Monocyte and macrophage polarization in systemic juvenile idiopathic arthritis and macrophage activation syndrome.

项目总结/摘要 系统性幼年特发性关节炎是一种严重的儿童炎症性疾病， 致命并发症的风险，包括巨噬细胞活化综合征（MAS）。的表型和功能 系统性JIA中的关键效应单核细胞和巨噬细胞仍不清楚。这些细胞采用不同的 基于特定激活信号的极化状态，由microRNA修饰以"微调"这些 转录反应。系统性JIA患者的循环单核细胞似乎显示出独特的混合极化 表型，并且关于组织驻留的巨噬细胞如何在生长期间进一步改变知之甚少。 MAS的出现。值得注意的是，尽管有有效的治疗，全身性JIA儿童仍然有MAS的风险。 因此，迫切需要表征系统性炎症中单核细胞和组织巨噬细胞的表型。 JIA，特别是它们如何有助于MAS的出现。由于缺乏这种知识， 有效预防和治疗MAS的新策略仍将是一个艰巨的挑战。在这方面的目标 应用是定义极化特异性基因表达特征和microRNA的功能作用， 单核细胞和巨噬细胞从儿童全身JIA和小鼠模型全身JIA/MAS。我们 中心假设是来自全身性JIA儿童的单核细胞具有独特的极化模式， 受microRNA调节，并在MAS期间进一步失调。本申请书所述的研究计划 有三个特定的目的：定义外周血中极化特异性和单细胞基因表达谱， 单核细胞和骨髓巨噬细胞从全身JIA和MAS患者（目的1），以确定具体的 microRNA靶向的分子通路，对于单核细胞和巨噬细胞极化是必需的， 这些疾病（目的2），并确定基因表达特征和microRNA介导的功能， 极化的骨髓群体在MAS的小鼠模型中（Aim 3）。这些独立和互补的 aims将利用microRNA表达谱和单细胞RNAseq等新兴技术， 确定单核细胞和巨噬细胞中极化特异性转录特征的分子控制 全身性JIA和MAS患儿。这些研究结果预计将对人类的健康产生积极的影响。 通过提供新的靶点来调节全身性JIA的炎症并影响MAS的风险。 Grant Schulert博士目前是辛辛那提风湿病学分部的儿科讲师 儿童医院医疗中心。Schulert博士的长期目标是发展独立的研究事业 作为一名儿科风湿病学家，他专注于炎症的发病机制和新的治疗方法， 紊乱在完成这些拟议的目标，Schulert博士将利用超过10年的经验， 经验研究炎症和先天免疫，但也将获得新的技能和知识的领域 功能基因组学、microRNA生物学和炎症性疾病的动物模型。Schulert博士及其 主要导师Alexei Grom博士制定了全面的职业发展计划，其中包括四个关键因素 组成部分：实验室实验;指导监督指导他的指导委员会;领导 通过CCHMC领导基础计划和其他研讨会进行技能培养; 通过科学会议、校内会议和CCHMC "K俱乐部"促进专业发展。 他的职业发展将由一个由Grom博士主持的跨学科导师委员会指导。 Schulert博士的导师委员会的其他成员将促进新技能的获得， 知识在整个奖励期间，同时作为拟议的研究目标的主要合作者。 CCHMC的环境非常支持Schulert博士在这个关键的早期阶段的发展。 他的研究生涯。这个机构提供了各种资源，包括物质和智力，以支持他的工作。 职业发展，CCHMC已承诺保护至少75%的时间用于研究期间， 奖励期。CCHMC一直是首屈一指的儿科培训和研究计划的所在地 几十年来，该部门的研究组合是相当多样化的，从基因组， 从幼年特发性关节炎（JIA）到发现儿童期发病狼疮的器官特异性生物标志物， 新药靶点的国际临床试验。 总的来说，在指导委员会的帮助下， 出色的机构环境，包括辛辛那提儿童医院的共享核心设施 医学中心，将提供理想的环境，为博士Schulert实现他的职业目标，成为一个 儿科流变学的独立研究者。

PROJECT SUMMARY / ABSTRACT Systemic juvenile idiopathic arthritis is a severe inflammatory disease of childhood conferring significant risk for fatal complications including Macrophage Activation Syndrome (MAS). The phenotype and function of the key effector monocytes and macrophages in systemic JIA remains unclear. These cells adopt distinct polarization states based on specific activating signals, modified by microRNA to “fine-tune” these transcriptional responses. Circulating monocytes in systemic JIA appear to display a unique mixed polarization phenotype, and little is known regarding how tissue resident macrophages are further altered during emergence of MAS. Notably, despite effective treatment children with systemic JIA remain at risk for MAS. Thus, there is a critical need to characterize the phenotype of monocytes and tissue macrophages in systemic JIA, and particularly how they contribute to emergence of MAS. In the absence of such knowledge, developing novel strategies to effectively prevent and treat MAS will remain a formidable challenge. The objective in this application is to define polarization-specific gene expression signatures and functional roles of microRNA in monocytes and macrophages from children with systemic JIA and a mouse model of systemic JIA/MAS. Our central hypothesis is monocytes from children with systemic JIA have a distinct polarized pattern which is regulated by microRNA, and further dysregulated during MAS. The research plan described in this application has three Specific Aims: to define polarization-specific and single-cell gene expression profiles in peripheral monocytes and