Strategies to Control Alternative Pathway Activation in Age-Related Sensory Loss.

控制与年龄相关的感觉丧失中替代途径激活的策略。

基本信息

批准号：
10652274
负责人：
Nathaniel Bryan David Parsons
金额：
$ 4.74万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10652274
关键词：
Abate Abnormal Macrophage Age Age related macular degeneration Alternative Complement Pathway Anaphylatoxins Animal Model Animals Annexin A4 Antibodies Auditory system Binding Biodistribution Biological Biological Assay Blindness Cell surface Cells Chimeric Proteins Clinical Cochlea Competence Complement Complement 3 Convertase Complement 3d Receptors Complement Activation Complement Factor H Complement Membrane Attack Complex Complex Deposition Diet Disease Environment Epitopes Etiology Expression Profiling Eye Feedback Gene Activation Goals Histologic Histology Human Immune Immunohistochemistry Inflammation Inflammatory Lead Learning Macrophage Activation Mediating Membrane Modeling Molecular Mus Nerve Degeneration Nerve Tissue Neurodegenerative Disorders Neurosciences Research Ocular Pathology Ophthalmology Opsonin Pathologic Pathology Pathway interactions Pattern Persons Phase Postdoctoral Fellow Presbycusis Proteins RNA Reporting Research Research Training Schools Sensory Site Smoke Smoking Stress Structure of retinal pigment epithelium System Techniques Technology Testing Tissue Preservation Tissues Training Translational Research Vision Visual system structure Work adeno-associated viral vector age related aged aging auditory system base cytokine extracellular gene therapy graduate student human model human tissue inhibitor interest macrophage man mouse model natural antibodies neoantigens nerve damage novel post-doctoral training prevent promoter response to injury sensory neuroscience tissue repair treatment strategy vector vector control

项目摘要

PROJECT SUMMARY/ABSTRACT Dual sensory loss is defined as a combination of age-related vision loss, e.g., age-related macular degenera- tion (AMD) and age-related hearing loss (ARHL) that co-occurs in people age >65. AMD, dry and wet forms, and ARHL are multifactorial diseases that share etiologies such as smoking and complement dysregulation. Importantly, the amplification loop of the complement alternative pathway (AP) is crucial to control, as it is re- sponsible for the majority of complement activation on cell surfaces and extracellular membranes. The AP is inhibited by circulating complement protein factor H (fH). One of the mechanisms by which complement is acti- vated is natural antibodies (nAbs) binding to neoepitopes, or damage-associated molecular patterns, on dam- aged tissues in response to injury. The complement effector molecules and nAbs activate macrophages and other immune cells, intensifying the inflammatory state that can lead to neurodegeneration. Two well-studied fusion proteins, CR2-fH and B4-scFv-fH, will be used to target the inhibitory domain of fH to sites of tissue damage. The complement receptor 2 (CR2) domain binds to complement fragment deposited sites, and the single chain antibody B4 (B4-scFv) domain binds to modified annexin IV exposed on damaged tissues. Both CR2-fH and B4-scFv-fH have been shown to be efficacious in mouse models of neurodegenerative diseases when administered systemically, or for CR2-fH locally and via gene therapy. Of interest is that complement and macrophage activation are elevated in the subretinal space of wet AMD mouse models and in cochlear tissue of aged mice. In addition, smoke-induced proinflammatory, neurodegenerative AP activation, and abnormal macrophage activity, have been seen the eye and cochlea. Based on these observations, we hypothesize that vector driven CR2-fH and injected B4-scFv-fH localize fH to damaged nerve tissues and mitigate wet AMD and ARHL pathology, in part by reducing macrophage activation, and thereby reducing the complement- macrophage inflammatory feedback loop. I will test this hypothesis in two specific aims, the results of which will transition to my postdoctoral research. Aim 1 will provide proof of concept that the C3 promoter is modulated in a complement-dependent manner and that circulating B4-scFv-fH localizes to damaged cochlear tissue in smoke-exposed ARHL mice. In Aim 2, I will test the hypothesis that local delivery of AAV5-pC3-CR2-fH and cochlear targeting of B4-scFv-fH can mitigate AMD pathology and prevent cochlear damage, respectively, in part by reducing macrophage activation. In Aim 3, as a postdoctoral trainee, I aspire to train in a lab with exper- tise in the aged human visual and auditory systems to expand my translational research in sensory loss. Over- all, my goal for this proposal is to demonstrate that AP inhibition in dual sensory loss can ameliorate pathology in part by reducing macrophage activation and reducing the proinflammatory microenvironment. My long-term goal is to expand my research and research training in translational sensory neuroscience.

项目摘要/摘要双重感觉丧失定义为与年龄相关的视力丧失的组合，例如与年龄相关的黄斑degenera- 年龄> 65岁的人（ARHL）（AMD）和与年龄相关的听力损失（ARHL）。 AMD，干和湿的形式，和ARHL是共享病因（例如吸烟和补体失调）的多因素疾病。重要的是，补体替代途径（AP）的放大环对控制至关重要，因为它是重新的大多数补体激活在细胞表面和细胞外膜上。 AP是被循环补体蛋白因子H（FH）抑制。补体是行动的一种机制之一 vated是在大坝上与新皮特结合或与损伤相关的分子模式结合的天然抗体（NABS）响应损伤的老化组织。补体效应子分子和NABS激活巨噬细胞，其他免疫细胞，增强可能导致神经变性的炎症状态。两个研究了融合蛋白CR2-FH和B4-SCFV-FH将用于靶向FH的抑制结构域到组织部位损害。补体受体2（CR2）结构域与补体碎片沉积位点结合，并结合单链抗体B4（B4-SCFV）结构域与在受损组织受损的组织上暴露的修饰膜联蛋白IV结合。两个都 CR2-FH和B4-SCFV-FH已显示在神经退行性疾病的小鼠模型中有效当系统地给药，或用于局部和通过基因治疗的CR2-FH时。感兴趣的是补充和在湿AMD小鼠模型的视网膜下空间和耳蜗组织中，巨噬细胞激活升高老年小鼠。另外，烟雾诱导的促炎，神经退行性AP激活和异常巨噬细胞活性已被视为眼睛和耳蜗。基于这些观察，我们假设向量驱动的CR2-FH并注入了B4-SCFV-FH将FH定位于受损的神经组织，并减轻湿AMD和 ARHL病理学，部分是通过减少巨噬细胞的激活，从而减少补体 - 巨噬细胞炎症反馈回路。我将以两个具体的目的检验该假设，结果将过渡到我的博士后研究。 AIM 1将提供C3启动子的概念证明一种依赖性的方式，并且循环的B4-SCFV-FH定位于损坏的人工耳蜗组织暴露于烟雾的ARHL小鼠。在AIM 2中，我将检验以下假设：局部交付AAV5-PC3-CR2-FH和 B4-SCFV-FH的耳蜗靶向可以减轻AMD病理，并在部分通过减少巨噬细胞激活。在AIM 3中，作为博士后实习生，我渴望在实验中训练在老年人的视觉和听觉系统中，可以扩大我在感官丧失方面的翻译研究。超过- 所有这些提案的目标是证明双重感觉丧失中的AP抑制可以改善病理部分是通过减少巨噬细胞激活并减少促炎的微环境。我的长期目标是扩大我在转化感官神经科学方面的研究和研究培训。