Polarization of Human Macrophages by Interleukin-4 Does Not Require ATP-Citrate Lyase
Polarization of Human Macrophages by Interleukin-4 Does Not Require ATP-Citrate Lyase
复制标题
DOI：
10.3389/fimmu.2018.02858
复制
发表时间：
2018-12-04
影响因子：
7.3
通讯作者：
Bruene, Bernhard
中科院分区：
文献类型：
--
作者：
Namgaladze, Dmitry;Zukunft, Sven;Bruene, Bernhard
巨噬细胞暴露于Th2细胞因子白细胞介素（IL） IL-4和IL-13表现出明显的转录反应，通常被称为M2极化。最近，il -4诱导的小鼠骨髓源性巨噬细胞（bmdm）极化通过胞浆乙酰辅酶a生成酶atp -柠檬酸裂解酶（ACLY）的活性与乙酰辅酶a水平相关。在这里，我们研究了ACLY如何调节人单核细胞源性巨噬细胞（MDMs）中il -4刺激的基因表达。虽然多种ACLY抑制剂能减弱il -4诱导的靶基因表达，但这种效果不能通过沉默ACLY表达来重现。此外，ACLY抑制未能改变细胞乙酰辅酶a水平和组蛋白乙酰化。我们使用CRISPR/Cas9技术生成ACLY敲除的人THP-1巨噬细胞。虽然这些细胞表现出组蛋白乙酰化水平降低，但il -4诱导的基因表达保持完整。引人注目的是，ACLY抑制剂仍然抑制IL-4在ACLY敲除细胞中诱导靶基因，提示这些药物的脱靶作用。我们的研究结果表明，ACLY可能不是人巨噬细胞核质乙酰辅酶a和il -4诱导的极化的主要调节剂。此外，在解释ACLY药理抑制对基因表达的影响时应谨慎。
Macrophages exposed to the Th2 cytokines interleukin (IL) IL-4 and IL-13 exhibit a distinct transcriptional response, commonly referred to as M2 polarization. Recently, IL-4-induced polarization of murine bone marrow-derived macrophages (BMDMs) has been linked to acetyl-CoA levels through the activity of the cytosolic acetyl-CoA-generating enzyme ATP-citrate lyase (ACLY). Here, we studied how ACLY regulated IL-4-stimulated gene expression in human monocyte-derived macrophages (MDMs). Although multiple ACLY inhibitors attenuated IL-4-induced target gene expression, this effect could not be recapitulated by silencing ACLY expression. Furthermore, ACLY inhibition failed to alter cellular acetyl-CoA levels and histone acetylation. We generated ACLY knockout human THP-1 macrophages using CRISPR/Cas9 technology. While these cells exhibited reduced histone acetylation levels, IL-4-induced gene expression remained intact. Strikingly, ACLY inhibitors still suppressed induction of target genes by IL-4 in ACLY knockout cells, suggesting off-target effects of these drugs. Our findings suggest that ACLY may not be the major regulator of nucleocytoplasmic acetyl-CoA and IL-4-induced polarization in human macrophages. Furthermore, caution should be warranted in interpreting the impact of pharmacological inhibition of ACLY on gene expression.