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

基本信息

批准号：
10707204
负责人：
Daniel H Kaplan
金额：
$ 150.24万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
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/10707204
关键词：
ATAC-seq Address Affect Afferent Neurons Anatomy Antibodies Architecture Axon Back Binding Proteins Bioinformatics Cell Communication Cells Colon Communication Cornea Corneal Diseases Data Data Set Databases Deposition Dermatology Designer Drugs Development Disease Disease Progression Disease model Dry Eye Syndromes Environment Epigenetic Process Epithelial Cells Epithelium 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 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 Traction Transgenic Mice Vascular Endothelial Growth Factors Viral Wheat Germ Agglutinins Work afferent nerve aqueous cell type chemokine conjunctiva cytokine design designer receptors exclusively activated by designer drugs gain of function immune checkpoint in vivo large datasets loss of function member nerve supply neural neuroimmunology neurotrophic factor novel ocular surface ocular surface disease prevent programs protein expression proteomic signature public database receptor recruit response single-cell RNA sequencing small molecule transcriptomics web portal

项目摘要

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。干眼症和眼表疼痛。了解眼部表面神经如何参与更广泛的眼部健康和疾病。我们概述了一个预计，它将创建大型数据集，以详细说明眼表神经的自然采用系统级别的方法来使用生物信息学和机器学习分析我们的数据平台，以便我们可以甘甘甘甘（Gan Gan Gan Gan Gan Gan）对眼部表面环境健康和疾病。该建议将解决眼部表面神经，上皮细胞和免疫细胞互动。神经免疫学，神经学，蛋白质组学，系统免疫学和生物信息学，我们计划与该团队一起使用。全面分析RFA中概述的所有三个研究级别，其中包括解剖学形态表征，定义神经元和神经元细胞的细胞和分子特性类型并评估神经元和神经元细胞的功能特性。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。在角膜和眼表面支配的神经和非神经元上执行单细胞（SC） - 神经节这些数据将表征神经元身份的身份，并在疾病期间脱离功能。此外，我们将能够分子表征影响您的免疫和上皮细胞疾病期间眼表的功能。健康和疾病期间角膜的蛋白质组学特征。针对神经营养因素的有针对性方法将确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仪分析角膜内神经元和神经元细胞的位置和形态，以便我们可能是能够得出关于神经，上皮和免疫细胞如何在您处紧密相互作用的结论眼表4）我们将使用dreadd（专门由设计师药物激活的设计器受体）扰动神经元敏感性，以便我们可以评估如何改变神经元反应对刺激的反应如何影响眼表稳态和疾病进展。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以后的研究中，所有数据都将存入我们将开发的网站。