Mechanical Causation of Corneal Stromal Matrix Synthesis and Fibrosis

角膜基质基质合成和纤维化的机械原因

基本信息

批准号：
10659976
负责人：
Jeffrey W Ruberti
金额：
$ 55.23万
依托单位：
NORTHEASTERN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659976
关键词：
Acceleration Access to Information Address Animals Biopolymers Cell Culture Techniques Cells Cicatrix Collagen Collagen Fibril Communication Connective Tissue Cornea Corneal Stroma Coupled Couples Custom Deposition Development Diffusion Engineering Environment Etiology Event Exclusion Experimental Designs Extracellular Matrix Eye Fibroblasts Fibronectins Fibrosis Genetic Glaucoma Goals Growth Homeostasis Human Image In Vitro Integrins Intervertebral disc structure Investigation Keratoconus Knowledge Laser In Situ Keratomileusis Length Ligaments Measurement Measures Mechanics Medicine Mesenchymal Methods Microscopy Minor Modeling Molecular Morphogenesis Myofibroblast Myopia Natural regeneration Nature Pathologic Pathological Dilatation Pattern Phase Positioning Attribute Production Recording of previous events Retinal Detachment Role Sclera Shapes Speed Stress Structure Tendon structure Testing Tissue Engineering Tissues Traction Tunic Vertebrates Vision Weight-Bearing state Work blebbistatin corneal scar fibrillogenesis improved light transmission live cell microscopy mechanical force mechanical signal member molecular assembly/self assembly monomer organizational structure pressure prospective regenerative response single molecule temporal measurement theories tool wound healing

项目摘要

Project Summary The cornea and sclera are the principal load bearing members in the tough fibrous ocular tunic which we consider to be an integrated mechano-biological structure. During development, the shape of the eye has been tuned to conform to a specific mechanical environment through a long time-scale integration of its loading history with its initial genetic patterning. Although mechanics are known to contribute to the development of many connective tissues, the ocular globe is particularly sensitive to pressure (tensile wall stress) during the expansive phase of growth. Even in the mature ocular tunic, mechanical instabilities often manifest as conditions which disrupt vision (e.g. myopia, keratoconus, post-LASIK ectasia, tractional retinal detachments and glaucoma). While the underlying causes of tissue structural instabilities are poorly understood, we suspect that they are mechanobiological in nature and potentially reflect an imbalance in the tensional homeostasis that exists between mesenchymal cells, their local ECM and the global mechanical environment. We know that during development, disruption of the mechanical connection between fibroblastic cells and their ECM severely retards ocular growth in a manner analogous to pressure loss, suggesting that mechanical communication is critical to proper ocular morphogenesis. However, the effect of mechanical forces on the mechanisms which drive tissue formation and growth are not well characterized. It is remarkable that we still do not fully understand how the most important structural molecule in vertebrates, collagen, is efficiently assembled into highly-organized, functional, load-bearing tissues which are massively expanded into macroscale structures during growth. However, if we are able to uncover new mechanisms which control tissue formation and growth, we will have access to information which can inform therapies for a variety of pathological conditions including fibrosis, myopia, keratoconus and potentially, glaucoma. Additionally, if we understand how tissue is produced, then there will be implications for engineering corneas de novo and for improving approaches to regenerative corneal medicine. In the proposed work, we plan combine our human cell culture model of corneal stromal tissue elaboration with live-cell mechanodynamics imaging to directly observe single collagen molecules during their transition from solution to fibrils. We will thus directly test a new hypothesis which directly couples local and globally applied forces directly to molecular assembly of collagen during fibrillogenesis and growth. The working hypothesis for this proposal is that force causes corneal stromal ECM elaboration to regulate fibril assembly, remodeling and growth. If the hypothesis is correct, there are myriad mechanotherapeutic opportunities and more critically, our basic understanding of collagenous tissue formation and growth, will be fundamentally altered.

项目概要角膜和巩膜是我们认为坚韧的纤维眼外衣中的主要承重构件成为一个综合的机械生物结构。在发育过程中，眼睛的形状已调整为通过长期整合其加载历史来适应特定的机械环境它最初的遗传模式。尽管众所周知，力学对许多领域的发展做出了贡献结缔组织，眼球在扩张期间对压力（张力壁应力）特别敏感成长阶段。即使在成熟的眼外衣中，机械不稳定性也常常表现为以下情况：视力障碍（例如近视、圆锥角膜、LASIK术后扩张、牵拉性视网膜脱离和青光眼）。虽然组织结构不稳定的根本原因尚不清楚，但我们怀疑它们是本质上是机械生物学的，可能反映了存在的张力稳态的不平衡间充质细胞、其局部 ECM 和整体机械环境之间的关系。我们知道，期间成纤维细胞及其 ECM 之间机械连接的破坏严重阻碍了发育眼睛以类似于压力损失的方式生长，表明机械通讯对于适当的眼部形态发生。然而，机械力对驱动组织的机制的影响形成和生长尚不明确。值得注意的是，我们仍然没有完全理解脊椎动物中最重要的结构分子胶原蛋白被有效地组装成高度组织化的、功能性、承重组织，在生长过程中大量扩展为宏观结构。然而，如果我们能够发现控制组织形成和生长的新机制，我们将有获取可以为各种病理状况（包括纤维化）的治疗提供信息的信息，近视、圆锥角膜和潜在的青光眼。此外，如果我们了解组织是如何产生的，那么这将对角膜从头工程和改进角膜再生方法产生影响药品。在拟议的工作中，我们计划结合我们的角膜基质组织的人类细胞培养模型详细阐述活细胞机械动力学成像，直接观察单个胶原分子在其作用过程中的情况从溶液到原纤维的转变。因此，我们将直接测试一个新的假设，该假设直接耦合本地和在原纤维形成和生长过程中，全局直接施加力到胶原蛋白的分子组装。工作中该提议的假设是，力导致角膜基质 ECM 精细化以调节原纤维组装，重塑和成长。如果假设正确，就会有无数的机械治疗机会和更重要的是，我们对胶原组织形成和生长的基本了解将从根本上改变了。