Evaluation of circulating cell-free DNA in cholestatic liver disease using liver-specific methylation markers.
Evaluation of circulating cell-free DNA in cholestatic liver disease using liver-specific methylation markers.
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DOI：
10.1186/s12876-021-01741-5
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发表时间：
2021-04-01
影响因子：
2.4
通讯作者：
Lazaridis KN
中科院分区：
文献类型：
--
作者：
Punia S;Juran BD;Ali AH;Schlicht EM;Moore RM;Sun Z;Lazaridis KN
循环中器官特异性无细胞DNA(CfDNA)的定量提供了一种正在进行的细胞死亡的敏感指标，有助于评估缺乏可靠非侵入性生物标志物的胆汁淤积性肝病原发性胆管炎(PBC)和原发性硬化性胆管炎(PSC)。在这项先导性研究中，我们的目标是确定PBC和PSC患者的肝脏特异性cfDNA水平是否比对照组和晚期疾病与早期疾病患者高，并评估它们作为新的疾病生物标志物的潜力。用聚合酶链式反应扩增肝癌患者(n = 48例)、原发性肝癌(n = 48例)和对照组(n = 96例)外周血中亚硫酸氢盐处理过的DNA，以评估肝组织IGF2R、ITIH4和Vtn基因附近16个CpG位点的甲基化状态。用扩增片段制备成对的末端文库，在MiSeq测序仪上测序。对修剪后的读数进行比对，并用于确定去甲基化比率和计算肝脏特异性cfDNA浓度。两组之间的比较使用双尾Mann-Whitney检验，变量之间的关系使用皮尔逊相关性进行评估。PBC和PSC患者的肝脏特异性cfDNA水平在3个基因位点上的测量结果均高于对照组，晚期患者的水平高于早期患者。此外，在患者中，cfDNA水平与碱性磷酸酶水平相关，碱性磷酸酶是一种常用的生化测试，用于评估肝病的疾病严重程度，但在对照组中没有相关性。CfDNA作为一种无创的液体活检方法，可用于评估胆汁淤积性肝病患者的肝脏特异性细胞死亡。
Quantification of circulating organ-specific cell-free DNA (cfDNA) provides a sensitive measure of ongoing cell death that could benefit evaluation of the cholestatic liver diseases primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC), which lack reliable non-invasive biomarkers. Our goal in this pilot study was to determine whether liver-specific cfDNA levels are increased in PBC and PSC patients relative to controls and in advanced versus early disease, to evaluate their potential as novel disease biomarkers. Peripheral blood derived bisulfite-treated DNA was PCR amplified from patients with PBC (n = 48), PSC (n = 48) and controls (n = 96) to evaluate methylation status at 16 CpG sites reported to be specifically unmethylated in liver tissue near the genes IGF2R, ITIH4 and VTN. Amplicons were used to prepare paired end libraries which were sequenced on a MiSeq sequencer. Trimmed reads were aligned and used to determine unmethylation ratios and to calculate concentration of liver-specific cfDNA. Comparisons between groups were performed using the two-tailed Mann–Whitney Test and relationships between variables were evaluated using Pearson’s Correlation. Levels of liver-specific cfDNA, as measured at the 3 genetic loci, were increased in PBC and PSC patients relative to controls and in late-stage relative to early-stage patients. As well, cfDNA levels were correlated with levels of alkaline phosphatase, a commonly used biochemical test to evaluate disease severity in liver disease, in patients, but not in controls. cfDNA offers promise as a non-invasive liquid-biopsy to evaluate liver-specific cell-death in patients with cholestatic liver diseases.
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影响因子：
24.5
作者：
Hardy T;Zeybel M;Day CP;Dipper C;Masson S;McPherson S;Henderson E;Tiniakos D;White S;French J;Mann DA;Anstee QM;Mann J
通讯作者：
Mann J
DOI：
10.1056/nejmra1506330
发表时间：
2016-09-22
期刊：
The New England journal of medicine
影响因子：
--
作者：
Lazaridis KN;LaRusso NF
通讯作者：
LaRusso NF
严重败血症患者的预后效用和无细胞DNA的表征。
DOI：
10.1186/cc11466
发表时间：
2012-08-13
期刊：
Critical care (London, England)
影响因子：
--
作者：
Dwivedi DJ;Toltl LJ;Swystun LL;Pogue J;Liaw KL;Weitz JI;Cook DJ;Fox-Robichaud AE;Liaw PC;Canadian Critical Care Translational Biology Group
通讯作者：
Canadian Critical Care Translational Biology Group
影响因子：
6.3
作者：
Bakhshi Z;Hilscher MB;Gores GJ;Harmsen WS;Viehman JK;LaRusso NF;Gossard AA;Lazaridis KN;Lindor KD;Eaton JE
通讯作者：
Eaton JE
影响因子：
8
作者：
Cai, Shi-Ying;Ouyang, Xinshou;Boyer, James L.
通讯作者：
Boyer, James L.