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

角膜上皮屏障破坏对角膜三叉神经回路的影响

• 批准号: 10708182
• 负责人: RUI CHEN
• 金额: $ 117.83万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10708182
• 关键词: Apical; Atlases; Axon; Basement membrane; Benchmarking; Biological Assay; Blinking; Brain Stem; CXCL1 gene; CXCL10 gene; Cell membrane; Cells; Chemicals; Chromatin; Clinical; Communication; Complement; Complex; Cornea; Corneal Abrasion; Corneal Diseases; Cues; Data; Debridement; Desiccation; Disease; Epithelial Cells; Epithelium; Esthesia; Event; Eye; Film; Ganglia; Gelatinase B; Gene Activation; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic Transcription; Goals; Health; Homeostasis; Immune; In Situ Hybridization; Injury; Knock-out; Measures; Mechanics; Molecular; Morphology; Nerve; Nerve Degeneration; Nerve Pain; Neuroglia; Neurons; Neuropeptides; Nociceptors; Optic Nerve; P2X-receptor; Pain; Pathologic; Pathology; Peptide Hydrolases; Peptides; Peripheral; Process; Production; Proliferating; Protein Family; RNA; Radial; Schwann Cells; Sclera; Sensory; Signal Transduction; Sodium Channel; Stimulus; Stress; Structure of trigeminal ganglion; Surface; Synapses; Thinness; Tight Junctions; Time; Tissues; Trauma; Travel; Trigeminal System; Trigeminal nerve structure; Vanilloid; chemokine; corneal epithelium; cytokine; density; environmental stressor; epigenetic profiling; epithelial injury; epithelial wound; experimental study; eye dryness; ganglion cell; intraepithelial; irritation; microbial; nerve supply; neural; neural circuit; neuronal cell body; neurosensory; ocular pain; ocular surface; preservation; receptor; recruit; response; transcriptomics; voltage; wound healing

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.

抽象的 该提案调查了角膜 - 三元神经回路，特别强调 角膜上皮和上皮内神经之间的关系 假设涉及短暂和/或长角膜的眼表面病理 上皮屏障破坏将导致更大的伤害感受器刺激，不适，疼痛和 上皮和基质神经密度的补偿性降低。单细胞RNA和表观遗传学 分析和原位杂交将在角膜和三叉神经节细胞中进行 在稳态条件下以及由于干燥而导致上皮屏障破坏后向角膜进行项目 眼睛和角膜上皮清创术。这些分析的数据将集成并单元格 在正常和压力条件下，角膜和三叉神经节的地图集将产生 根据唯一的表达曲线识别细胞。细胞将在这些中在空间上解决 组织以识别角膜细胞与神经轴突之间以及三叉神经之间的通信 神经元和非神经元细胞。在神经节中，时间和空间分辨的转录组 数据将与角膜上皮神经形态和灵敏度的变化有关 在障碍破坏后，发生这种情况，以鉴定维持角膜上皮神经的基因 健康并促进障碍破坏后的病理变化。三个具体目标是 提议研究角膜 - 三元电路。 AIM 1将研究障碍的影响 眼睛干燥和上皮清创术的破坏，角膜上皮 神经形态和神经功能。 AIM 2将研究保留上皮的影响 在这些参数上，在干眼症和上皮伤口中的MMP-9敲除应变中的障碍物， AIM 3将研究两种神经敏化趋化因子对正常电路的影响，并且 干眼睛。 提出的实验研究了在A处的角膜上皮 - 三级感觉电路 协调深层的水平，以确定影响健康上皮内神经的因素 疾病。在该项目的结论中，我们将对机制有更好的了解 角膜上皮细胞和完整的屏障抑制神经激活和因素 由屏障中的角膜细胞产生的破坏角膜，导致神经变性和 提高灵敏度。该项目的基准成就将是一个基本 对角膜上皮屏障破坏的影响的新了解 角膜疾病关于角膜上皮神经网络的完整性和功能。