Dual-parameter liquid biopsy using plasma miR-21-5p and T cell LAIR-1 mean fluorescence intensity for hepatocellular carcinoma diagnosis in a high-risk Egyptian cohort

Abstract

BACKGROUND

The early detection of hepatocellular carcinoma (HCC) in patients with liver cirrhosis remains a major clinical challenge, particularly in Egypt, with its high burden of hepatitis C virus-related disease. Serum α-fetoprotein lacks sufficient sensitivity, creating a need for robust noninvasive biomarkers.

AIM

To evaluate a dual-parameter liquid biopsy approach by comparing a tumor-derived circulating microRNA and an immune-related marker for discriminating HCC from cirrhosis.

METHODS

A prospective observational case-control study included 211 participants: 70 with HCC; 60 with hepatitis C virus-related liver cirrhosis; and 81 healthy controls. Plasma microRNAs were quantified by quantitative real-time PCR using SNORD68 as the endogenous control. Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) expression on T cells was measured by flow cytometry and expressed as percentage of LAIR-1⁺ T cells and mean fluorescence intensity (MFI). Data were analyzed using non-parametric tests (Kruskal-Wallis with Dunn’s post hoc), and diagnostic performance was assessed by receiver operating characteristic curves, area under the curve (AUC), and Youden index-derived sensitivity and specificity. Statistical significance was set at P < 0.05.

RESULTS

Plasma levels of hsa-miR-21-5p, hsa-miR-155-5p, and LAIR-1 expression were significantly elevated in the HCC group compared with the cirrhosis and control groups (P < 0.001). For distinguishing HCC from cirrhosis, both hsa-miR-21-5p and LAIR-1 MFI demonstrated outstanding diagnostic performance with AUCs of 0.990 [95% confidence interval: 0.974-1.000] and 0.997 (95% confidence interval: 0.990-1.000), respectively. At optimal cutoffs hsa-miR-21-5p showed a sensitivity of 98.6% and specificity of 96.7% while LAIR-1 MFI showed 100% sensitivity and 98.3% specificity. hsa-miR-155-5p and the percentage of LAIR-1⁺ T cells showed lower but significant performance (AUC = 0.874 and 0.833, respectively). Both leading biomarkers maintained exceptional accuracy (AUC > 0.98) in critical subgroups, including early-stage and α-fetoprotein-negative HCC. In a multivariable model, which showed no significant multicollinearity (all variance inflation factor < 5), LAIR-1 MFI emerged as the sole independent predictor of HCC (odds ratio = 1.484; P = 0.028).

CONCLUSION

This dual-parameter approach, integrating plasma hsa-miR-21-5p and T cell LAIR-1 MFI, achieved exceptional diagnostic accuracy for HCC in a high-risk cohort. While their individual performance was comparable, LAIR-1 MFI provides unique, independent diagnostic information. These findings nominate a promising, integrated liquid biopsy strategy to improve HCC surveillance.

Keywords: Hepatocellular carcinoma; MicroRNAs; hsa-miR-21-5p; hsa-miR-155-5p; Leukocyte-associated immunoglobulin-like receptor-1; Biomarkers; Receiver operating characteristic curve; Egypt

Core Tip: Early, accurate, and noninvasive diagnosis of hepatocellular carcinoma (HCC) remains a major clinical challenge, particularly in Egypt, where hepatitis C virus-related cirrhosis is prevalent. This study introduced a novel dual-parameter liquid biopsy combining plasma miR-21-5p (a tumor-derived oncogenic microRNA) and T cell leukocyte-associated immunoglobulin-like receptor-1 mean fluorescence intensity (an immune checkpoint marker). In a prospective Egyptian cohort, this approach achieved exceptional diagnostic accuracy (area under the curve > 0.99) and maintained high sensitivity in early-stage and α-fetoprotein-negative HCC. The findings proposed an integrated molecular-immune diagnostic pathway that could substantially improve HCC surveillance and early detection in high-risk populations.