BACKGROUND The pathophysiology of congenital cystic adenomatoid malformations (CCAM) of the lung remains poorly understood. AIM This study aimed to identify more precisely the molecular mechanisms limited to a compartment of lung tissue, through a transcriptomic analysis of the epithelium of macrocystic forms. METHODS Tissue fragments displaying CCAM were obtained during planned surgical resections. Epithelial mRNA was obtained from cystic and normal areas after laser capture microdissection (LCM). Transcriptomic analyses were performed and the results were confirmed by RT-PCR and immunohistochemistry in independent samples. RESULTS After controlling for RNA quality, we analysed the transcriptomes of six cystic areas and five control areas. In total, 393 transcripts were differentially expressed in the epithelium, between CCAM and control areas. The most highly redundant genes involved in biological functions and signalling pathways differentially expressed between CCAM and control epithelium included TGFB2, TGFBR1, and MAP 2 K1. These genes were considered particularly relevant as they have been implicated in branching morphogenesis. RT-qPCR analysis confirmed in independent samples that TGFBR1 was more strongly expressed in CCAM than in control tissues (p < 0.03). Immunohistochemistry analysis showed TGFBR1 (p = 0.0007) and TGFB2 (p < 0.02) levels to be significantly higher in the epithelium of CCAM than in that of control tissues. CONCLUSIONS This compartmentalised transcriptomic analysis of the epithelium of macrocystic lung malformations identified a dysregulation of TGFB signalling at the mRNA and protein levels, suggesting a possible role of this pathway in CCAM pathogenesis. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT01732185.

中文翻译：

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先天性肺囊性腺瘤样畸形：上皮转录组学方法。

背景技术对肺的先天性囊性腺瘤样畸形（CCAM）的病理生理学知之甚少。目的本研究旨在通过对大囊形式的上皮进行转录组分析，以更精确地确定局限于肺组织隔室的分子机制。方法在计划的手术切除过程中获得显示CCAM的组织碎片。激光捕获显微切割（LCM）后，从囊性和正常区域获得上皮mRNA。进行了转录组学分析，结果通过独立样本中的RT-PCR和免疫组织化学证实。结果在控制了RNA质量之后，我们分析了六个囊性区域和五个对照区域的转录组。总共有393个转录本在CCAM和对照区之间的上皮中差异表达。在CCAM和对照上皮之间差异表达的生物学功能和信号通路中涉及的最高冗余基因包括TGFB2，TGFBR1和MAP 2 K1。这些基因被认为特别相关，因为它们与分支形态发生有关。RT-qPCR分析在独立样品中证实，TGFBR1在CCAM中的表达比在对照组织中更强（p <0.03）。免疫组织化学分析显示，CCAM上皮中的TGFBR1（p = 0.0007）和TGFB2（p <0.02）水平显着高于对照组织。结论这项对大囊性肺畸形上皮的间隔转录组学分析确定了TGFB信号在mRNA和蛋白水平上的失调，提示此途径在CCAM发病机理中的可能作用。试验注册ClinicalTrials.gov标识符：NCT01732185。