Macrophages and Treatment-Resistant NVAMD

 描述（由申请人提供）：候选人是一名视网膜临床医生-科学家，其主要职业目标是通过（1）阐明疾病机制，特别关注巨噬细胞来促进年龄相关性黄斑变性（AMD）患者的护理;和（2）确定新的疾病生物标志物和治疗靶点。除了临床培训，他还完成了临床研究奖学金，并在部门K12职业发展计划中度过了两年。在第一年，他在各个实验室轮换，在第二年，他从事了一个新的项目，研究活化的巨噬细胞在新生血管重塑（NVR）中的作用，新生毛细血管转化为血管周围纤维化的分支小动脉，以及耐药性新生血管性AMD（NVAMD）的主要原因。他在当前提案中的直接职业发展目标是通过鉴定产生NVR促进巨噬细胞的特定血液单核细胞亚群和表征NVR环境中调节巨噬细胞效应器功能的机制来进一步了解NVR病理生物学。该候选人计划在第三年早些时候提交R 01申请，该申请涉及一个创新的翻译项目，以表征实验性CNV和NVAMD中巨噬细胞介导的NVR的生物学，将单核细胞亚群频率和激活状态与CNV形态学和治疗抗性NVAMD相关联。在免疫学，分子生物学，显微镜，部门研究研讨会和先进的负责任的研究行为将在奖励期间获得额外的教学培训，候选人将在国家会议上介绍他的研究结果，并提交他的工作出版。工作环境：顾问委员会的指导和专业知识，眼科和药理学/癌症生物学的杜克部门的广泛资源，以及重大的机构承诺将提供所需的支持，为这位候选人完成拟议的研究项目，并成功地过渡到一个独立的研究生涯。研究：虽然玻璃体内抗VEGF治疗改善了视力结果，但40-50%的患者表现出治疗抵抗性NVAMD / PDA。PDA在CNV形态反映NVR的患者中更常见。候选人和他的主要导师已经建立了巨噬细胞作为NVR的驱动因素，初步数据表明，低级别全身暴露于三种不同的病原体相关分子模式（PAMP）促进了激光诱导CNV小鼠模型中的NVR。尽管每种PAMP具有不同的活化受体，但已知所有三种PAMP活化的信号级联都动员钙并活化钙结合蛋白钙调蛋白。因此，初步数据表明，小鼠无钙/钙调蛋白依赖性蛋白激酶激酶2（CaMKK 2），一种已知在巨噬细胞中扩增PAMP激活的效应子途径的中间激酶，表现出降低的NVR。中心假设是PAMP刺激通过激活CaMKK 2介导的巨噬细胞效应子功能来调节NVR。具体而言，假设低级PAMP刺激（1）增加血液中循环非经典Ly 6Clo单核细胞的频率;（2）促进Ly 6Clo单核细胞衍生的巨噬细胞的活化，增加纤维化生长因子的表达，（3）通过激活CaMKK 2信号传导（图2）。纤维化因子募集并激活血管平滑肌细胞和肌成纤维细胞，导致NVR的发展。目的1将解决的假设，PAMP刺激的NVR在实验性新血管形成介导的活化的巨噬细胞产生的非经典的Ly 6Clo血液单核细胞。目的2将阐明CaMKK 2调节巨噬细胞介导的NVR的假设。

 DESCRIPTION (provided by applicant): The candidate is a retina clinician-scientist whose primary career goal is to advance the care of patients with age-related macular degeneration (AMD) by (1) elucidating disease mechanisms, with a specific focus on macrophages; and (2) identifying novel disease biomarkers and targets for therapy. In addition to clinical training, he has completed a clinical research fellowship and has spent two years in a Departmental K12 Career Development Program. In the first year, he rotated through various laboratories and in the second year, he has pursued a novel project investigating the role of activated macrophages in neovascular remodeling (NVR), the transformation of nascent capillaries into branching arterioles with perivascular fibrosis, and a major cause of treatment-resistant neovascular AMD (NVAMD). His immediate career development goals in the current proposal are to further understanding of NVR pathobiology by identifying the specific blood monocyte subset(s) that give rise to NVR-promoting macrophages and by characterizing mechanisms that regulate macrophage effector function in the setting of NVR. The candidate plans an R01 submission in early year three on an innovative translational project to characterize the biology of macrophage-mediated NVR in experimental CNV and NVAMD, correlating monocyte subset frequencies and activation states with CNV morphology and treatment-resistant NVAMD. Additional didactic training in immunology, molecular biology, microscopy, departmental research seminars, and advanced responsible conduct of research will be obtained during the award period, and the candidate will present his findings at national meetings and submit his work for publication. Environment: The mentorship and expertise of the advisory committee, the extensive resources of the Duke Departments of Ophthalmology and Pharmacology / Cancer Biology, and the significant institutional commitment will provide the support needed for this candidate to complete the proposed research project and transition successfully to an independent research career. Research: Although intravitreal anti-VEGF therapies have improved vision outcomes, 40-50% of patients exhibit treatment-resistant NVAMD / PDA. PDA occurs more frequently in patients with CNV morphology that reflects NVR. The candidate and his primary mentor have established macrophages as a driver of NVR, and preliminary data suggests that low-grade systemic exposure to three different pathogen-associated molecular patterns (PAMPs) promotes NVR in a murine model of laser-induced CNV. In spite of each PAMP having a distinct activating receptor, all three PAMP-activated signaling cascades are known to mobilize calcium and activate the calcium-binding protein, calmodulin. Accordingly, preliminary data suggests that mice null for calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2), an intermediate kinase known to amplify PAMP-activated effector pathways in macrophages, demonstrate reduced NVR. The central hypothesis is that PAMP-stimulation regulates NVR through activation of CaMKK2-mediated macrophage effector function. Specifically, low-grade PAMP stimulation is hypothesized to (1) increase the frequency of circulating nonclassical Ly6Clo monocytes in blood; and (2) promotes activation of Ly6Clo monocyte-derived macrophages, increasing expression of fibrogenic growth factors, (3) via activation of CaMKK2 signaling (Fig. 2). Fibrogenic factors recruit and activate vascular smooth muscle cells and myofibroblasts, leading to development of NVR. Aim 1 will address the hypothesis that PAMP-stimulated NVR in experimental neovascularization is mediated by activated macrophages arising from nonclassical Ly6Clo blood monocytes. Aim 2 will address the hypothesis that CaMKK2 regulates macrophage-mediated NVR.