Apoptosis signal-regulating kinase 1 inhibition attenuates human airway smooth muscle growth and migration in chronic obstructive pulmonary disease.
Apoptosis signal-regulating kinase 1 inhibition attenuates human airway smooth muscle growth and migration in chronic obstructive pulmonary disease.
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DOI：
10.1042/cs20180398
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发表时间：
2018-07-31
期刊：
影响因子：
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通讯作者：
Sharma P
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作者：
Eapen MS;Kota A;Vindin H;McAlinden KD;Xenaki D;Oliver BG;Deshpande DA;Sohal SS;Sharma P
慢性阻塞性肺疾病（COPD）患者气道平滑肌（ASM）质量增加与疾病严重程度相关，并对这些患者的肺功能产生负面影响。因此，寻找针对COPD气道重塑和疾病进展的新疗法的临床需求显然尚未得到满足。凋亡信号调节激酶1 （Apoptosis signal- regulatory kinase 1, ASK1）是一种普遍表达的丝裂原活化蛋白激酶激酶激酶（MAP3K），可被各种应激刺激激活，包括活性氧（ROS）、肿瘤坏死因子(TNF)-α和脂多糖（LPS），已知可调节细胞增殖。慢性阻塞性肺病患者的ASM细胞在体外对有丝分裂原具有超增殖作用。然而，ASK1在ASM生长中的作用尚未确定。在这里，我们的目的是利用COPD患者的ASM细胞来确定ASK1抑制对ASM生长和促有丝分裂信号的影响。我们发现，与非COPD相比，ASK1在COPD肺asm束中的表达更高。用高选择性ASK1抑制剂TCASK10预处理ASM细胞，CyQuant检测结果显示，有丝分裂原（FBS、PDGF和EGF； 72小时）诱导的ASM生长呈剂量依赖性减少。此外，在ASM细胞中使用siRNA靶向ASK1可以阻止丝裂原诱导的细胞生长。此外，在抗有丝分裂潜能方面，ASK1抑制剂还能在体外阻止tgf β1诱导的ASM细胞迁移。免疫印迹显示，抗有丝分裂作用是由JNK和p38MAP激酶信号通路介导的，下游效应物JNK1/2和p38MAP激酶的磷酸化分别降低，而对ERK1/2没有影响。总的来说，这些发现确定了ASK1抑制的抗有丝分裂作用，并确定了一种新的途径，可以靶向减少或预防COPD中过度的ASM质量。
Increased airway smooth muscle (ASM) mass is observed in chronic obstructive pulmonary disease (COPD) which is correlated with disease severity and negatively impact lung function in these patients. Thus, there is clear unmet clinical need for finding new therapies which can target airway remodeling and disease progression in COPD. Apoptosis signal-regulating kinase 1 (ASK1) is a ubiquitously expressed mitogen-activated protein kinase kinase kinase (MAP3K) activated by various stress stimuli, including reactive oxygen species (ROS), tumor necrosis factor (TNF)-α, and lipopolysaccharide (LPS) and is known to regulate cell proliferation. ASM cells from COPD patients are hyper-proliferative to mitogens in vitro. However, the role of ASK1 in ASM growth is not established. Here, we aim to determine the effects of ASK1 inhibition on ASM growth and pro-mitogenic signaling using ASM cells from COPD patients. We found greater expression of ASK1 in ASM-bundles of COPD lung when compared with non-COPD. Pre-treatment of ASM cells with highly selective ASK1 inhibitor, TCASK10 resulted in a dose-dependent reduction in mitogen (FBS, PDGF and EGF; 72 hours)-induced ASM growth as measured by CyQuant assay. Further, molecular targeting of ASK1 using siRNA in ASM cells prevented mitogen-induced cell growth. In addition, to anti-mitogenic potential, ASK1 inhibitor also prevented TGFβ1-induced migration of ASM cells in vitro. Immunoblotting revealed that anti-mitogenic effects are mediated by JNK and p38MAP kinase-signaling pathways as evident by reduced phosphorylation of downstream effectors JNK1/2 and p38MAP kinases respectively with no effect on ERK1/2. Collectively, these findings establish the anti-mitogenic effect of ASK1 inhibition and identify a novel pathway that can be targeted to reduce or prevent excessive ASM mass in COPD.