Understanding neural control of the ocular surface

了解眼表的神经控制

• 批准号: 10586931
• 负责人: MICHAEL W. JENKINS
• 金额: $ 144.46万
• 依托单位: CASE WESTERN RESERVE 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/10586931
• 关键词: 3-Dimensional; Address; Animal Model; Behavior; Brain; Brain region; Calcium; Cells; Chemicals; Cornea; Custom; Data; Debridement; Diabetic mouse; Disease; Dry Eye Syndromes; Dyes; Electrophysiology (science); Environment; Epithelial; Equilibrium; Esthesia; Eye; Eyelid structure; Feedback; Film; Fluorescent in Situ Hybridization; Functional Imaging; Functional disorder; Ganglia; Gland; Goblet Cells; Homeostasis; Human; Image; Image Analysis; Immune; Immune response; Immunology; Implant; Implanted Electrodes; Inflammation; Ion Channel; Keratitis; Knowledge; Lacrimal gland structure; Lead; Light; Measures; Mechanics; Methods; Microscopy; Modeling; Molecular; Morphology; Mus; Natural Immunity; Nerve; Nerve Growth Factors; Nervous system structure; Neurons; Neurosciences; Nociception; Optics; Organ; Organism; Pain; Pain Clinics; Pathologic; Pattern; Peripheral Nervous System Diseases; Production; Pseudomonas aeruginosa; Signal Transduction; Stimulus; Structure; Structure of trigeminal ganglion; Supporting Cell; Surface; System; Techniques; Technology; Thalamic structure; Tissues; Trigeminal System; Work; adaptive immunity; blink reflexes; cell type; chemokine; cytokine; deep learning; density; inflammatory pain; interest; lipid metabolism; lipophilicity; meibomian gland; microscopic imaging; mouse model; multi-electrode arrays; neuroregulation; new technology; ocular surface; painful neuropathy; preservation; receptor; relating to nervous system; response; response to injury; sensory feedback; statistics; tool

Project Summary Currently our understanding of how the nervous system maintains ocular surface homeostasis is extremely limited. New technologies, methods and models are needed to advance our scientific understanding and address knowledge gaps. The ocular surface and tear film-secreting glands (including the lacrimal and meibomian glands, as well as the goblet cells) are carefully controlled to provide an optically smooth, low-scattering surface with appropriate immune and injury responses. Sensory feedback to maintain the structural and functional integrity of the ocular surface is provided by the corneal nerves, which send feedback from stimuli (chemical, thermal, mechanical) to ganglia (e.g., trigeminal) and brain regions (e.g., ventral posteromedial thalamus) to drive production of tear film components as well as the blink reflex. This delicate balance of neural control is disrupted by damage, peripheral neuropathies, inflammation and further complicated by a wide array of immune responses to various diseases. Dysfunction of this feedback loop can lead to a downward spiral of further dysregulation. Aberrant neural control of the ocular surface can lead to abnormal sensation and pain, which in the worst cases can be disabling. To find remedies, it is first essential to understand the underlying neural control system and how it adapts to its environment. In this proposal, we aim to bring new tools and models to study molecular, cellular, and functional interactions across systems responsible for neural control of the ocular surface and examine how they change under different inflammatory and pain conditions. We have assembled an excellent team with expertise across multiple fields including advanced 3D microscopy, neuroscience, electrophysiology, pain, ocular immunology, ocular lipid metabolism, ocular surface disorders, spatial statistics, and machine/deep learning. Here, we will utilize cutting edge techniques and technologies including optical clearing, tract tracing, ethologically-valid behavior analysis, machine/deep learning, spatial statistics, genetically encoded calcium imaging, light-sheet microscopy, multiplexed 3D fluorescence in situ hybridization (FISH) imaging, and multi-array electrodes implanted in the brain. These tools will help us assess molecular, cellular, and functional interactions across organs and begin to understand ocular surface control at the organism level. We will also employ several relevant animal models to assess ocular surface control under different inflammatory and pain conditions. Models include AWAT2 deficient mice that mimic evaporative dry eye disease (DED), diabetic mice, an epithelial debridement model with Pseudomonas aeruginosa that mimics bacterial keratitis, and human donor eyes. The mouse models all have gCaMP6f expressed in corneal nerves allowing functional imaging of calcium transients. With these models we will study both innate and adaptive immunity as well as nociceptive and neuropathic pain responses. In addition, we will apply nerve growth factor (NGF) to our models to study how a potential treatment option alters the ocular surface control system.

项目摘要 目前，我们对神经系统如何维持眼表稳态的理解是极其困难的。 有限公司需要新的技术、方法和模型来推进我们的科学认识和解决 知识差距。眼表面和泪膜分泌腺（包括泪腺和睑板腺， 以及杯状细胞）被小心地控制以提供光学上光滑的、低散射的表面， 适当的免疫和损伤反应。感官反馈，以保持结构和功能的完整性 由角膜神经提供，角膜神经从刺激（化学，热， 机械的）到神经节（例如，三叉神经）和脑区域（例如，腹后内侧丘脑）驱动 泪膜成分的产生以及眨眼反射。这种神经控制的微妙平衡被破坏了 受到损伤、周围神经病变、炎症的影响，并进一步受到广泛的免疫反应的影响， 各种疾病。这种反馈回路的功能失调会导致进一步失调的螺旋式下降。 眼表的异常神经控制可导致异常感觉和疼痛，在最严重的情况下 可能会导致残疾。为了找到补救措施，首先必须了解潜在的神经控制系统， 如何适应环境 在这个建议中，我们的目标是带来新的工具和模型来研究分子，细胞和功能相互作用 跨越负责眼表神经控制的系统，并检查它们在不同的环境下如何变化。 炎症和疼痛状况。我们组建了一支拥有多个领域专业知识的优秀团队 包括先进的3D显微镜、神经科学、电生理学、疼痛、眼免疫学、眼脂质 代谢、眼表疾病、空间统计和机器/深度学习。在这里，我们将利用切割 边缘技术和技术，包括光学清除，道跟踪，行为学有效的行为分析， 机器/深度学习，空间统计，遗传编码钙成像，光片显微镜， 多重3D荧光原位杂交（FISH）成像，以及植入在 个脑袋这些工具将帮助我们评估器官间的分子、细胞和功能相互作用，并开始 了解在生物体水平上的眼表控制。我们还将采用几种相关的动物模型， 评估在不同炎症和疼痛条件下的眼表控制。型号包括AWAT 2缺陷型 模拟蒸发性干眼症（DED）的小鼠、糖尿病小鼠、上皮清创模型， 类似细菌性角膜炎的铜绿假单胞菌和人类供体眼睛。小鼠模型都有 在角膜神经中表达的gCaMP 6 f允许钙瞬变的功能成像。有了这些模型， 将研究先天性和适应性免疫以及伤害性和神经性疼痛反应。此外，本发明还提供了一种方法， 我们将应用神经生长因子（NGF）到我们的模型中，研究潜在的治疗选择如何改变眼部 地面控制系统