Assessing how ocular surface nerves, immune cells, and epithelial cells communicate to encourage neuro-immune homeostasis

评估眼表神经、免疫细胞和上皮细胞如何沟通以促进神经免疫稳态

• 批准号: 10595234
• 负责人: Daniel H Kaplan
• 金额: $ 167.85万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 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/10595234
• 关键词: ATAC-seq; Address; Affect; Afferent Neurons; Anatomy; Antibodies; Architecture; Axon; Back; Binding Proteins; Bioinformatics; Cells; Colon; Communication; Cornea; Corneal Diseases; Data; Data Set; Databases; Deposition; Dermatology; Designer Drugs; Development; Disease; Disease model; Dry Eye Syndromes; Environment; Epigenetic Process; Epithelial; Epithelial Cells; Esthesia; Eye; Flow Cytometry; Fostering; Future; Ganglia; Gastrointestinal tract structure; Gene Expression; Genetic; Goals; Graft Rejection; Grant; Hand; Health; Herpesvirus 1; Homeostasis; Human; Imaging Techniques; Immune; Immunohistochemistry; Immunology; Infiltration; Interdisciplinary Study; Intestines; Keratitis; Keratopathy; Knowledge; Label; Lead; Link; Location; Machine Learning; Mass Spectrum Analysis; Mediating; Membrane; Microbiology; Modeling; Molecular; Morphology; Mus; Nature; Nerve; Neuroglia; Neuroimmune; Neurology; Neurons; Neurotransmitters; Ophthalmology; Pain; Pathologic; Pathway interactions; Pattern; Play; Population; Process; Property; Proteins; Proteomics; Records; Regulation; Research; Role; Sensory; Shotguns; Skin; Stimulus; Structure; Structure of superior cervical ganglion; System; Technology; Tissues; Transgenic Mice; Vascular Endothelial Growth Factors; Viral; Wheat Germ Agglutinins; Work; afferent nerve; aqueous; base; cell type; chemokine; conjunctiva; cytokine; design; designer receptors exclusively activated by designer drugs; gain of function; immune checkpoint; in vivo; large datasets; member; nerve supply; neuroimmunology; neuron loss; neurotrophic factor; novel; ocular surface; prevent; programs; protein expression; proteomic signature; public database; receptor; recruit; relating to nervous system; response; single-cell RNA sequencing; small molecule; transcriptomics; web portal

PROJECT SUMMARY The nature and function of ocular surface nerves play a critical role in maintaining ocular surface health while preventing disease. Disruption of ocular surface sensory nerves can lead to blinding keratitis, graft rejection, dry eye disease, and ocular surface pain. The advent of “-omics” studies have provided a platform to understand how ocular surface nerves participate in the broader context of ocular health and disease. Here, we outline a proposal that will create large datasets detailing the nature of ocular surface innervation. We will take a systems-level approach towards analyzing our data using bioinformatics and machine learning platforms, so that we can gain a practical understanding of the ocular surface environment during health and disease. Specifically, this proposal will address how ocular surface nerves, epithelial cells, and immune cells interact. We have recruited a multidisciplinary research team that consists experts in ophthalmology, neuroimmunology, neurology, proteomics, systems immunology, and bioinformatics. With this team, we plan to comprehensively analyze all three levels of research outlined in the RFA, which includes anatomical and morphological characterization, defining cellular and molecular properties of neuronal and non-neuronal cell types, and assessing functional properties of neuronal and non-neuronal cells. Because innervation patterns and functionality changes with disease, we will use multiple ocular surface disease models (viral keratitis and aqueous dry eye disease) to better understand how neurons, epithelial cells, and immune cells affect neuronal functionality and subsequent disease. We will achieve our goal by pursuing four specific aims: 1) We will perform single-cell (sc)-omics on neuonrs and non-neuronal cells of the cornea and ocular surface innervating ganglia. These data will characterize neuronal identities and describe their functionality during disease. Additionally, we will be able to molecularly characterize immune and epithelial cells that influence the functionality of ocular surface afferents during disease. 2) We will use mass spectrometry to analyze the proteomic signatures of the cornea during health and disease. An unbiased shotgun approaches and a targeted approach focusing on neurotrophic factors will be used to identify proteins that influence ocular surface nerve functionality. 3) We will use our imaging techniques and histocytometry to comprehensively analyze the location and morphology of neuronal and non-neuronal cells within the cornea, so that we may be able to make conclusions about how nerves, epithelial, and immune cells interact in close proximity at the ocular surface. 4) We will use DREADD (designer receptor exclusively activated by designer drugs) technology to perturb neuronal sensitivity, so that we can assess how altering neuronal responsiveness to stimuli may affect the ocular surface homeostasis and progress of disease. A systems-level analysis of our observations using machine-learning and bioinformatics will lead to the prediction of novel interaction networks that can be interrogated in future studies. All data will be deposited into a web portal that we will develop.

项目摘要 眼表神经的性质和功能在维持眼表面健康中起着至关重要的作用 预防疾病。眼部表面感觉神经的破坏会导致盲视性角膜炎，移植排斥， 干眼症和眼表疼痛。 “ - 组”研究的冒险为 了解眼部表面神经如何参与更广泛的眼部健康和疾病。这里， 我们将概述一个建议，该提案将创建大型数据集，详细说明眼表面神经的性质。我们将 采用系统级方法使用生物信息学和机器学习分析我们的数据 平台，以便我们可以在健康和 疾病。特别是，该提案将解决眼部表面神经，上皮细胞和免疫细胞如何 相互影响。我们已经招募了一个多学科研究团队，该研究团队是眼科专家， 神经免疫学，神经学，蛋白质组学，系统免疫学和生物信息学。在这个团队中，我们计划 全面分析RFA中概述的所有三个级别的研究，其中包括解剖和 形态学表征，定义神经元和非神经元细胞的细胞和分子特性 类型，并评估神经元和非神经元细胞的功能特性。因为神经支配模式 以及疾病的功能变化，我们将使用多种眼表面疾病模型（病毒性角膜炎和 干眼病），以更好地了解神经元，上皮细胞和免疫细胞如何影响神经元 功能和随后的疾病。我们将通过追求四个具体目标来实现我们的目标：1）我们将 在角膜和眼表面支配的神经细胞和非神经元细胞上执行单细胞（SC） - 神经节。这些数据将表征神经元身份并描述其在疾病中的功能。 另外，我们将能够分子表征影响影响的免疫和上皮细胞 疾病过程中眼表面传入的功能。 2）我们将使用质谱法分析 健康和疾病期间角膜的蛋白质组学特征。公正的shot弹枪接近和 专注于神经营养因素的有针对性方法将用于鉴定影响眼部的蛋白质 表面神经功能。 3）我们将使用我们的成像技术和组织衡量仪全面 分析角膜内神经元和非神经元细胞的位置和形态，以便我们可能是 可以得出关于神经，上皮和免疫细胞如何在近距离近端相互作用的结论 眼表面。 4）我们将使用Dreadd（设计师药物专门激活的设计师接收器）技术 要扰动神经元敏感性，以便我们可以评估如何改变对刺激的神经元反应能力 对我们的观察结果的系统级分析 使用机器学习和生物信息学将导致可以预测的新型交互网络 在未来的研究中受到询问。所有数据都将存入我们将开发的Web门户网站。