Imaging retinal leukocyte-endothelial interaction as a diagnostic marker for multiple sclerosis

项目总结 在多发性硬化症(MS)中，最常见的神经系统疾病是年轻工作的炎性细胞 侵入中枢神经系统(CNS)并攻击对功能至关重要的髓鞘生成细胞 以及轴突的存活。大多数患者都经历了病情不断恶化的复发-缓解病程 神经症状，并最终导致残疾。目前的诊断成像技术无法预测 复发是因为他们无法检测到导致临床疾病的活跃的白细胞渗入中枢神经系统 下游的症状。本提案旨在提供一种方法，使非侵入性评估能够 这种基于检测视网膜中白细胞-内皮相互作用(LEI)的无症状疾病活动 通过实时成像，这将允许预测多发性硬化症复发和快速反馈治疗反应和 疾病的发展。因此，视网膜LEI检测有助于改变MS和 减轻残疾的发展和严重程度。 LEI，一种白细胞渗透所必需的白细胞滚动，在健康的中枢神经系统和 被认为是炎症的标志。我们假设视神经头(ONH)附近的视网膜LEI 在MRI可检测到的病变和临床表现之前，提示活跃的白细胞渗入中枢神经系统 疾病症状。视网膜是中枢神经系统的一个可光学访问的隔室，它将自己借给光学， 通过眼睛的瞳孔进行活体成像。配备扫描激光检眼镜(SLO)，专为 小鼠视网膜成像，在实验性自身免疫性脑脊髓炎的早期检测到视网膜LEI (EAE)，一种公认的多发性硬化症啮齿动物模型，在临床症状出现之前。我们的初步数据进一步证明， 促炎信使可以通过脑脊液(CSF)的运输到达视网膜，并导致 视网膜LEI，尽管视网膜和视神经本身没有发炎。 在目标1中，我们将描述EAE中视网膜LEI的特征，并将其与白细胞渗入EAE的时间进程进行比较。 用流式细胞仪分析CNS，用MRI检测CNS病变的发展。治疗将会 根据LEI检测和疾病时间进程与未经治疗的对照组进行比较。 在目标2中，我们将评估视网膜LEI远程检测CNS炎症的敏感性。我们确定是否 炎症性脑损伤的位置影响视网膜LEI的频率。此外，与MS相关 细胞因子将直接注射到脑脊液中，以测试它们引发视网膜LEI的能力。 在目标3中，我们将开发一种专门用于检测视网膜LEI的自适应光学(AO)SLO。vbl.使用 高速扫描和低音扬声器-高音扬声器AO架构，该仪器将生成 探查视神经头内和附近的区域，预计大多数LEI都在那里。这一尖端技术 该仪器将用于一项初步研究，以评估多发性硬化症患者的视网膜LEI。

PROJECT SUMMARY In multiple sclerosis (MS), the most common neurological disease in the working young, inflammatory cells infiltrate the central nervous system (CNS) and attack the myelin-producing cells that are crucial for function and survival of axons. Most patients experience a relapsing-remitting disease course with ever worsening neurological symptoms and, ultimately, disability. Current diagnostic imaging techniques cannot predict relapses because they cannot detect the active leukocyte infiltration into the CNS that leads to clinical disease symptoms downstream. This proposal intends to provide a method that enables non-invasive assessment of this asymptomatic disease activity based on the detection of leukocyte-endothelial interaction (LEI) in the retina by live imaging, which will allow prediction of MS relapses and rapid feedback on treatment response and disease progression. Thus, retinal LEI detection could help transform therapeutic intervention in MS and mitigate the development and severity of disability in MS. LEI, the rolling of white blood cells that is a requirement for leukocyte infiltration, is absent in healthy CNS and considered a hallmark of inflammation. We hypothesize that retinal LEI near the optic nerve head (ONH) indicates active leukocyte infiltration into the CNS that precedes MRI detectable lesions and clinical disease symptoms. The retina is an optically accessible compartment of the CNS that lends itself to optical, intravital imaging through the pupil of the eye. With a scanning laser ophthalmoscope (SLO), custom built for mouse retinal imaging, retinal LEI was detected in early stages of experimental autoimmune encephalomyelitis (EAE), an accepted rodent model of MS, prior to clinical symptoms. Our preliminary data further demonstrated, that pro-inflammatory messengers can reach the retina via transport of cerebrospinal fluid (CSF) and cause retinal LEI, although the retina and optic nerve themselves are not inflamed. In aim 1, we will characterize retinal LEI in EAE and compare its time course to that of leukocyte infiltration into the CNS, as assessed by flow cytometric analysis, and to development of CNS lesions by MRI. Treatment will be administered upon LEI detection and the disease time course compared with untreated controls. In aim 2, we will assess the sensitivity of retinal LEI to remotely detect CNS inflammation. We determine if the location of an inflammatory brain lesion affects the frequency of retinal LEI. Furthermore, MS-relevant cytokines will be injected directly into the CSF to test their ability to elicit retinal LEI. In aim 3, we will develop an adaptive optics (AO) SLO that is specialized for the detection of retinal LEI. Using high-speed scanning and a woofer-tweeter AO architecture, the instrument will generate a field of view that probes the area in and near the optic nerve head, where most LEI would be expected. This cutting-edge instrument will be used in a pilot study to assess retinal LEI in MS patients.