In Vivo Function and Metabolism Evaluation of Glaucomatous RGCs by Two-Photon Scanning Laser Ophthalmology

双光子扫描激光眼科评价青光眼 RGC 的体内功能和代谢

基本信息

批准号：
10660761
负责人：
Yang Hu
金额：
$ 64.15万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2028-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660761
关键词：
Acute Anatomy Angle-Closure Glaucoma Animal Model Animals Belief Biological Markers Biosensor Blindness Blood Vessels Cell physiology Cells Cessation of life Characteristics Chronic Color Combined Modality Therapy Contact Lenses Cornea Data Dehydration Detection Development Disease Disease Progression Electrophysiology (science)Evaluation Excision Fluorescence Functional Imaging Functional disorder Fundus photography Glaucoma Image Imaging Techniques Individual Interdisciplinary Study Investigation Label Lasers Light Lighting Metabolic Metabolism Modeling Monitor Morphology Motion Mus Nerve Degeneration Neural Retina Neurons Ocular Hypertension Open-Angle Glaucoma Ophthalmology Ophthalmoscopy Optics Pattern Phenotype Photic Stimulation Photons Photoreceptors Physiologic Intraocular Pressure Physiology Predisposition Property Protocols documentation Retina Retinal Diseases Retinal Ganglion Cells Scanning Severities Silicone Oils Spottings System Testing Time Transgenic Mice Traumatic injury Update Visualization calcium indicator cell type cellular imaging clinically relevant experimental study imaging modality imaging platform imaging system improved in vivo in vivo imaging in vivo optical imaging neural circuit neuroprotection non-invasive imaging novel overexpression prevent resilience response retinal imaging retinal stimulation sensor two-photon visual processing visual stimulus

项目摘要

PROJECT SUMMARY Glaucoma, the most common worldwide cause of irreversible blindness, is characterized by progressive dysfunction and death of retinal ganglion cells (RGCs). We recently employed confocal scanning laser ophthalmoscopy (cSLO) to successfully obtain in vivo Ca2+ imaging with mouse RGCs expressing jGCaMP7s, a genetically encoded calcium indicator. Thousands of ON, OFF, and ON-OFF RGCs with characteristic responses to light stimulation are readily detected in living animals through this non-invasive in vivo imaging platform. Here we seek to develop a more advanced, first-of-its-kind two-photon (2P)-SLO platform with patterned stimulation and multiple detection channels. Through a multidisciplinary collaboration with expertise in in vivo optical imaging, RGC pathophysiology, and retinal neural circuitry and visual processing, we will use complementary imaging techniques and state-of-the-art analysis protocols to understand naïve RGC physiology in real time. We recently extended our original mouse silicone oil-induced pupillary blocking and ocular hypertension (SOHU) model to recapitulate phenotypes of two forms of glaucoma: a chronic model with moderate IOP elevation and mild RGC neurodegeneration; and an acute model with greatly elevated IOP and severe neurodegeneration. Importantly, SO removal reduces IOP to normal almost immediately, allowing better exploration of the effects of IOP lowering treatment and combined treatment with neuroprotection strategies. Thus, we will determine the longitudinal functional and metabolic changes of glaucomatous RGCs, under clinically relevant models, both before and after IOP normalization and/or neuroprotective treatments. These data will deepen our understanding of the pathophysiology of glaucoma, towards finding much-sought biomarkers to better predict progression, and create more relevant endpoints for developing treatment to restore RGC physiology in vivo.

项目摘要青光眼是全球最常见的不可逆失明的原因，其特征是进步残留神经节细胞（RGC）的功能障碍和死亡。我们最近进行了共聚焦扫描激光眼镜检查（CSLO），用表达JGCAMP7S的小鼠RGC成功获得体内Ca2+成像，A 遗传编码的钙指标。具有特征性响应的数千个ON，OFF和ON-OFF RGC 通过这个非侵入性的体内成像平台，在活的动物中很容易检测到光刺激。这里我们试图开发一个具有图案刺激的更先进的，最先进的两光子（2p）-slo平台和多个检测通道。通过与体内光学专业知识的多学科合作成像，RGC病理生理学以及视网膜神经回路和视觉处理，我们将使用完整性成像技术和最先进的分析方案，以实时了解RGC生理。我们最近扩展了我们原始的小鼠硅油诱导的瞳孔阻塞和眼高血压（SOHU）概括两种形式的青光眼的表型的模型：一种具有中等IOP升高和的慢性模型轻度RGC神经变性；以及具有较高IOP升高和严重神经变性的急性模型。重要的是，因此几乎立即将IOP降低到正常，从而更好地探索了 IOP降低治疗并将治疗与神经保护策略结合在一起。那我们将确定青光眼RGC的纵向功能和代谢变化，在临床相关模型下，都 IOP归一化和/或神经保护治疗前后。这些数据将加深我们的理解青光眼的病理生理学，寻找备受瞩目的生物标志物以更好地预测进展，并且创建更相关的终点，以开发在体内恢复RGC生理的治疗方法。