Serum cell-free DNA methylation of OPCML and HOXD9 as a biomarker that may aid in differential diagnosis between cholangiocarcinoma and other biliary diseases

Abstract

Background Cholangiocarcinoma (CCA) is a fatal cancer of the bile duct epithelial cell lining. The misdiagnosis of CCA and other biliary diseases may occur due to the similarity of clinical manifestations and blood tests resulting in inappropriate or delayed treatment. Thus, an accurate and less-invasive method for differentiating CCA from other biliary diseases is inevitable. Methods We quantified methylation of OPCML , HOXA9 , and HOXD9 in serum cell-free DNA (cfDNA) of CCA patients and other biliary diseases using methylation-sensitive high-resolution melting (MS-HRM). Their potency as differential biomarkers between CCA and other biliary diseases was also evaluated by using receiver operating characteristic (ROC) curves. Results The significant difference of methylation levels of OPCML and HOXD9 was observed in serum cfDNA of CCA compared to other biliary diseases. Assessment of serum cfDNA methylation of OPCML and HOXD9 as differential biomarkers of CCA and other biliary diseases showed the area under curve (AUC) of 0.850 (0.759–0.941) for OPCML which sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were 80.00%, 90.00%, 88.88%, 81.81%, and 85.00%, respectively. The AUC of HOXD9 was 0.789 (0.686–0.892) with sensitivity, specificity, PPV, NPV, and accuracy of 67.50%, 90.00%, 87.09%, 73.46%, and 78.75%, respectively. The combined marker between OPCML and HOXD9 showed sensitivity, specificity, PPV, and NPV of 62.50%, 100%, 100%, and 72.72%, respectively, which may be helpful to prevent a misdiagnosis between CCA and other biliary diseases. Conclusions Our findings suggest the application of serum cfDNA methylation of OPCML and HOXD9 for differential diagnosis of CCA and other biliary diseases due to its less invasiveness and clinically practical method which may benefit the patients by preventing the misdiagnosis of CCA and avoiding unnecessary surgical intervention. Electronic supplementary material The online version of this article (10.1186/s13148-019-0634-0) contains supplementary material, which is available to authorized users.

Methylation in HOX Clusters and Its Applications in Cancer Therapy

“…Biomarkers for the detection of cholangiocarcinoma in tissues or serum cell-free. [31][32][33] Bladder HOXA9…”

Epigenetic Biomarkers in the Management of Ovarian Cancer: Current Prospectives

“…Similarly, promoter methylation of OPCML and HOXD9 assessed in serum cell free DNA using methylation-sensitive high-resolution melting, was detected with a sensitivity of 62.50% with specificity of 100%, thus could serve as a noninvasive differential biomarker to prevent misdiagnosis of cholangiocarcinoma (CCA) and other biliary diseases (Wasenang et al, 2019). Further, for the management of pancreatic cancer and its early detection Eissa et al analyzed the methylation of ADAMTS1 and BNC1 in cfDNA using qMSP, which exhibited a sensitivity of 94.8% and specificity of 91.6% with a AUC of 0.95 reflecting diagnostic potential of this blood based two-gene panel in detection of pancreatic cancer at an early stage (Eissa et al, 2019).…”

Detecting cholangiocarcinoma in patients with primary sclerosing cholangitis – The promise of DNA methylation and molecular biomarkers

“…43 OPCML methylation was originally identified in tissue samples, 34 and was also recently found in serum from 80% of CCA samples (n = 40) and in only 10% of samples from people with benign biliary disorders (90% specificity). 44 If methylation of OPCML was combined with methylation of HOXD9 the specificity increased to 100%, but the sensitivity decreased to 63%. 44 No blood samples from PSC controls were analysed in either of these studies.…”

“…44 If methylation of OPCML was combined with methylation of HOXD9 the specificity increased to 100%, but the sensitivity decreased to 63%. 44 No blood samples from PSC controls were analysed in either of these studies. 43,44 Molecular biomarkers for detection of CCA in patients with PSC In addition to DNA methylation markers, several other biomarkers, including microRNAs (miRNAs) and peptide/protein markers have been suggested as potential candidates for improved detection of CCA among patients with PSC (Table 2).…”

“…44 No blood samples from PSC controls were analysed in either of these studies. 43,44 Molecular biomarkers for detection of CCA in patients with PSC In addition to DNA methylation markers, several other biomarkers, including microRNAs (miRNAs) and peptide/protein markers have been suggested as potential candidates for improved detection of CCA among patients with PSC (Table 2). In the current review, only markers detected in liquid biopsies (blood, bile and urine) or biliary brushes have been included.…”