Leukocyte NADPH Oxidase Variants in Lupus

该项目将研究超氧化物生成中的变体之间的联系， 白细胞NADPH氧化酶2（NOX 2）、遗传易感性和SLE发病机制 我们激动人心的发现将白细胞NADPH氧化酶NCF 2 H389 Q和R395 W 人类SLE的突变。我们的中心假设是SLE不是一个单一的疾病实体， 而是代表基于某些遗传风险因素组合的不同亚型。 因此，我们假设狼疮与NADPH氧化酶活性降低有关， NOX 2功能变异体代表SLE的一种独特亚型，其具有特定的调节途径， 发病机制，独特的生物标志物，因此对特定疗法有反应。 在本研究中，我们将确定功能重要的NADPH的总体患病率， 使用下一代测序（NGS）技术， 检测并验证9个NADPH氧化酶基因中的重要新变异， NADPH氧化酶复合物对人类SLE遗传易感性的贡献 (aim 1）并将进一步分析鉴定的和功能上有前途的H389 Q和R395 W NCF 2变体（分目标1.3和1.4）。在具体目标2中，我们将研究 NADPH氧化酶活性降低促进狼疮发病机制的实验研究 具有NADPH氧化酶遗传缺陷（包括部分缺陷）的狼疮模型 （单倍不足NCF 2，亚型NCF 2）和NADPH氧化酶缺失的选择性 骨髓谱系这些机制分析将在几个层面上进行：基因表达 研究（子目标2.1），以确定生物标志物和调节NOX 2相关通路 狼疮; I型干扰素在驱动发病机制中的作用（子目标2.2）;不同细胞亚群在 驱动过程（子目标2.3）; NETosis在发病机制中的作用（子目标2.4），最后 评估潜在的NOX 2激动剂作为该狼疮亚组的可能特异性治疗方式 （次级目标2.5）。

This project will examine the connection between variants in the superoxide-generating leukocyte NADPH oxidase 2 (NOX2), genetic susceptibility and pathogenesis of SLE to follow up on our exciting discovery linking leukocyte NADPH oxidase NCF2 H389Q and R395W mutations to human SLE. Our central hypothesis is that SLE is not a single disease entity, but rather represents distinct subtypes based on combination of some of the genetic risk factors. Thus, we hypothesize that lupus linked to reduced NADPH oxidase activity as a consequence of NOX2 functional variants represent a distinct subtype of SLE with specific pathways regulating pathogenesis, unique biomarkers and therefore responsive to specific therapies. In the present study we will determine the overall prevalence of functionally important NADPH oxidase variants in lupus subjects using next generation sequencing (NGS) technologies to detect and validate important new variants in 9 NADPH oxidase genes and determine the extent of the contribution of the NADPH oxidase complex to the genetic predisposition to human SLE (aim 1) and will further analyze the identified and functionally promising H389Q and R395W NCF2 variants (subaims 1.3 and 1.4). In specific aim 2 we will investigate mechanisms by which reduced NADPH oxidase activity promotes lupus pathogenesis using newly created mouse models of lupus with genetic defects in the NADPH oxidase, including partial deficiency (haploinsufficient NCF2, hypomorphic NCF2) and with deletion of NADPH oxidase in selected myeloid lineages. These mechanistic analyses will be done on several levels: gene expression studies (subaim 2.1) to define biological markers and pathways regulating NOX2 associated lupus; role of type I IFNs in driving pathogenesis (subaim 2.2); role of different cell subsets in driving the process (subaim 2.3); role of NETosis in the pathogenesis (subaim 2.4) and finally assessing potential NOX2 agonists as likely specific treatment modality to this subset of lupus (subaim 2.5).