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

描述(申请人提供)：老年性黄斑变性(AMD)是60岁以上人群失明的主要原因。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)的疗效因患者而异，持续的反应是不可预测的。迫切需要开发新的功能生物标志物来a)预测哪些患者将对抗血管内皮生长因子治疗有反应，b)确定谁将需要再次治疗以及他们应该多久治疗一次，以及c)确定治疗难治的湿性黄斑变性的候选治疗靶点。我们提出了一个新的假设，即在湿性黄斑变性中，活化的VEGFR受体的磷酸化形式以及与新生血管、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进入视网膜的玻璃体重塑。我们的初步研究有力地证实了这一假设，并为这个为期2年的探索性开发项目的目标提供了理论基础。目的1：验证老年性湿性黄斑变性患者的玻璃体中存在磷酸化形式的蛋白受体(如VEGFR)和其他信号通路蛋白的假说。我们将比较50个有效的磷酸化受体蛋白和信号通路分析物参与新生血管形成(例如VEGFR、PDGFR)的水平，以及与血管通透性、缺氧、炎症、存活/凋亡和疾病中的粘连相关的下游通路在治疗前后的水平。目的：探讨湿性黄斑变性患者玻璃体中AIM-1蛋白标志物与临床疗效的关系。目的3：完善的新型磷化蛋白亲和捕获技术作为一种通用的方法来发现玻璃体中这类新的磷酸化蛋白的额外成员并进行MS测序，这些成员可能构成下一代功能生物标记物。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，最大限度地扩大未来将研究结果转化为常规临床应用的机会。B)范式转换假说，预测玻璃体中存在一类新的功能相关的生物标志物；c)独特的临床研究研究集，包括在3至6个周期的抗VEGF眼内治疗前后收集的50名患者的玻璃体样本；d)根据CAP/CLIA指南测量高灵敏度和高精度的新技术；以及e)亲和捕获和浓缩玻璃体中的磷蛋白的新技术，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记物的临床可行性和治疗相关性。年龄相关性黄斑变性(AMD)是60岁以上人群失明的主要原因。这项提议将开发新的玻璃体生物标记物，以a)预测哪些患者将对抗血管内皮生长因子治疗有反应，b)确定谁将需要再次治疗以及他们应该多久治疗一次，以及c)确定治疗难治的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.