MicroRNA Sequencing Identifies a Serum MicroRNA Panel, Which Combined With Aspartate Aminotransferase to Platelet Ratio Index Can Detect and Monitor Liver Disease in Pediatric Cystic Fibrosis

Abstract

Cystic fibrosis (CF)-associated liver disease (CFLD) is a hepatobiliary complication of CF. Current diagnostic modalities rely on nonspecific assessments, whereas liver biopsy is the gold standard to assess severity of fibrosis. MicroRNAs (miRNAs) regulate liver disease pathogenesis and are proposed as diagnostic biomarkers. We investigated the combined use of serum miRNAs and aspartate aminotransferase (AST) to platelet ratio (APRI) to diagnose and assess CFLD severity. This was a cross-sectional cohort study of the circulatory miRNA signature of 124 children grouped by clinical, biochemical, and imaging assessments as follows: CFLD (n = 44), CF patients with no evidence of liver disease (CFnoLD; n = 40), and healthy controls (n = 40). Serum miRNAs were analyzed using miRNA sequencing (miRNA-Seq). Selected differentially expressed serum miRNA candidates were further validated by qRT-PCR and statistical analysis performed to evaluate utility to predict CFLD and fibrosis severity validated by liver biopsy, alone or in combination with APRI. Serum miR-122-5p, miR-365a-3p, and miR-34a-5p levels were elevated in CFLD compared to CFnoLD, whereas miR-142-3p and let-7g-5p were down-regulated in CFLD compared to CFnoLD. Logistic regression analysis combining miR-365a-3p, miR-142-3p, and let-7g-5p with APRI showed 21 times greater odds of accurately predicting liver disease in CF with an area under the receiver operating characteristics curve (AUROC) = 0.91 (sensitivity = 83%, specificity = 92%; P < 0.0001). Expression levels of serum miR-18a-5p were correlated with increasing hepatic fibrosis (HF) stage in CFLD (r = 0.56; P < 0.0001), showing good diagnostic accuracy for distinguishing severe (F3-F4) from mild/moderate fibrosis (F0-F2). A unit increase of miR-18a-5p showed a 7-fold increased odds of having severe fibrosis with an AUROC = 0.82 (sensitivity = 93%, specificity = 73%; P = 0.004), indicating its potential to predict fibrosis severity. Conclusion: We identified a distinct circulatory miRNA profile in pediatric CFLD with potential to accurately discriminate liver disease and fibrosis severity in children with CF.

miR-455-3p Alleviates Hepatic Stellate Cell Activation and Liver Fibrosis by Suppressing HSF1 Expression

“…In recent years, researchers have focused on the role of miRNAs in the pathophysiology of hepatic fibrosis to determine its role in the regulation of HSC proliferation and differentiation 17 . Aberrant expression of several miRNAs was identified between resting and activated HSCs 18 . A recent study shows that miR-455-3p served as oncogene or anti-oncogene in different types of tumors 19 .…”

Accuracy of Transient Elastography Data Combined With APRI in Detection and Staging of Liver Disease in Pediatric Patients With Cystic Fibrosis

“…Supplementary Figure 1D) or CFLD (r=-0.1, p=0.6, not shown). We previously demonstrated the use of APRI for detection of liver disease and in staging fibrosis severity in pediatric CFLD 15,20 . In the current cohort, contemporaneous blood test results were available for calculation of APRI in 72/105 subjects with CFnoLD and 23/33 with CFLD.…”

“…Different non-invasive assessment tools have been assessed to improve diagnostic accuracy for both liver disease detection in pediatric CF and hepatic fibrosis staging in CFLD including APRI, Fibrosis-4 (FIB-4) and serum microRNAs 15,20 . In this study, by combining LSM and APRI in a CART recursive partition model, the AUROC for detecting CFLD improved to 0.89, and predicted 18.6-times greater odds of children with CF having CFLD.…”

Non-invasive assessment of paediatric cystic fibrosis liver disease and the role of microRNAs in disease mechanism

“…(D) Scatterplot illustrates cut-points determined by CART modelling used to discriminate CFLD from CFnoLD children. Blue dots represent CFnoLD.Red dots represent CFLDSeveral studies have shown the use of APRI as a non-invasive biomarker for the detection of CFLD(124,232). ROC curve analysis of APRI for detection of liver disease in this cohort (i.e., CFLD vs CFnoLD) showed an AUC= 0.74 (P=0.0040), similar to previous reports(124).…”