Kwon Yoojung, Choi Yunji, Kim Misun, Jo Hyein, Jeong Myeong Seon, Jung Hyun Suk, Jeoung Dooil
Department of Biochemistry, Kangwon National University, Chuncheon 24341, South Korea.
KM Science Research Division, Korea Institute of Oriental Medicine, Daejeon 34054, South Korea.
Mol Immunol. 2024 Feb;166:1-15. doi: 10.1016/j.molimm.2023.12.007. Epub 2024 Jan 3.
Histone deacetylase 6 (HDAC6) has been shown to play an important role in allergic inflammation. This study hypothesized that novel downstream targets of HDAC6 would mediate allergic inflammation. Experiments employing HDAC6 knock out C57BL/6 mice showed that HDAC6 mediated passive cutaneous anaphylaxis (PCA) and passive systemic anaphylaxis (PSA). Antigen stimulation increased expression of N-myc (MYCN) and CXCL3 in an HDAC6-dependent manner in the bone marrow-derived mast cells. MYCN and CXCL3 were necessary for both PCA and PSA. The role of early growth response 3 (EGR3) in the regulation of HDAC6 expression has been reported. ChIP assays showed EGR3 as a direct regulator of MYCN. miR-34a-5p was predicted to be a negative regulator of MYCN. Luciferase activity assays showed miR-34a-5p as a direct regulator of MYCN. miR-34a-5p mimic negatively regulated PCA and PSA. MYCN decreased miR-34a-5p expression in antigen-stimulated rat basophilic leukemia cells (RBL2H3). MYCN was shown to bind to the promoter sequence of CXCL3. In an IgE-independent manner, recombinant CXCL3 protein increased expression of HDAC6, MYCN, and β-hexosaminidase activity in RBL2H3 cells. Mouse recombinant CXCL3 protein enhanced the angiogenic potential of the culture medium of RBL2H3. CXCL3 was necessary for the enhanced angiogenic potential of the culture medium of antigen-stimulated RBL2H3. The culture medium of RBL2H3 was able to induce M2 macrophage polarization in a CXCL3-dependent manner. Recombinant CXCL3 protein also increased the expression of markers of M2 macrophage. Thus, the identification of the novel role of HDAC6-MYCN-CXCL3 axis can help better understand the pathogenesis of anaphylaxis.
组蛋白去乙酰化酶6（HDAC6）已被证明在过敏性炎症中起重要作用。本研究假设HDAC6的新下游靶点将介导过敏性炎症。使用HDAC6基因敲除的C57BL/6小鼠进行的实验表明，HDAC6介导被动皮肤过敏反应（PCA）和被动全身过敏反应（PSA）。抗原刺激以HDAC6依赖的方式增加骨髓来源肥大细胞中N-myc（MYCN）和CXCL3的表达。MYCN和CXCL3对于PCA和PSA都是必需的。早期生长反应3（EGR3）在HDAC6表达调控中的作用已有报道。染色质免疫沉淀分析显示EGR为MYCN的直接调节因子。miR-34a-5p被预测为MYCN的负调节因子。荧光素酶活性分析显示miR-34a-5p为MYCN的直接调节因子。miR-34a-5p模拟物对PCA和PSA起负调节作用。MYCN降低抗原刺激的大鼠嗜碱性白血病细胞（RBL2H3）中miR-34a-5p的表达。MYCN被证明与CXCL3的启动子序列结合。以不依赖IgE的方式，重组CXCL3蛋白增加RBL2H3细胞中HDAC6、MYCN的表达以及β-己糖胺酶活性。小鼠重组CXCL3蛋白增强了RBL2H3培养基的血管生成潜力。CXCL3是抗原刺激的RBL2H3培养基血管生成潜力增强所必需的。RBL2H3培养基能够以CXCL3依赖的方式诱导M2巨噬细胞极化。重组CXCL3蛋白也增加了M2巨噬细胞标志物的表达。因此，确定HDAC6-MYCN-CXCL3轴的新作用有助于更好地理解过敏反应的发病机制。
