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：
10705272
负责人：
Adam Horn
金额：
$ 10.81万
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/10705272
关键词：
Acute Address Adhesions Architecture Behavior Biophysics Bulla Calcium Signaling Cell Adhesion Cells Chronic Cicatrix Complex Cytoskeleton Dedications Defect Disease Ensure Environment Epidermolysis Bullosa Epithelial Cells Epithelium Equilibrium Extracellular Matrix F-Actin Failure Focal Adhesions Functional disorder Genetic Homeostasis Image Imaging Techniques Injury Link Mechanics Mediating Membrane Minor Modeling Molecular Movement Normal Cell Optics Pathology Patients Phase Phenotype Piezo 1 ion channel Predisposition Process Regulation Regulatory Pathway Research Role Signal Transduction Site Stretching Swelling Syndrome Talin Testing Therapeutic Time Tissues Training Translating Trauma 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 model organism 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.

项目摘要上皮稳态是由作用在整个组织的细胞上的机械力平衡来维持的规模。伤害破坏了这种机械平衡，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。无法有效修复会导致纤维化疤痕，慢性非愈合的伤口，并导致病理学。上皮伤口修复取决于基本的迁移角质形成细胞到损伤部位。虽然众所周知，基本角质形成细胞对机械力敏感，但我们对上皮损伤如何改变体内组织力学以及这些伤口诱导的如何改变组织力学生物物理变化随后协调伤口修复所需的基本角质形成细胞行为。这项研究旨在通过使用幼虫斑马鱼来解决这些问题，斑马鱼适用于实时的浸润成像达到其光学透明度。初步现场模仿实验表明上皮损伤会导致快速碱性角质形成细胞迁移到伤口部位，这是有效修复所需的。基底角质形成细胞迁移是取决于机械信号，例如由于细胞肿胀而引起的膜张力，并且与A有关伤口边缘上皮组织结构的瞬时和局部破坏。基底角质形成细胞迁移可以通过阻止ARP2/3复合激活或通过Talin1敲低来抑制在体内机械信号传导和F-肌动蛋白或局灶性粘合剂复合物重塑之间。此外，瞬时弱化细胞对细胞外基质的粘附改变了基本角质形成细胞的迁移，导致伤口不良康复，并导致上皮结构的长期破坏。这种表型模仿病理学使用Kindler综合征，一种皮肤疾病，患者对损伤的伤口愈合缺陷表现出。这些初步观察表明，幼虫斑马鱼的基本角质形成细胞对机械信号做出反应在受伤后的上皮组织中，通过启动有效增益愈合所需的迁移反应。他们还表明有缺陷的基本角质形成细胞迁移可能导致伤口愈合病理。拟议的研究将调查机械转换器压电和talin1的张力如何调节伤口诱导的F-肌动蛋白和局灶性粘合剂重塑的基本角质形成细胞行为。这些随后将翻译发现以研究Kindler综合征的斑马鱼模型，以确定失调的基本角质形成细胞行为对伤口愈合病理生理学的贡献。确保该项目的成功，已经制定了量身定制的培训计划，以利用出色的研究威斯康星大学 - 麦迪逊大学的环境。斑马鱼模型的使用专门培训用于伤口愈合研究的生物和用于量化上皮组织的体内成像技术力学将有助于完成既定目标。这项培训将促进成功过渡到研究独立性。