Comincini Sergio, Manai Federico, Meazza Cristina, Pagani Sara, Martinelli Carolina, Pasqua Noemi, Pelizzo Gloria, Biggiogera Marco, Bozzola Mauro
Department of Biology and Biotechnology, University of Pavia, 27100 Pavia, Italy.
Pediatrics and Adolescentology Unit, Department of Internal Medicine and Therapeutics, University of Pavia, Fondazione IRCCS San Matteo, 27100 Pavia, Italy.
Int J Mol Sci. 2017 Feb 12;18(2):391. doi: 10.3390/ijms18020391.
Celiac disease (CD) is a severe genetic autoimmune disorder, affecting about one in 100 people, where the ingestion of gluten leads to damage in the small intestine. Diagnosing CD is quite complex and requires blood tests and intestinal biopsy examinations. Controversy exists regarding making the diagnosis without biopsy, due to the large spectrum of manifesting symptoms; furthermore, small-intestinal gastroscopy examinations have a relatively complex management in the pediatric population. To identify novel molecular markers useful to increase the sensitivity and specificity in the diagnosis of pediatric CD patients, the expression levels of two key autophagy executor genes ( and ) and their regulatory validated miRNAs (miR-17 and miR-30a, respectively) were analyzed by relative quantitative real-time-PCR on a cohort of confirmed CD patients compared to age-related controls. Among the investigated targets, the non-parametric Mann-Whitney test and ROC analysis indicated the highest significant association of with CD status in the blood, while in intestinal biopsies, all of the investigated sequences were positively associated with CD diagnosis. Nomogram-based analysis showed nearly opposite expression trends in blood compared to intestine tissue, while hierarchical clustering dendrograms enabled identifying CD and control subgroups based on specific genes and miRNA expression signatures. Next, using an established in vitro approach, through digested gliadin administration in Caco-2 cells, we also highlighted that the modulation of miR-17 endogenous levels using enriched exosomes increased the intracellular autophagosome content, thereby altering the autophagic status. Altogether, these results highlighted novel molecular markers that might be useful to increase the accuracy in CD diagnosis and in molecular-based stratification of the patients, further reinforcing the functional involvement of the regulation of the autophagy process within a digestive and autoimmune-related disorder as CD.
乳糜泻（CD）是一种严重的遗传性自身免疫性疾病，约每100人中就有1人受其影响，摄入麸质会导致小肠损伤。诊断CD相当复杂，需要进行血液检测和肠道活检检查。由于症状表现范围广泛，在不进行活检的情况下进行诊断存在争议；此外，小儿群体的小肠胃镜检查管理相对复杂。为了确定有助于提高小儿CD患者诊断敏感性和特异性的新型分子标志物，通过相对定量实时PCR分析了一组确诊的CD患者与年龄匹配的对照组中两个关键自噬执行基因（ 和 ）及其经过验证的调控性miRNA（分别为miR-17和miR-30a）的表达水平。在所研究的靶点中，非参数曼-惠特尼检验和ROC分析表明，血液中 与CD状态的关联最为显著，而在肠道活检中，所有研究序列均与CD诊断呈正相关。基于列线图的分析显示，血液与肠组织中的表达趋势几乎相反，而层次聚类树状图能够根据特定基因和miRNA表达特征识别CD和对照组亚组。接下来，我们采用一种既定的体外方法，通过在Caco-2细胞中给予消化的麦醇溶蛋白，还强调了使用富含外泌体调节miR-17内源性水平可增加细胞内自噬体含量，从而改变自噬状态。总之，这些结果突出了可能有助于提高CD诊断准确性和患者分子分层准确性的新型分子标志物，进一步强化了自噬过程调控在诸如CD等消化和自身免疫相关疾病中的功能作用。
