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：
10393958
负责人：
Nathaniel Bryan David Parsons
金额：
$ 4.5万
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/10393958
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 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/antagonist 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.

项目概要/摘要双重感觉丧失被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变性 (AMD) 和年龄相关性听力损失 (ARHL) 在年龄 > 65 岁的人群中同时发生。 AMD，干式和湿式， ARHL 和 ARHL 是多因素疾病，其病因相同，例如吸烟和补体失调。重要的是，补体旁路途径 (AP) 的放大环对于控制至关重要，因为它是重新负责细胞表面和细胞外膜上的大部分补体激活。美联社是被循环补体蛋白 H 因子 (fH) 抑制。补体激活的机制之一 vated 是与新表位或损伤相关分子模式结合的天然抗体 (nAb) 因损伤而老化的组织。补体效应分子和 nAb 激活巨噬细胞并其他免疫细胞，加剧炎症状态，从而导致神经退行性变。两个好好研究一下融合蛋白 CR2-fH 和 B4-scFv-fH 将用于将 fH 的抑制结构域靶向组织部位损害。补体受体 2 (CR2) 结构域与补体片段沉积位点结合，并且单链抗体 B4 (B4-scFv) 结构域与暴露在受损组织上的修饰膜联蛋白 IV 结合。两个都 CR2-fH 和 B4-scFv-fH 已被证明在神经退行性疾病小鼠模型中有效当全身给药时，或通过基因治疗局部给药 CR2-fH 时。有趣的是补充和湿 AMD 小鼠模型的视网膜下腔和耳蜗组织中的巨噬细胞活化升高年老的老鼠。此外，烟雾诱发的促炎、神经退行性 AP 激活和异常巨噬细胞的活动，已见于眼睛和耳蜗。根据这些观察，我们假设载体驱动的 CR2-fH 和注射的 B4-scFv-fH 将 fH 定位于受损的神经组织并减轻湿性 AMD 和 ARHL 病理学，部分是通过减少巨噬细胞活化，从而减少补体- 巨噬细胞炎症反馈环路。我将在两个具体目标中检验这个假设，其结果将过渡到我的博士后研究。目标 1 将提供 C3 启动子被调节的概念证明补体依赖性方式，并且循环 B4-scFv-fH 定位于受损的耳蜗组织暴露于烟雾的 ARHL 小鼠。在目标 2 中，我将测试 AAV5-pC3-CR2-fH 的本地递送和 B4-scFv-fH 的耳蜗靶向可以分别减轻 AMD 病理并预防耳蜗损伤部分通过减少巨噬细胞活化来实现。在目标 3 中，作为一名博士后实习生，我渴望在拥有专家的实验室中进行培训结合老年人的视觉和听觉系统来扩展我在感觉丧失方面的转化研究。超过- 总之，我这个提案的目标是证明双感觉丧失中的 AP 抑制可以改善病理学部分是通过减少巨噬细胞活化和减少促炎微环境来实现的。我的长期目标是扩大我在转化感觉神经科学方面的研究和研究培训。