Effects of cornea epithelial barrier disruption on the cornea trigeminal neural circuit

摘要 这项建议研究角膜-三叉神经回路，特别强调 角膜上皮与上皮内神经的关系探讨 假设眼表病变涉及一过性和/或延长的角膜 上皮屏障的破坏将导致更大的伤害感受器刺激、不适、疼痛和 上皮神经和间质神经密度代偿性降低。单细胞RNA与表观遗传学 将在角膜和三叉神经节细胞中进行轮廓分析和原位杂交， 在动态平衡条件下和由于干燥造成的上皮屏障破坏后投射到角膜 眼部和角膜上皮清创术。来自这些化验的数据将被整合，并且是单一细胞 将生成正常和应激条件下的角膜和三叉神经节图谱 根据独特的表达模式识别细胞。像元将在这些空间中进行解析 用于识别角膜细胞和神经轴突之间以及三叉神经之间的通讯的组织 神经细胞和非神经细胞。在神经节中，时间和空间分辨的转录 数据将与角膜上皮神经形态和敏感度的变化相关联 发生在屏障破坏后，以识别维持角膜上皮神经的基因 健康促进屏障破坏后的病理变化。三个具体目标是 建议研究角膜-三叉神经环。目标1将调查屏障的影响 干眼和上皮清创术对转录谱、角膜上皮细胞的破坏 神经形态和神经功能。目标2将研究保存上皮细胞的效果 基质金属蛋白酶-9基因敲除株在干眼和上皮损伤中的屏障对这些参数的影响 目的研究两种神经敏化趋化因子对正常和正常大鼠脑内神经回路的影响。 干涩的眼睛。 拟议的实验研究了角膜上皮-三叉神经感觉回路。 协调深层，以确定影响健康和健康的上皮内神经的因素 疾病。在这个项目结束时，我们将对这些机制有一个更好的理解 角膜上皮细胞和完整屏障抑制神经激活的机制及其影响因素 由屏障破坏的角膜中的角膜细胞产生，导致神经退化和 敏感度更高。这个项目的基准成就将是一个基本的 对常见的角膜上皮屏障破坏影响的新认识 角膜疾病对角膜上皮神经网络的完整性和功能的影响。

Abstract This proposal investigates the cornea-trigeminal neural circuit with particular emphasis on the relationship between the cornea epithelium and the intraepithelial nerves to investigate the hypothesis that ocular surface pathologies that involve transient and/or prolonged corneal epithelial barrier disruption will cause greater nociceptor stimulation, discomfort, pain and a compensatory reduction in epithelial and stromal nerve density. Single cell RNA and epigenetic profiling and in situ hybridization will be performed in corneal and trigeminal ganglion cells that project to the cornea in homeostatic conditions and after epithelial barrier disruption due to dry eye and corneal epithelial debridement. Data from these assays will be integrated and single cell atlases of the cornea and trigeminal ganglion under normal and stressed conditions will generated that identify cells based on unique expression profiles. Cells will be spatially resolved in these tissues to identify communications between corneal cells and nerve axons and between trigeminal neuronal and non-neuronal cells. In the ganglion, temporally and spatially resolved transcriptomic data will be associated with the changes in corneal epithelial nerve morphology and sensitivity that occur after barrier disruption to identify genes that maintain the corneal epithelial nerves in health and promote the pathological changes after barrier disruption. Three specific aims are proposed to investigate the cornea-trigeminal circuit. Aim 1 will investigate the effects of barrier disruption from dry eye and epithelial debridement on transcriptomic profiles, corneal epithelial nerve morphology and nerve function. Aim 2 will investigate the effects of preserving epithelial barrier in the MMP-9 knockout strain in dry eye and epithelial wounding on these parameters, and Aim 3 will investigate the effects of two neural sensitizing chemokines on the circuit in normal and dry eyes. The proposed experiments investigate the cornea epithelial-trigeminal sensory circuit at a coordinated deep level to identify factors that impact the intraepithelial nerves in health and disease. At the conclusion of this project, we will have a better understanding of the mechanisms by which corneal epithelial cells and intact barrier suppress nerve activation and the factors produced by cornea cells in barrier disrupted corneas that cause nerve degeneration and heightened sensitivity. The benchmark accomplishments of this project will be a fundamental new understanding of the impact of corneal epithelial barrier disruption that occurs in common corneal diseases on the integrity and function of the corneal epithelial nerve network.