Differential regulation of serum microRNA expression by HNF1β and HNF1α transcription factors

Diabetologia. 2016 Jul;59(7):1463-1473. doi: 10.1007/s00125-016-3945-0. Epub 2016 Apr 8.

Abstract

Aims/hypothesis: We aimed to identify microRNAs (miRNAs) under transcriptional control of the HNF1β transcription factor, and investigate whether its effect manifests in serum.

Methods: The Polish cohort (N = 60) consisted of 11 patients with HNF1B-MODY, 17 with HNF1A-MODY, 13 with GCK-MODY, an HbA1c-matched type 1 diabetic group (n = 9) and ten healthy controls. Replication was performed in 61 clinically-matched British patients mirroring the groups in the Polish cohort. The Polish cohort underwent miRNA serum level profiling with quantitative real-time PCR (qPCR) arrays to identify differentially expressed miRNAs. Validation was performed using qPCR. To determine whether serum content reflects alterations at a cellular level, we quantified miRNA levels in a human hepatocyte cell line (HepG2) with small interfering RNA knockdowns of HNF1α or HNF1β.

Results: Significant differences (adjusted p < 0.05) were noted for 11 miRNAs. Five of them differed between HNF1A-MODY and HNF1B-MODY, and, amongst those, four (miR-24, miR-27b, miR-223 and miR-199a) showed HNF1B-MODY-specific expression levels in the replication group. In all four cases the miRNA expression level was lower in HNF1B-MODY than in all other tested groups. Areas under the receiver operating characteristic curves ranged from 0.79 to 0.86, with sensitivity and specificity reaching 91.7% (miR-24) and 82.1% (miR-199a), respectively. The cellular expression pattern of miRNA was consistent with serum levels, as all were significantly higher in HNF1α- than in HNF1β-deficient HepG2 cells.

Conclusions/interpretation: We have shown that expression of specific miRNAs depends on HNF1β function. The impact of HNF1β deficiency was evidenced at serum level, making HNF1β-dependent miRNAs potentially applicable in the diagnosis of HNF1B-MODY.

Keywords: HNF; MODY; Monogenic diabetes; Transcription factors; microRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Expression Regulation / genetics
  • Gene Expression Regulation / physiology
  • Glycated Hemoglobin A / genetics
  • Glycated Hemoglobin A / metabolism
  • Hep G2 Cells
  • Hepatocyte Nuclear Factor 1-alpha / genetics
  • Hepatocyte Nuclear Factor 1-alpha / metabolism*
  • Hepatocyte Nuclear Factor 1-beta / genetics
  • Hepatocyte Nuclear Factor 1-beta / metabolism*
  • Humans
  • MicroRNAs / blood*
  • MicroRNAs / genetics*
  • RNA, Small Interfering / genetics
  • ROC Curve
  • Real-Time Polymerase Chain Reaction

Substances

  • Glycated Hemoglobin A
  • Hepatocyte Nuclear Factor 1-alpha
  • MicroRNAs
  • RNA, Small Interfering
  • hemoglobin A1c protein, human
  • Hepatocyte Nuclear Factor 1-beta