Early and accurate detection of cholangiocarcinoma in patients with primary sclerosing cholangitis by methylation markers in bile

Abstract

Background and Aims: Primary sclerosing cholangitis (PSC) is associated with increased risk of cholangiocarcinoma (CCA). Early and accurate CCA detection represents an unmet clinical need as the majority of patients with PSC are diagnosed at an advanced stage of malignancy. In the present study, we aimed at establishing robust DNA methylation biomarkers in bile for early and accurate diagnosis of CCA in PSC. Approach and Results: Droplet digital PCR (ddPCR) was used to analyze 344 bile samples from 273 patients with sporadic and PSC-associated CCA, PSC, and other nonmalignant liver diseases for promoter methylation of cysteine dioxygenase type 1, cannabinoid receptor interacting protein 1, septin 9, and vimentin. Receiver operating characteristic (ROC) curve analyses revealed high AUCs for all four markers (0.77-0.87) for CCA detection among patients with PSC. Including only samples from patients with PSCdiagnosed with CCA ≤ 12 months following bile collection increased the accuracy for cancer detection, with a combined sensitivity of 100% (28/28) and a specificity of 90% (20/203). The specificity increased to 93% when only including patients with PSC with longtime follow-up (> 36 months) as controls, and remained high (83%) when only including patients with PSC and dysplasia as controls (n = 23). Importantly, the bile samples from the CCA-PSC

Clinical relevance of biomarkers in cholangiocarcinoma: critical revision and future directions

“…Bile is also emerging as a promising liquid biopsy matrix for CCA diagnosis; a recent study showed that detection of mutations in bile cell-free DNA has a strong sensitivity for early malignancy detection within the biliary tract 40. Identification of differentially methylated markers in bile cell-free DNA has also been recently demonstrated to provide high sensitivity for CCA detection 41 42. If confirmed, these approaches may significantly improve the management of patients with biliary strictures in which intraductal biopsy results inconclusive and further support the biliary fluid as a relevant matrix for relatively low invasive liquid biopsy.…”

Diagnosis Biomarkers of Cholangiocarcinoma in Human Bile: An Evidence-Based Study

“…Droplet digital PCR (ddPCR) can absolutely quantify DNA methylation in samples. As absolute quantification of DNA methylation in samples was possible by ddPCR; Vedeld et al analyzed 344 bile samples containing CCA and other benign biliary diseases by ddPCR, indicating the sensitivity and specificity of combing diagnosis of CDO1, CNRIP1, SEPT9 and VIM promoter methylation were 100% and 90%, respectively [ 10 ]. The earliest incidence of tumor growth is generally genetic variation, and methylight technology is reasonably developed and provides advantages in the detection of early CCA.…”

“…For different types of bile candidate markers, treatment and analysis methods are as follows: (1) Gene level: After extracting DNA from the bile, amplification of the target fragment by PCR to obtain the expression of potential diagnostic biomarkers and analysis of data [ 8 ]. For RNA, target RNA reverse transcription is required to form a stable CDNA before PCR amplification and data analysis, and real-time quantitative PCR is usually used [ 9 , 10 ]. (2) Protein level: enzyme linked immunosorbent assay (ELISA) or automatic immunofluorescence analysis is adopted to verify suspicious protein biomarkers in bile, but when exploring unknown diagnostic biomarkers, proteomics methods are being generally used to find markers and then checked by ELISA in bile [ 11 , 12 , 13 ].…”

Silicon Nanowires Length and Numbers Dependence on Sensitivity of the Field-Effect Transistor Sensor for Hepatitis B Virus Surface Antigen Detection

“…Because the liver does not have nerves, those with HBV infection do not notice any symptoms in the initial stages. By the time the body begins to sends warning signals, the optimal treatment period has already passed [ 31 , 32 ]. Although medication can be used to treat hepatitis B, the recovery rate is low.…”