/3 complex activation or through Talin1 knockdown, suggesting a potential link between mechanical signaling and F-actin or focal adhesion complex remodeling in vivo. Further, transiently weakening cell adhesion to the extracellular matrix alters basal keratinocyte migration, causing poor wound healing, and resulting in chronic disruption of epithelial architecture. This phenotype mimics pathology associated with Kindler Syndrome, a skin disease in which patients show wound healing defects in response to injury. These preliminary observations demonstrate that basal keratinocytes of larval zebrafish respond to mechanical signals in epithelial tissue after injury by initiating a migratory response that is required for efficient wound healing. They also suggest that defective basal keratinocyte migration may contribute to wound healing pathology. The proposed study will investigate how tension sensing by the mechanotransducers Piezo1 and Talin1 regulate wound-induced basal keratinocyte behavior by F-actin and focal adhesion remodeling, respectively. These findings will subsequently be translated to investigate a zebrafish model of Kindler Syndrome to determine the contribution of dysregulated basal keratinocyte behavior to wound healing pathophysiology. To ensure the success of this project, a tailored training plan has been developed that takes advantage of the excellent research environment at the University of Wisconsin – Madison. Dedicated training in the use of the zebrafish model organism for wound healing studies and advanced in vivo imaging techniques for quantifying epithelial tissue mechanics will aid in the completion of the stated aims. This training will facilitate a successful transition to research independence.

项目概要上皮稳态是通过作用于整个组织细胞的机械力的平衡来维持的损伤会破坏这种机械平衡，目前尚不清楚如何改变稳态机械信号。影响伤口修复所需的细胞行为，无法有效修复可能导致慢性纤维化疤痕。不愈合的伤口，并有助于上皮伤口修复依赖于基底细胞的迁移。虽然已知基底角质形成细胞对机械力敏感，我们缺乏对上皮损伤如何改变体内组织力学以及这些伤口如何引起的了解生物物理变化随后导致伤口修复所需的基础协调角质形成细胞行为。旨在通过使用斑马鱼幼虫来解决这些问题，由于斑马鱼幼虫易于进行实时活体成像初步的实时成像实验表明，上皮损伤会导致基底细胞快速损伤。角质形成细胞迁移到伤口部位，这是有效修复所需的。依赖于机械信号，例如细胞肿胀导致的膜张力，并且与伤口边缘上皮组织结构的短暂和局部破坏。可以通过阻断 Arp2/3 复合物激活或通过 Talin1 敲低来抑制，这表明潜在的联系机械信号传导与 F-肌动蛋白或粘着斑复合体体内重塑之间的关系进一步是短暂的。细胞与细胞外基质的粘附减弱会改变基底角质形成细胞的迁移，导致伤口不良这种表型模仿了相关的病理学。金德勒综合症是一种皮肤病，患者因受伤而表现出伤口愈合缺陷。初步观察表明斑马鱼幼虫的基底角质形成细胞对机械信号有反应通过启动有效伤口愈合所需的迁移反应，在损伤后的上皮组织中发挥作用。还表明有缺陷的基底角质形成细胞迁移可能有助于伤口愈合病理学。拟议的研究将调查机械传感器 Piezo1 和 Talin1 的张力传感如何调节分别通过 F-肌动蛋白和粘着斑重塑来抑制伤口诱导的基底角质形成细胞行为。研究结果随后将被转化为研究金德勒综合症的斑马鱼模型，以确定基底角质形成细胞行为失调对伤口愈合病理生理学的贡献。该项目取得成功后，我们利用出色的研究成果制定了量身定制的培训计划威斯康星大学麦迪逊分校的环境专门培训斑马鱼模型的使用。用于愈合伤口研究的生物体和用于量化上皮组织的先进体内成像技术机械师将有助于完成既定目标，该培训将有助于成功过渡到。独立研究。