Blink, Lacrimation, and Nociception: Precision Mapping and Integrated Atlas Generation of Corneal Trigeminal Afferents

眨眼、流泪和伤害感受：角膜三叉神经传入的精确绘图和集成图谱生成

• 批准号: 10707309
• 负责人: Sina Farsiu
• 金额: $ 130.57万
• 依托单位: DUKE UNIVERSITY
• 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/10707309
• 关键词: 3-Dimensional; Ablation; Anatomy; Atlases; Axon; Behavioral; Biological Assay; Blinking; Brain Stem; Catalogs; Categories; Cells; Classification; Clinical Research; Color; Complex; Computer software; Cornea; Coupled; Data; Data Set; Development; Dimensions; Dissection; Dry Eye Syndromes; Electrophysiology (science); Eye; Foundations; Future; Generations; Genes; Genetic; Genomics; Grant; Homeostasis; Human; Image; Image Analysis; Immunofluorescence Immunologic; Infection; Ions; Knowledge; Label; Lacrimation; Link; Machine Learning; Macrophage; Maintenance; Maps; Mediating; Microscopy; Modeling; Molecular; Morphology; Mouse Strains; Mus; Nature; Nerve; Nerve Fibers; Nerve Plexus; Neurons; Neuropathy; Nociception; Nociceptors; Output; Pain; Pattern; Population; Process; Production; Property; Pseudomonas aeruginosa; Quality of life; Reflex action; Resolution; Signal Transduction; Structure; Structure of trigeminal ganglion; Tissues; Translational Research; Trigeminal System; Validation; Work; analog; artificial intelligence method; automated segmentation; behavior prediction; blink reflexes; density; differential expression; experimental study; eye dryness; genetic approach; grasp; human subject; imaging Segmentation; machine learning classification; multidisciplinary; multiple datasets; neuronal circuitry; neurotransmission; new therapeutic target; novel strategies; ocular pain; ocular surface; open source; patch clamp; pharmacologic; reflectance confocal microscopy; response; segmentation algorithm; transcriptome; transcriptome sequencing

ABSTRACT Corneal nerves that mediate pain, blink reflexes and tear production are indispensable in the proper maintenance of ocular surface homeostasis. However, the complexity of these neurons, both at the axon level in the cornea, and cell bodies in the trigeminal ganglion, has made it increasingly difficult to grasp their full nature, resulting in key knowledge gaps in the field. Consistent with the call for the current U01, we will address this barrier via comprehensive analyses of corneal nerves at the morphologic, molecular, and functional level. We have assembled a multidisciplinary team with complementary expertise, enabling integrated analyses of spatial, electrophysiologic, genomic and behavioral profiles in mice; and AI-assisted structure-function studies in human subjects, to open new inroads in the field. In Aim 1, we will anatomically and functionally map corneal-projecting trigeminal afferents in mice. Genetic approaches will be applied to rigorously profile the spatial arrangement of nociceptors in the cornea and trigeminal ganglion using seven- color immunolabeling. This spatial information will be directly linked with the electrophysiological profiling of these respective populations. In Aim 2, genomic analyses of corneal-projecting trigeminal afferents will be conducted applying a new platform for spatial RNA-seq at cellular resolution. We will use mouse strains to parse out the transcriptomes and behavioral outputs at the genetic level. Moreover, we apply two novel approaches to selectively target corneal nerves involved in tear production versus blinking. In Aim 3, we will discover morphological patterns of corneal nerves that predict blinking, lacrimation, and nociception in humans. We will image corneal nerves with in vivo confocal microscopy of subjects with differential blink, tear, and nociceptive behavioral outputs, thereby capturing functional analogs of the mouse experiments in Aims 1 and 2. With the AI-based auto-segmentation algorithm that we are developing, we will be able to apply machine learning for multidimensional profiling of nerve patterns, and then compare these with respective behavioral outputs. These AI-guided efforts will provide critical clues for understanding corneal afferent structure-function in humans. In summary, our collective studies will lead to an unprecedented cartography of corneal afferents in blink, lacrimation, and nociception. The advancements from this work will be poised to facilitate a deeper understanding of related pathobiology including neuropathic ocular pain and dry eye disease that will lay the foundation for future translational and clinical research. All genomic, imaging and electrophysiologic datasets produced will be made publicly available, and all software products for corneal nerve image segmentation will be made freely available online as open-source and easy-to-use software packages.

抽象的 介导疼痛，眨眼反射和泪水产生的角膜神经在适当的 维持眼表稳态。但是，这些神经元的复杂性，都在轴突水平 在角膜和三叉神经节中的细胞体中，越来越难以掌握它们 自然，导致该领域的关键知识差距。与当前U01的电话一致，我们将 通过在形态学，分子和 功能水平。我们已经组建了一个具有互补专业知识的多学科团队 小鼠的空间，电生理，基因组和行为谱的综合分析；并辅助 在人类受试者中进行结构 - 功能研究，以在该领域开放新的侵害。在AIM 1中，我们将在解剖学上 并在功能上绘制小鼠的角膜预测三叉神经传入。遗传方法将应用于 严格介绍了使用七个 - 颜色免疫标记。此空间信息将直接与 这些各自的人群。在AIM 2中，角膜预测三叉神经传统的基因组分析将是 在细胞分辨率下应用新的空间RNA-seq的平台进行了操作。我们将使用鼠标菌株 在遗传水平上解析转录组和行为输出。而且，我们应用了两本小说 选择性地靶向参与泪水产生与眨眼的角膜神经的方法。在AIM 3中，我们将 发现角膜神经的形态学模式，可预测人类闪烁，泪和伤害感。 我们将对具有不同眨眼，撕裂和 伤害性行为输出，从而在AIM 1和 2。使用我们正在开发的基于AI的自动分割算法，我们将能够应用机器 学习神经模式的多维分析，然后将它们与各自的行为进行比较 输出。这些AI引导的努力将为了解角膜传入结构功能提供关键线索 在人类中。总而言之，我们的集体研究将导致角膜传入的前所未有的制图 眨眼，泪和伤害感受。这项工作的进步将有望促进更深入 了解相关病理生物学，包括神经性眼部疼痛和干眼症，这将放置 未来翻译和临床研究的基础。所有基因组，成像和电生理数据集 生产将公开可用，所有用于角膜神经图像分割的软件产品将 可在线免费提供，作为开源和易于使用的软件包。