The role of beta2-adrenergic receptor and interleukin-6 signaling in macrophage-driven choroidal neovascularization

项目总结： 候选人和职业发展计划：拉文博士是一名玻璃体视网膜外科医生、内科医生兼科学家， 西北大学眼科学助理教授。拉文博士拥有一个成功的博士学位， 雄厚的科学基础，并成功地平衡了临床职责和生产性的基础科学研究 在眼科进行了近8年的临床培训。他的长期职业目标是识别新的细胞和 先天免疫系统的分子靶点用于抗血管生成治疗以移动玻璃体视网膜 专家们越过了抗血管内皮生长因子(VEGF)时代，进入了个性化药物领域。至 要做到这一点，他的直接目标是发展他的职业生涯，成为一名有资金的、独立的内科科学家， 在炎症和血管生成方面的专业知识。K08奖将通过以下方式帮助实现这些目标 发展拉文博士在高级免疫学和生物信息学方面的科学和专业技能。拉文博士 他的合作导师，帕尔曼博士和夸金博士已经制定了实现这些目标的详细战略 目标通过精心计划的课程作业、教学方法、实验室技术和合作 西北大学。目前的建议将为未来R01级关于 脉络膜新生血管(CNV)中先天免疫和血管生成的交集。 研究计划：新生血管性老年性黄斑变性(NAMD)是老年人致盲的主要原因 在发达国家，仅通过抑制血管内皮生长因子进行治疗。尽管非常有效，15%的患者仍然失去了 视觉，尽管最大限度地抗血管内皮生长因子治疗。有证据表明促血管生成的巨噬细胞在NAMD中的作用 发病机制，但目前还没有针对这些细胞的治疗方法。白介素6(IL-6)是一种促进剂。 已知的炎性细胞因子由巨噬细胞产生，与NAMD活动相关，是必要的 对于激光诱导的CNV，建立NAMD的小鼠模型。我们的初步数据表明，β2-肾上腺素能 受体(AR)抑制通过降低巨噬细胞中IL-6的水平来减少激光诱导的CNV面积。我们的中央 假说是β2-AR信号影响巨噬细胞分化，促进促血管生成 巨噬细胞表型，增加IL-6水平，并通过直接激活IL-6受体激活血管生成 在内皮细胞上增加CNV面积。为了验证这一假设，我们制定了以下具体目标： 1)确定视网膜/脉络膜细胞类型(S)在β2-AR拮抗作用中表达IL-6的反应 2)确定产生IL-6的细胞类型和对IL-6有反应的细胞类型，以增加激光诱导的CNV。 3)阐明β2-AR和IL-6缺陷对CNV过程中巨噬细胞转录调控的影响。 这些目标的完成将决定对β2-AR拮抗反应的细胞类型，产生IL-6，以及 应用IL-6增加新生血管面积。这些数据将为NAMD的抗IL-6治疗奠定基础。此外， 我们将描述β2-AR抑制如何影响CNV环境中巨噬细胞的转录图谱， 这将确定新的抗炎和抗血管生成治疗靶点。

PROJECT SUMMARY: Candidate and Career Development Plan: Dr. Lavine is a vitreoretinal surgeon, physician-scientist, and Assistant Professor of Ophthalmology at Northwestern University. Dr. Lavine had a successful PhD, laying his strong scientific foundation, and has successfully balanced clinical duties with productive, basic science research in ophthalmology for the past 8 years of clinical training. His long-term career goal is to identify novel cellular and molecular targets of the innate immune system for anti-angiogenic therapy in order to move vitreoretinal specialists past the anti-vascular endothelial growth factor (VEGF) era and into individualized medicine. To accomplish this, his immediate goal is to develop his career as a funded, independent physician-scientist with expertise in the intersection of inflammation and angiogenesis. This K08 award will help achieve these goals by developing Dr. Lavine’s scientific and professional skills in advanced immunology and bio-informatics. Dr. Lavine and his co-mentors, Dr. Perlman and Dr. Quaggin, have developed a detailed strategy to achieve these objectives through carefully planned course work, didactics, laboratory techniques, and collaborations at Northwestern University. The current proposal will lay the foundation for future R01-level proposals on the intersection between innate immunity and angiogenesis in choroidal neovascularization (CNV). Research Plan: Neovascular age-related macular degeneration (nAMD) is the leading cause of blindness in the developed world, and is treated solely by inhibiting VEGF. Although highly effective, 15% of patients still lose vision despite maximal anti-VEGF therapy. Evidence suggests a role for pro-angiogenic macrophages in nAMD pathogenesis, but currently there are no therapies to specifically target these cells. Interleukin-6 (IL-6), a pro- inflammatory cytokine known to be produced by macrophages, correlates with nAMD activity and is necessary for laser-induced CNV, a mouse model of nAMD. Our preliminary data demonstrate that beta2-adrenergic receptor (AR) inhibition decreases laser-induced CNV area by reducing IL-6 levels in macrophages. Our central hypothesis is that beta2-AR signaling influences macrophage differentiation, promotes a pro-angiogenic macrophage phenotype, increases IL-6 levels, and activates angiogenesis by activating the IL-6 receptor directly on endothelial cells to increase CNV area. To test this hypothesis, we formulated the following specific aims: 1) Determine the retinal/choroidal cell type(s) that express interleukin-6 in response to beta2-AR antagonism 2) Identify the cell type that produces IL-6 and the cell type that responds to IL-6 to increase laser-induced CNV. 3) Delineate how beta2-AR and IL-6 deficiency regulate the transcriptional profile of macrophages during CNV. Completion of these aims will determine the cell types that respond to beta2-AR antagonism, produce IL-6, and respond to IL-6 to increase CNV area. These data will set the stage for anti-IL-6 therapy for nAMD. Furthermore, we will delineate how beta2-AR inhibition influences the transcriptomic profile of macrophages in the CNV milieu, which will identify new anti-inflammatory and anti-angiogenic therapeutic targets.