RNA-Seq Analysis of Islets to Characterise the Dedifferentiation in Type 2 Diabetes Model Mice db/db

Endocr Pathol. 2018 Sep;29(3):207-221. doi: 10.1007/s12022-018-9523-x.

Abstract

Type 2 diabetes (T2D) is a global health issue and dedifferentiation plays underlying causes in the pathophysiology of T2D; however, there is a lack of understanding in the mechanism. Dedifferentiation results from the loss of function of pancreatic β-cells alongside a reduction in essential transcription factors under various physiological stressors. Our study aimed to establish db/db as an animal model for dedifferentiation by using RNA sequencing to compare the gene expression profile in islets isolated from wild-type, db/+ and db/db mice, and qPCR was performed to validate those significant genes. A reduction in both insulin secretion and the expression of Ins1, Ins2, Glut2, Pdx1 and MafA was indicative of dedifferentiation in db/db islets. A comparison of the db/+ and the wild-type islets indicated a reduction in insulin secretion perhaps related to the decreased Mt1. A significant reduction in both Rn45s and Mir6236 was identified in db/+ compared to wild-type islets, which may be indicative of pre-diabetic state. A further significant reduction in RasGRF1, Igf1R and Htt was also identified in dedifferentiated db/db islets. Molecular characterisation of the db/db islets was performed via Ingenuity analysis which identified highly significant genes that may represent new molecular markers of dedifferentiation.

Keywords: Db/db islets; Network analysis; β-cell dedifferentiation.

MeSH terms

  • Animals
  • Cell Dedifferentiation / genetics*
  • Diabetes Mellitus, Experimental
  • Diabetes Mellitus, Type 2 / genetics*
  • Female
  • Gene Expression Profiling
  • Islets of Langerhans*
  • Male
  • Mice
  • Sequence Analysis, RNA
  • Transcriptome