Oxidized Phospholipids Promote NETosis and Arterial Thrombosis in LNK(SH2B3) Deficiency.
Oxidized Phospholipids Promote NETosis and Arterial Thrombosis in LNK(SH2B3) Deficiency.
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DOI：
10.1161/circulationaha.121.056414
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发表时间：
2021-12-14
期刊：
影响因子：
37.8
通讯作者：
Tall AR
中科院分区：
文献类型：
--
作者：
Dou H;Kotini A;Liu W;Fidler T;Endo-Umeda K;Sun X;Olszewska M;Xiao T;Abramowicz S;Yalcinkaya M;Hardaway B;Tsimikas S;Que X;Bick A;Emdin C;Natarajan P;Papapetrou EP;Witztum JL;Wang N;Tall AR
补充数字内容可在文本中找到。LNK/SH 2B 3通过造血细胞因子受体抑制Janus激酶/信号转导和转录激活因子（JAK/STAT）信号传导。全基因组关联研究显示LNK中常见的单核苷酸多态性（R262 W，T等位基因）与嗜中性粒细胞增多症、血小板增多症和冠状动脉疾病相关。我们已经证明LNK（TT）降低LNK功能，LNK缺陷小鼠显示出显著的血小板-中性粒细胞聚集，加速动脉粥样硬化和血栓形成。血小板-中性粒细胞相互作用可促进中性粒细胞胞外陷阱（NET）形成。本研究的目的是评估NETs在造血Lnk缺陷小鼠动脉粥样硬化和血栓形成中的作用。我们培育了Lnk和NETosis必需酶PAD 4（肽基精氨酸脱亚胺酶4）联合缺乏的小鼠，并将其骨髓移植到Ldlr-/-小鼠中。我们评估了LNK在人类和携带JAK 2（JAK 2 V617 F）功能获得变体的小鼠中动脉粥样硬化血栓形成中的作用。Lnk缺陷小鼠表现出加速的颈动脉血栓形成，具有显著的NETosis，其被PAD 4缺陷完全逆转。与野生型血小板或野生型中性粒细胞相比，凝血酶激活的Lnk-/-血小板与Lnk-/-中性粒细胞孵育时促进NETosis增加。这涉及增加表面暴露和从Lnk-/-血小板释放氧化磷脂（OxPL），以及增加Lnk-/-中性粒细胞对OxPL的引发和应答。为了抵消OxPL的影响，我们引入了表达E06的单链可变区片段（E06-scFv）的转基因。E06-scFv逆转Lnk-/-小鼠中加速的NETosis、动脉粥样硬化和血栓形成。我们还显示，当人类诱导多能干细胞来源的LNK（TT）中性粒细胞与LNK（TT）血小板/巨核细胞孵育时，NETosis增加，但在同基因LNK（CC）对照中则没有，证实了人类相关性。使用英国生物银行的数据，我们发现JAK 2 VF突变的个体只有在携带LNK R262 W等位基因时才显示出冠心病风险增加。与Jak 2 VFLnk +/+对照组相比，具有造血Lnk+/-和Jak 2 VF克隆造血的小鼠显示加速的动脉血栓形成，但不显示动脉粥样硬化。造血Lnk缺乏以OxPL依赖性方式促进NETosis和动脉血栓形成。LNK（R262 W）降低人类血小板和中性粒细胞中的LNK功能，促进NETosis，并增加携带Jak 2 VF突变的人患冠状动脉疾病的风险。靶向OxPL的治疗可能有益于遗传定义的人群中的冠状动脉疾病。
Supplemental Digital Content is available in the text. LNK/SH2B3 inhibits Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling by hematopoietic cytokine receptors. Genome-wide association studies have shown association of a common single nucleotide polymorphism in LNK (R262W, T allele) with neutrophilia, thrombocytosis, and coronary artery disease. We have shown that LNK(TT) reduces LNK function and that LNK-deficient mice display prominent platelet–neutrophil aggregates, accelerated atherosclerosis, and thrombosis. Platelet–neutrophil interactions can promote neutrophil extracellular trap (NET) formation. The goals of this study were to assess the role of NETs in atherosclerosis and thrombosis in mice with hematopoietic Lnk deficiency. We bred mice with combined deficiency of Lnk and the NETosis-essential enzyme PAD4 (peptidyl arginine deiminase 4) and transplanted their bone marrow into Ldlr–/– mice. We evaluated the role of LNK in atherothrombosis in humans and mice bearing a gain of function variant in JAK2 (JAK2V617F). Lnk-deficient mice displayed accelerated carotid artery thrombosis with prominent NETosis that was completely reversed by PAD4 deficiency. Thrombin-activated Lnk–/– platelets promoted increased NETosis when incubated with Lnk–/– neutrophils compared with wild-type platelets or wild-type neutrophils. This involved increased surface exposure and release of oxidized phospholipids (OxPL) from Lnk–/– platelets, as well as increased priming and response of Lnk–/– neutrophils to OxPL. To counteract the effects of OxPL, we introduced a transgene expressing the single-chain variable fragment of E06 (E06-scFv). E06-scFv reversed accelerated NETosis, atherosclerosis, and thrombosis in Lnk–/– mice. We also showed increased NETosis when human induced pluripotent stem cell–derived LNK(TT) neutrophils were incubated with LNK(TT) platelet/megakaryocytes, but not in isogenic LNK(CC) controls, confirming human relevance. Using data from the UK Biobank, we found that individuals with the JAK2VF mutation only showed increased risk of coronary artery disease when also carrying the LNK R262W allele. Mice with hematopoietic Lnk+/– and Jak2VF clonal hematopoiesis showed accelerated arterial thrombosis but not atherosclerosis compared with Jak2VFLnk+/+ controls. Hematopoietic Lnk deficiency promotes NETosis and arterial thrombosis in an OxPL-dependent fashion. LNK(R262W) reduces LNK function in human platelets and neutrophils, promoting NETosis, and increases coronary artery disease risk in humans carrying Jak2VF mutations. Therapies targeting OxPL may be beneficial for coronary artery disease in genetically defined human populations.