Phosphorylated signal pathway proteins: A new class of vitreous biomarkers for op

描述（由申请人提供）：视网膜相关性黄斑变性（AMD）是60岁以上个体失明的主要原因。新的血管内皮生长因子（VEGF）抑制剂治疗年龄相关性湿性黄斑变性（AMD）的疗效在不同患者之间差异很大，并且持续的反应是不可预测的。非常需要开发新的功能性生物标志物，以a）预测哪些患者对抗VEGF治疗有反应B）确定哪些患者需要再治疗以及他们应该多久治疗一次，以及c）鉴定治疗难治性湿性黄斑变性中的候选治疗靶标。我们提出了一个新的假说，即激活的VEGFR受体的磷酸化形式和与新生血管形成、血管通透性、细胞凋亡和炎症相关的下游或相互连接的信号蛋白，将在湿性黄斑变性中脱落到视网膜的玻璃体重塑中。我们的初步研究有力地证实了这一假设，并为这一为期2年的探索性开发项目的目标提供了依据。目的1：检验磷酸化形式的蛋白受体（如VEGFR）和其他信号通路蛋白脱落到年龄相关性湿性黄斑变性患者的玻璃体中的假设。我们将在治疗前后比较疾病与对照组中参与新生血管形成（例如VEGFR、PDGFR）的50种经验证的磷酸化受体蛋白和信号通路分析物以及与血管通透性、缺氧、炎症、促存活/凋亡和粘附相关的下游通路的水平。目标二：评估湿性黄斑变性患者玻璃体中鉴定的Aim 1蛋白标志物是否与治疗的临床反应相关。目标3：完美的新型磷酸化蛋白亲和捕获技术作为一种通用方法来发现和MS测序玻璃体中磷酸化蛋白的这种新型生物标志物类别的其他成员，这些成员可能构成下一代功能性生物标志物。创新的组成部分是a）乔治梅森大学和国家视网膜研究所之间的协同合作，最大限度地提高了未来将研究结果转化为常规临床应用的机会。B）预测玻璃体中存在一类新的功能相关生物标志物的范式转变假说，c）独特的临床研究集，包括在3至6个周期的抗VEGF眼内治疗之前和之后从50名患者收集的玻璃体样品，d）根据CAP/CLIA指南测量20微升中的新型磷酸蛋白分析物组的新型高灵敏度和精确技术，和e）用于玻璃体中磷蛋白的亲和捕获和富集的新技术，作为未来的发现工具。总体目标是证明一类新的玻璃体生物标志物的临床可行性和治疗相关性。视网膜相关性黄斑变性（AMD）是60岁以上个体失明的主要原因。该提案将开发新的玻璃体生物标志物，以a）预测哪些患者对抗VEGF治疗有反应B）确定哪些患者将需要再治疗以及他们应该多久治疗一次，以及c）鉴定治疗难治性湿性黄斑变性的候选治疗靶点。

DESCRIPTION (provided by applicant): Age-related macular degeneration (AMD) is the leading cause of blindness in individuals over age 60. The efficacy of the new vascular endothelial growth factor (VEGF) inhibitors for treatment of age- related wet macular degeneration (AMD) is highly variable from patient to patient, and sustained responses are unpredictable. There is a great need to develop novel functional biomarkers to a) predict which patients will respond to anti-VEGF therapy b) determine who will need re-treatment and how often they should be treated, and c) identify candidate therapeutic targets in wet macular degeneration that is refractory to therapy. We propose the new hypothesis that phosphorylated forms of the activated VEGFR receptors and downstream or interconnected signaling proteins associated with neovascularization, vascular permeability, apoptosis, and inflammation, will be shed into the vitreous remodeling of the retina in wet macular degeneration. Our preliminary studies have strongly confirmed this hypothesis and provide the rationale for the Aims of this 2 year exploratory development project. Aim 1:Test the hypothesis that phosphorylated forms of protein receptors (e.g. VEGFR) and other signal pathway proteins are shed into the vitreous of patients with age-related wet macular degeneration. We will compare the levels of 50 validated phosphorylated receptor proteins and signal pathway analytes involved in neovascularization (e.g.VEGFR, PDGFR), and downstream pathways associated with vascular permeability, hypoxia, inflammation, prosurvival/apoptosis, and adhesion in disease versus control, before and after treatment. Aim 2: Evaluate if the protein markers of Aim 1 identified in the vitreous of wet macular degeneration patients, correlate with clinical response to therapy. Aim 3: Perfect novel phospho-protein affinity capture technology as a general method to discover and MS sequence additional members of this novel biomarker class of phosphorylated proteins in the vitreous that may constitute the next generation of functional biomarkers. The innovative components are a) a synergistic collaboration between George Mason University and the National Retinal Institute that maximizes opportunities for future translation of the findings to routine clinical use. b) A paradigm-shifting hypothesis that predicts the existence of a novel class of functionally relevant biomarkers in the vitreous, c) A unique clinical research study set comprising vitreous samples collected from 50 patients before and after 3 to 6 cycles of anti-VEGF intraocular therapy, d) Novel highly sensitive and precise technology to measure under CAP/CLIA guidelines the novel phospho- protein analyte panel in 20 microliters, and e) novel technology for affinity capture and enrichment of phosphoproteins in vitreous as a discovery tool for the future. The overall objective is to demonstrate the clinical feasibility and treatment associated relevance for a new class of vitreous biomarkers.Age-related macular degeneration (AMD) is the leading cause of blindness in individuals over age 60. This proposal will develop novel vitreous biomarkers to a) predict which patients will respond to anti-VEGF therapy b) determine who will need re-treatment and how often they should be treated, and c) identify candidate therapeutic targets in wet macular degeneration that is refractory to therapy.