Arechederra Maria, Bik Emil, Rojo Carla, Elurbide Jasmin, Elizalde María, Kruk Beata, Krasnodębski Maciej, Pertkiewicz Jan, Kozieł Sławomir, Grąt Michał, Raszeja-Wyszomirska Joanna, Rullan Maria, Alkorta-Aranburu Gorka, Oyón Daniel, Fernández-Barrena Maite G, Candels Lena S, Białek Andrzej, Krupa Łukasz, Schneider Kai M, Urman Jesús, Strnad Pavel, Trautwein Christian, Milkiewicz Piotr, Krawczyk Marcin, Ávila Matías A, Berasain Carmen
Hepatology Laboratory, Solid Tumors Program, CIMA, CCUN, University of Navarra, Pamplona, Spain.
IdiSNA, Navarra Institute for Health Research, Pamplona, Spain.
Liver Int. 2025 Apr;45(4):e70049. doi: 10.1111/liv.70049.
Primary sclerosing cholangitis (PSC) is a chronic liver disease characterised by inflammation and fibrosis of the bile ducts, conferring an increased risk of cholangiocarcinoma (CCA). However, detecting CCA early in PSC patients remains challenging due to the limited sensitivity of conventional diagnostic methods, including imaging or bile duct brush cytology during endoscopic retrograde cholangiopancreatography (ERCP). This study aims to evaluate the potential of bile cell-free DNA (cfDNA) mutational analysis, termed the Bilemut assay, as a tool for CCA detection in PSC patients.
Sixty-three PSC patients undergoing ERCP due to biliary strictures were prospectively recruited. Bile samples were collected, and cfDNA was extracted and analysed using the Oncomine Pan-Cancer Cell-Free assay. Twenty healthy liver donors were included for comparison. Samples with a mutant allele frequency (MAF) ≥ 0.1% were considered positive. Correlations between mutational status and clinical characteristics were assessed.
cfDNA mutational analysis was successful in all bile samples. Mutations predominantly in KRAS, GNAS, and TP53 were detected in 36.5% (23/63) of PSC patients, compared to 10% (2/20) of healthy donors (p = 0.0269). The clinical characteristics of Bilemut-positive and -negative patients were comparable, though there was a trend towards a lower prevalence of inflammatory bowel disease in the Bilemut-positive group. Among PSC patients diagnosed with CCA during follow-up, 75% were Bilemut-positive, suggesting an association between mutational status and malignancy risk.
Mutational analysis of cfDNA obtained from bile collected from PSC patients undergoing ERCP is feasible. Implementing the Bilemut assay may help identify patients needing closer surveillance and further imaging studies.
原发性硬化性胆管炎（PSC）是一种慢性肝病，其特征为胆管炎症和纤维化，会增加胆管癌（CCA）的发病风险。然而，由于包括内镜逆行胰胆管造影术（ERCP）期间的成像或胆管刷检细胞学等传统诊断方法的敏感性有限，在PSC患者中早期检测CCA仍然具有挑战性。本研究旨在评估胆汁游离DNA（cfDNA）突变分析（即Bilemut检测法）作为PSC患者CCA检测工具的潜力。
前瞻性招募了63例因胆道狭窄接受ERCP的PSC患者。收集胆汁样本，使用Oncomine泛癌游离细胞检测法提取并分析cfDNA。纳入20名健康肝脏供体作为对照。突变等位基因频率（MAF）≥0.1%的样本被视为阳性。评估突变状态与临床特征之间的相关性。
所有胆汁样本的cfDNA突变分析均成功。在36.5%（23/63）的PSC患者中检测到主要发生在KRAS、GNAS和TP53基因的突变，而健康供体中这一比例为10%（2/20）（p = 0.0269）。Bilemut阳性和阴性患者的临床特征具有可比性，不过Bilemut阳性组炎症性肠病的患病率有降低趋势。在随访期间被诊断为CCA的PSC患者中，75%为Bilemut阳性，这表明突变状态与恶性肿瘤风险之间存在关联。
对接受ERCP的PSC患者收集的胆汁中提取的cfDNA进行突变分析是可行的。实施Bilemut检测法可能有助于识别需要更密切监测和进一步影像学检查的患者。
