A High-Accuracy Model Based on Plasma miRNAs Diagnoses Intrahepatic Cholangiocarcinoma: A Single Center with 1001 Samples

Abstract

Objectives: Intrahepatic cholangiocarcinoma (iCCA) is a highly malignant cancer. More than 70% of patients are diagnosed at an advanced stage. The aim of this study was to evaluate the diagnostic value of plasma miR-21, miR-122, and CA19-9, hoping to establish a novel model to improve the accuracy for diagnosing iCCA.

Materials and methods: Plasma miR-21 and miR-122 were detected in 359 iCCA patients and 642 controls (healthy, benign liver lesions, other malignant liver tumors). All 1001 samples were allocated to training cohort (n = 668) and validation cohort (n = 333) in a chronological order. A logistic regression model was applied to combine these markers. Area under the receiver operating characteristic curve (AUC) was used as an accuracy index to evaluate the diagnostic performance.

Results: Plasma miR-21 and miR-122 were significantly higher in iCCA patients than those in controls. Higher plasma miR-21 level was significantly correlated with larger tumor size (p = 0.030). A three-marker model was constructed by using miR-21, miR-122 and CA19-9, which showed an AUC of 0.853 (95% CI: 0.824-0.879; sensitivity: 73.0%, specificity: 87.4%) to differentiate iCCA from controls. These results were subsequently confirmed in the validation cohort with an AUC of 0.866 (0.825-0.901). The results were similar for diagnosing early (stages 0-I) iCCA patients (AUC: 0.848) and CA19-9^negative iCCA patients (AUC: 0.795).

Conclusions: We established a novel three-marker model with a high accuracy based on a large number of participants to differentiate iCCA from controls. This model showed a great clinical value especially for the diagnosis of early iCCA and CA19-9^negative iCCA.

Keywords: CA19-9; circulating biomarker; diagnosis; intrahepatic cholangiocarcinoma; microRNA.

Figure 1

Flow chart of study design. iCCA, intrahepatic cholangiocarcinoma; HCC-iCCA, combined hepatocellular carcinoma and intrahepatic cholangiocarcinoma; ROC, receiver operating characteristics.

Figure 2

Relative expression of plasma miR-21 and miR-122 in iCCA and control patients. (A,B), The relative expression of plasma miR-21 and miR-122 in iCCA patients was significantly higher than that in control (p < 0.001). (C) The relative expression of plasma miR-21 in iCCA patients with > 5cm tumor size was significantly higher than iCCA patients with ≤ 5 cm tumor size (p = 0.009). (D,E), The relative expression of plasma miR-21 and miR-122 in iCCA patients was significantly higher than that in healthy, benign liver lesions, and other liver malignancies (all p < 0.001). **, p < 0.01; ***, p < 0.001.

Figure 3

Diagnostic performance of CA19-9, miR-21, miR-122, 2-miR model and 3-marker model for iCCA. (A,B) The demonstration of CA19-9 level and 2-miR model in entire cohort by using two-dimension scatterplot and three-dimension scatterplot. iCCA patients (87.7%, 315/359) could be diagnosed by a combination of CA19-9 level and 2-miR model (gray shadow region). Red dotted line, cut-off value for CA19-9 (34 IU/mL); blue dotted line, cut-off value for 2-miR model (−0.494). (C) Diagnostic performance of five parameters for iCCA diagnosis in the training cohort. The AUC of three-marker model was significantly larger than CA19-9 (p = 0.009), miR-21 (p < 0.001), miR-122 (p < 0.001), and 2-miR model (p < 0.001). (D,E) Diagnostic performance of three-marker model in the validation cohort and entire cohort.

Figure 4

Subgroup ROC analysis for iCCA diagnosis. (A–C) Diagnostic performance of five parameters in healthy control subgroup, benign liver lesions subgroup and other liver malignancies subgroup. Three-marker model also demonstrated a preferable diagnostic efficacy than other parameters in three subgroups (Tables S3–S5). (D,E) Diagnostic efficacy of three-marker model for iCCA with different AJCC stage. Three-marker model still possessed a good diagnostic efficacy even in early stage iCCA. (F) Diagnostic efficacy of two-miR model and three-marker model for iCCA in CA19-9^negative cohort.