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 该提案研究了角膜三叉神经回路，特别强调 角膜上皮与上皮内神经的关系 假设眼表病变涉及短暂和/或长期角膜 上皮屏障破坏会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代偿性降低。单细胞RNA和表观遗传学 将在角膜和三叉神经节细胞中进行分析和原位杂交， 在稳态条件下以及由于干燥导致上皮屏障破坏后投射到角膜 眼睛和角膜上皮清创术。来自这些测定的数据将被整合并单细胞 会生成正常和应激条件下的角膜和三叉神经节图谱 根据独特的表达谱识别细胞。细胞将在这些空间中被解析 组织来识别角膜细胞和神经轴突之间以及三叉神经之间的通讯 神经元和非神经元细胞。在神经节中，时间和空间上解析的转录组 数据将与角膜上皮神经形态和敏感性的变化相关 屏障破坏后发生，以识别维持角膜上皮神经的基因 健康并促进屏障破坏后的病理变化。三个具体目标是 提议研究角膜三叉神经回路。目标 1 将研究屏障的影响 干眼症和上皮清创术对转录组图、角膜上皮的破坏 神经形态和神经功能。目标 2 将研究保存上皮细胞的效果 MMP-9 敲除菌株在干眼和上皮损伤中对这些参数的屏障，以及 目标 3 将研究两种神经敏化趋化因子对正常和正常情况下的神经回路的影响。 眼睛干涩。 所提出的实验研究了角膜上皮-三叉神经感觉回路 协调深层次以确定影响上皮内神经健康和健康的因素 疾病。在这个项目结束时，我们将对机制有更好的理解 角膜上皮细胞和完整屏障抑制神经激活的因素及因素 由屏障破坏的角膜中的角膜细胞产生，导致神经退化和 灵敏度提高。该项目的标杆成果将成为基础 对常见的角膜上皮屏障破坏的影响有了新的认识 角膜疾病对角膜上皮神经网络的完整性和功能的影响。