Chesné Julie, Danger Richard, Botturi Karine, Reynaud-Gaubert Martine, Mussot Sacha, Stern Marc, Danner-Boucher Isabelle, Mornex Jean-François, Pison Christophe, Dromer Claire, Kessler Romain, Dahan Marcel, Brugière Olivier, Le Pavec Jérôme, Perros Frédéric, Humbert Marc, Gomez Carine, Brouard Sophie, Magnan Antoine
UMR_S 1087 CNRS UMR_6291, l'Institut du Thorax, Université de Nantes, CHU de Nantes, Centre National de Référence Mucoviscidose Nantes-Roscoff, Nantes, France; Université de Nantes, Nantes, France.
Université de Nantes, Nantes, France; Institut National de la Santé et de la Recherche Médicale INSERM U1064, and Institut de Transplantation Urologie Néphrologie du Centre Hospitalier Universitaire Hôtel Dieu, Nantes, France.
PLoS One. 2014 Oct 20;9(10):e109291. doi: 10.1371/journal.pone.0109291. eCollection 2014.
End-stage chronic respiratory diseases (CRD) have systemic consequences, such as weight loss and susceptibility to infection. However the mechanisms of such dysfunctions are as yet poorly explained. We hypothesized that the genes putatively involved in these mechanisms would emerge from a systematic analysis of blood mRNA profiles from pre-transplant patients with cystic fibrosis (CF), pulmonary hypertension (PAH), and chronic obstructive pulmonary disease (COPD).
Whole blood was first collected from 13 patients with PAH, 23 patients with CF, and 28 Healthy Controls (HC). Microarray results were validated by quantitative PCR on a second and independent group (7PAH, 9CF, and 11HC). Twelve pre-transplant COPD patients were added to validate the common signature shared by patients with CRD for all causes. To further clarify a role for hypoxia in the candidate gene dysregulation, peripheral blood mononuclear cells from HC were analysed for their mRNA profile under hypoxia.
Unsupervised hierarchical clustering allowed the identification of 3 gene signatures related to CRD. One was common to CF and PAH, another specific to CF, and the final one was specific to PAH. With the common signature, we validated T-Cell Factor 7 (TCF-7) and Interleukin 7 Receptor (IL-7R), two genes related to T lymphocyte activation, as being under-expressed. We showed a strong impact of the hypoxia on modulation of TCF-7 and IL-7R expression in PBMCs from HC under hypoxia or PBMCs from CRD. In addition, we identified and validated genes upregulated in PAH or CF, including Lectin Galactoside-binding Soluble 3 and Toll Like Receptor 4, respectively.
Systematic analysis of blood cell transcriptome in CRD patients identified common and specific signatures relevant to the systemic pathologies. TCF-7 and IL-7R were downregulated whatever the cause of CRD and this could play a role in the higher susceptibility to infection of these patients.
终末期慢性呼吸系统疾病（CRD）具有全身性后果，如体重减轻和易感染性。然而，此类功能障碍的机制尚未得到充分解释。我们推测，通过对患有囊性纤维化（CF）、肺动脉高压（PAH）和慢性阻塞性肺疾病（COPD）的移植前患者的血液mRNA谱进行系统分析，可能会发现与这些机制相关的基因。
首先从13例PAH患者、23例CF患者和28例健康对照（HC）中采集全血。微阵列结果在第二个独立队列（7例PAH、9例CF和11例HC）中通过定量PCR进行验证。另外纳入12例移植前COPD患者，以验证所有病因的CRD患者共有的特征。为进一步阐明缺氧在候选基因失调中的作用，对HC的外周血单个核细胞在缺氧条件下的mRNA谱进行分析。
无监督层次聚类可识别出与CRD相关的3个基因特征。一个是CF和PAH共有的，另一个是CF特有的，最后一个是PAH特有的。通过共有特征，我们验证了与T淋巴细胞活化相关的两个基因——T细胞因子7（TCF-7）和白细胞介素-7受体（IL-7R）表达下调。我们发现缺氧对HC缺氧条件下的PBMC或CRD患者的PBMC中TCF-7和IL-7R表达的调节有强烈影响。此外，我们分别鉴定并验证了PAH或CF中上调的基因，包括凝集素半乳糖结合可溶性3和Toll样受体4。
对CRD患者血细胞转录组的系统分析确定了与全身病变相关的共同特征和特异性特征。无论CRD病因如何，TCF-7 和IL-7R均下调，这可能在这些患者较高的感染易感性中起作用。
