Spinelli Amy M, Liu Yongfeng, Sun Li-Yan, González-Cobos José C, Backs Johannes, Trebak Mohamed, Singer Harold A
Center for Cardiovascular Sciences, Albany Medical College, Albany, NY, USA.
Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, PA, USA.
Pflugers Arch. 2015 Dec;467(12):2541-54. doi: 10.1007/s00424-015-1713-5. Epub 2015 Jun 20.
Airway smooth muscle (ASM) is a key target cell in allergen-induced asthma known to contribute to airway hyperresponsiveness (AHR) and chronic airway remodeling. Changes in ASM calcium homeostasis have been shown to contribute to AHR although the mechanisms and Ca(2+) signal effectors are incompletely understood. In the present study, we tested the function of ASM multifunctional protein kinase Ca(2+)/calmodulin-dependent kinase II (CaMKII) isoforms CaMKIIδ and CaMKIIγ in allergen-induced AHR and airway remodeling in vivo. Using a murine model of atopic asthma, we demonstrate that CaMKIIδ protein is upregulated in ASM derived from ovalbumin (OVA)-treated animals compared to controls. A genetic approach to conditionally knock out smooth muscle CaMKIIδ and CaMKIIγ in separate Cre-loxp systems was validated, and using this loss-of-function approach, the function of these CaMKII isoforms was tested in ovalbumin (OVA)-induced airway remodeling and AHR. OVA treatment in control mice had no effect on ASM remodeling in this model of AHR, and CaMKIIδ knockouts had no independent effects on ASM content. However, at 1 day post-final OVA challenge, OVA-induced AHR was eliminated in the CaMKIIδ knockouts. OVA-induced peribronchial inflammation and bronchoalveolar lavage fluid (BALF) levels of the Th2 cytokine IL-13 were significantly decreased in the CaMKIIδ knockouts. Unexpectedly, we found increased peribronchial eosinophils in the smooth muscle CaMKIIδ knockouts compared to control animals at 1 day post-final challenge, suggesting that lack of ASM CaMKIIδ delays the progression of AHR rather than inhibiting it. Indeed, when AHR was determined at 7 days post-final OVA challenge, CaMKIIδ knockouts showed robust AHR while AHR was fully resolved in OVA-challenged control mice. These in vivo studies demonstrate a role for smooth muscle CaMKIIδ in promoting airway inflammation and AHR and suggest a complex signaling role for CaMKIIδ in regulating ASM function. These studies confirm the diverse roles of ASM cells as immune effectors that control AHR and call for further studies into CaMKIIδ-mediated signaling in ASM cells during disease.
气道平滑肌(ASM)是变应原诱导性哮喘中的关键靶细胞,已知其与气道高反应性(AHR)和慢性气道重塑有关。虽然机制和Ca(2+)信号效应器尚未完全阐明,但已表明ASM钙稳态的变化与AHR有关。在本研究中,我们测试了ASM多功能蛋白激酶钙/钙调蛋白依赖性激酶II(CaMKII)亚型CaMKIIδ和CaMKIIγ在变应原诱导的AHR和体内气道重塑中的功能。使用特应性哮喘小鼠模型,我们证明与对照组相比,源自卵清蛋白(OVA)处理动物的ASM中CaMKIIδ蛋白上调。验证了在单独的Cre-loxp系统中条件性敲除平滑肌CaMKIIδ和CaMKIIγ的遗传方法,并使用这种功能丧失方法,在卵清蛋白(OVA)诱导的气道重塑和AHR中测试了这些CaMKII亚型的功能。在该AHR模型中,对照小鼠接受OVA处理对ASM重塑无影响,CaMKIIδ基因敲除对ASM含量无独立影响。然而,在末次OVA激发后1天,CaMKIIδ基因敲除小鼠中OVA诱导的AHR被消除。CaMKIIδ基因敲除小鼠中OVA诱导的支气管周围炎症和Th2细胞因子IL-13的支气管肺泡灌洗液(BALF)水平显著降低。出乎意料的是,我们发现与对照动物相比,在末次激发后1天,平滑肌CaMKIIδ基因敲除小鼠的支气管周围嗜酸性粒细胞增多,这表明缺乏ASM CaMKIIδ会延迟AHR的进展而不是抑制它。事实上,当在末次OVA激发后7天测定AHR时,CaMKIIδ基因敲除小鼠表现出强烈的AHR,而OVA激发的对照小鼠中的AHR已完全消退。这些体内研究证明了平滑肌CaMKIIδ在促进气道炎症和AHR中的作用,并提示CaMKIIδ在调节ASM功能中具有复杂的信号作用。这些研究证实了ASM细胞作为控制AHR的免疫效应器的多种作用,并呼吁进一步研究疾病期间ASM细胞中CaMKIIδ介导的信号传导。