Serological Assessment of Activated Fibroblasts by alpha-Smooth Muscle Actin (α-SMA): A Noninvasive Biomarker of Activated Fibroblasts in Lung Disorders.
Serological Assessment of Activated Fibroblasts by alpha-Smooth Muscle Actin (α-SMA): A Noninvasive Biomarker of Activated Fibroblasts in Lung Disorders.
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DOI：
10.1016/j.tranon.2018.11.004
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发表时间：
2019-03
影响因子：
5
通讯作者：
Nielsen MJ
中科院分区：
文献类型：
--
作者：
Holm Nielsen S;Willumsen N;Leeming DJ;Daniels SJ;Brix S;Karsdal MA;Genovese F;Nielsen MJ
目的：细胞外基质（ECM）的重塑是不同肺部疾病（如纤维化和癌症）的关键事件。结缔组织中最常见的细胞类型是成纤维细胞，激活后可转分化为肌成纤维细胞。所有的肌成纤维细胞都表达α-SMA，在肺纤维化和癌症中被发现表达上调。我们评估了α-SMA作为肺纤维化和癌症中活化成纤维细胞的无创生物标志物的潜力。方法：制备α-SMA n端单克隆抗体，建立α-SMA竞争性酶联免疫吸附法（ELISA），并对其进行技术表征。在成纤维细胞模型“罐子里的疤痕”以及特发性肺纤维化（IPF）、慢性阻塞性肺疾病（COPD）和非小细胞肺癌（NSCLC）患者的血清中测量α-SMA的水平，这些患者属于两个不同的队列。结果：建立了新的α-SMA检测方法，并通过技术验证。根据scar-in-a-jar的结果，α-SMA仅存在于TGF-β激活的成纤维细胞中。在队列1中，IPF、COPD和NSCLC患者α-SMA水平显著高于健康对照组（P = 0.04、P = 0.001和P <0.0001）。健康对照与IPF患者分离的受试者操作特征下面积（AUROC）为0.865，健康对照与COPD患者分离为0.892，健康对照与NSCLC患者分离为0.983。在队列2中，非小细胞肺癌患者α-SMA水平也显著高于健康对照组（P = 0），分离非小细胞肺癌与健康对照组的AUROC为0.715。结论：在本研究中，我们开发并验证了一种针对α-SMA n端具有竞争力的ELISA检测方法。TGF-β的加入使α-SMA水平上调，表明α-SMA在活化成纤维细胞中升高。与健康对照组相比，IPF、COPD和NSCLC患者循环中α-SMA水平显著升高。通过提供活化成纤维细胞的替代测量，该检测方法有可能作为肺部疾病的一种新型无创血清学生物标志物。
OBJECTIVES: Remodeling of the extracellular matrix (ECM) is a key event in different lung disorders, such as fibrosis and cancer. The most common cell type in the connective tissue is fibroblasts, which transdifferentiate into myofibroblasts upon activation. All myofibroblasts express α-SMA, which has been found to be upregulated in lung fibrosis and cancer. We evaluated the potential of α-SMA as a noninvasive biomarker of activated fibroblasts in lung fibrosis and cancer. METHODS: A monoclonal antibody was raised against the N-terminal of α-SMA, and a novel competitive enzyme-linked immunosorbent assay (ELISA) measuring α-SMA was developed and technically characterized. Levels of α-SMA were measured in the fibroblast model, “scar-in-a-jar”, and in serum from patients with idiopathic pulmonary fibrosis (IPF), chronic obstructive lung disorder (COPD) and non–small cell lung cancer (NSCLC) belonging to two different cohorts. RESULTS: The novel α-SMA assay was developed and validated as technically robust. Based on the scar-in-a-jar results, α-SMA was only present in the fibroblasts activated by TGF-β. In cohort 1, levels of α-SMA were significantly higher in IPF, COPD and NSCLC patients compared to healthy controls (P = 0.04, P = 0.001 and P <0.0001, respectively). The area under the receiver operating characteristics (AUROC) for separation of healthy controls from IPF patients was 0.865, healthy controls from COPD patients was 0.892 and healthy controls from NSCLC patients was 0.983. In cohort 2, levels of α-SMA were also significantly higher in NSCLC patients compared to healthy controls (P = 0) and the AUROC for separating NSCLC and healthy controls was 0.715. CONCLUSIONS: In this study we developed and validated a robust competitive ELISA assay targeting the N-terminal of α-SMA. The level of α-SMA was upregulated when adding TGF-β, indicating that α-SMA is increased in activated fibroblasts. The level of α-SMA in circulation was significantly higher in patients with IPF, COPD and NSCLC compared to healthy controls. This assay could potentially be used as a novel noninvasive serological biomarker for lung disorders by providing a surrogate measure of activated fibroblasts.
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影响因子：
5.6
作者：
Kendall RT;Feghali-Bostwick CA
通讯作者：
Feghali-Bostwick CA
影响因子：
4.2
作者：
Leeming, Diana J.;Nielsen, Mette J.;Karsdal, Morten A.
通讯作者：
Karsdal, Morten A.
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作者：
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通讯作者：
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