Complement in AMD: Mechanisms and Therapeutic Intervention

AMD 中的补体：机制和治疗干预

基本信息

批准号：
8039646
负责人：
JOHN D LAMBRIS
金额：
$ 63.34万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8039646
关键词：
Address Age related macular degeneration Alternative Complement Pathway Animal Model Binding Biochemical Biological Assay Birds Blindness C3 gene Chemicals Complement Complement Activation Complement Component Gene Complement Factor B Complement Factor H Complement Inactivators Complex Computer Simulation Computing Methodologies Development Disease Disease Progression Disease model Drug Delivery Systems Drug Kinetics Drusen Elderly Etiology European Eye Gel Genes Genetic Genetic Polymorphism Genotype Goals High Prevalence Human Individual Inflammatory Knowledge Laboratories Length Link Macaca fascicularis Macular degeneration Mammalian Cell Mass Spectrum Analysis Methods Modeling Modification Molecular Monkeys Mutagenesis Mutation Pathogenesis Pathway interactions Patients Peptides Pharmaceutical Preparations Plasma Post-Translational Protein Processing Primates Process Property Proteins Regulation Research Retinal macula Risk Risk Factors Role Sampling Series Severity of illness Solutions Staging Structure Testing Therapeutic Therapeutic Intervention Translating United States Variant base clinically relevant complement system compstatin effective therapy improved inhibitor/antagonist mutant prevent weapons

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to elucidate the role of complement in the pathogenesis of age-related macular degeneration (AMD), and to identify complement inhibitors that prevent disease progression. Though AMD has emerged as the predominant cause of blindness in elderly people, our knowledge of the underlying molecular processes and the availability of treatment options remain largely limited. Accumulating evidence suggests a crucial role for complement in the progression of AMD, and several genotyping studies have identified polymorphisms in the genes for C3, factor B, and factor H as important risk factors for development of this disease. However, the functional significance of these genetic findings remains unknown and need to be translated into a disease model. To elucidate the functions of complement in AMD on the protein level, we propose two specific aims. In Aim 1 we perform a comprehensive in-depth analysis of complement proteins and their AMD- associated alloforms, by expressing them in mammalian cells and by isolating them from the plasma of AMD patients and healthy individuals. Sequence modifications will be determined by mass spectrometry, and the direct binding and functional activities of all individual proteins and their various combinations will be systematically tested in a panel of well-established biophysical and biochemical assays. Finally, the effect of these protein modifications on the structure and contact interface of the involved complement components will be analyzed by correlating our findings with available crystal structures, solution-based structural analysis, and computational models. The development and testing of AMD-targeting drugs is often hampered by restricted access to disease-relevant animal models and unfavorable pharmacokinetic profiles. In Aim 2, we will utilize a clinically relevant monkey model of macular degeneration for testing the effect of complement inhibitors on disease progression. The peptidic inhibitor compstatin will be injected intravitreally and its effect on drusen formation will be evaluated. Furthermore, a sustained drug release in the eye as a result of intravitreal gel formation will be explored. The systematic assessment of the functional consequences for AMD-associated complement modifications and the testing of complement inhibition in a disease-relevant animal model will contribute to our understanding of AMD, as well as contribute to the development of more effective treatments. PUBLIC HEALTH RELEVANCE: The purpose of our study is to describe the contributions of complement activation and inhibition to the pathogenesis of age-related macular degeneration. This will be accomplished by systematically evaluating the effect of disease-related polymorphisms on the structure, binding, and function of complement components, and by assessing the impact of complement inhibitors on disease progression in a monkey model of macular degeneration. Thus, these studies will likely improve our capacity for describing and treating the disease.

描述（由申请人提供）：该项目的长期目标是阐明补体在与年龄相关的黄斑变性（AMD）发病机理中的作用，并确定可以预防疾病进展的补体抑制剂。尽管AMD已成为老年人失明的主要原因，但我们对基本分子过程的了解和治疗方案的可用性仍然在很大程度上有限。积累的证据表明，补体在AMD的进展中起着至关重要的作用，并且一些基因分型研究确定了C3，因子B和因子H的基因中的多态性，这是该疾病发展的重要风险因素。但是，这些遗传发现的功能意义仍然未知，需要转化为疾病模型。为了阐明AMD在蛋白质水平上的补体功能，我们提出了两个具体目标。在AIM 1中，我们通过在哺乳动物细胞中表达它们，并通过将它们与AMD患者和健康个体的血浆中分离出来，对补体蛋白及其相关的同种异体进行全面的深入分析。序列修饰将由质谱法确定，并且所有单个蛋白质及其各种组合的直接结合和功能活性将在成熟的生物物理和生化测定面板中系统地测试。最后，通过将我们的发现与可用的晶体结构，基于溶液的结构分析和计算模型相关联，将分析这些蛋白质修饰对相关补体组件结构和接触界面的影响。易于疾病的动物模型和不利的药代动力学特征，通常会阻碍靶向AMD靶向药物的开发和测试。在AIM 2中，我们将利用临床上相关的黄斑变性猴模型来测试补体抑制剂对疾病进展的影响。肽抑制剂Compstatin将被注射玻璃体内，并将评估其对Drusen形成的影响。此外，将探索玻璃体内凝胶形成的持续药物释放。对与AMD相关的补体修饰的功能后果的系统评估以及与疾病相关的动物模型中补体抑制的测试将有助于我们对AMD的理解，并有助于发展更有效的治疗方法。公共卫生相关性：我们研究的目的是描述补体激活和抑制与年龄相关的黄斑变性的贡献。这将通过系统地评估与疾病相关的多态性对补体成分的结构，结合和功能的影响，以及评估补体抑制剂对猴子变性模型中疾病进展的影响。因此，这些研究可能会提高我们描述和治疗疾病的能力。