bone marrow macrophages from systemic JIA and MAS patients (Aim 1), to identify specific molecular pathways targeted by microRNA that are essential for monocyte and macrophage polarization in these disorders (Aim 2), and to determine gene expression signatures and microRNA-mediated functions of polarized myeloid populations in a mouse model of MAS (Aim 3). These independent and complementary aims will utilize emerging technologies such as microRNA expression profiling and single-cell RNAseq to define the molecular control of polarization-specific transcriptional signatures in monocytes and macrophages in children with systemic JIA and MAS. Together these findings are expected to have positive translational impact, through providing new targets to modulate inflammation in systemic JIA and impact the risk for MAS. Dr. Grant Schulert is currently an Instructor of Pediatrics in the Division of Rheumatology at Cincinnati Children's Hospital Medical Center. Dr. Schulert's long-term goal is to develop an independent research career as a pediatric rheumatologist focused on the pathogenesis and novel treatment approaches for inflammatory disorders. In the completion of these proposed aims, Dr. Schulert will draw upon greater than 10 years' experience studying inflammation and innate immunity, but will also gain new skills and knowledge in the areas of functional genomics, microRNA biology and animal models of inflammatory disorders. Dr. Schulert and his primary mentor, Dr. Alexei Grom, have developed a comprehensive career development plan with four key components: laboratory experimentation; mentored oversight guided by his mentorship committee; leadership and skills building through the CCHMC Leadership Foundations Program and other workshops; and academic and professional development through scientific meetings, intramural conferences and the CCHMC “K Club”. His career development will be guided by an interdisciplinary mentorship committee chaired by Dr. Grom. Additional members of Dr. Schulert's mentorship committee will facilitate the acquisition of new skills and knowledge throughout the award period, while serving as key collaborators for the proposed research aims. The environment at CCHMC is highly supportive of Dr. Schulert's development in this key early stage of his research career. This institution offers a variety of resources both physical and intellectual to support his career development, and CCHMC has committed to protecting at least 75% of his time for research during the award period. CCHMC has been the home of the premier training and research program in pediatric rheumatology for several decades. The divisional research portfolio is quite diverse, spanning from genomic in juvenile idiopathic arthritis (JIA) to the discovery of organ-specific biomarkers for childhood-onset lupus to international clinical trials of new drug targets. Collectively, completion of the proposed studies, aided by the mentoring committee and with the outstanding institutional environment including shared core facilities available at Cincinnati Children's Hospital Medical Center, will provide the ideal environment for Dr. Schulert to achieve his career goal of becoming an independent investigator within Pediatric Rheumatology.