Kinetics and genomic profiling of adult human and mouse β-cell maturation

Islets. Jul-Aug 2011;3(4):175-87. doi: 10.4161/isl.3.4.15881. Epub 2011 Jul 1.

Abstract

Diabetes is a multifactorial metabolic disorder defined by the loss of functional pancreatic insulin-producing β-cells. The functional maturation and dedifferentiation of adult β-cells is central to diabetes pathogenesis and to β-cell replacement therapy for the treatment of diabetes. Despite its importance, the dynamics and mechanisms of adult β-cell maturation remain poorly understood. Using a novel Pdx1/Ins1 dual fluorescent reporter lentiviral vector, we previously found that individual adult human and mouse β-cells exist in at least two differentiation states distinguishable by the activation of the rat Ins1 promoter and performed the first real-time imaging of the maturation of individual cultured β-cells. Our previous study focused on transformed (MIN6) β-cells as a model to investigatethe kinetics of β-cell maturation. In the present study, we investigated the kinetics of the maturation process in primary human and mouse β-cells and performed gene expression profiling. Gene expression profiling of FACS purified immature Pdx1 (+) /Ins1 (low) cells and mature Pdx1 (high) /Ins1 (high ) cells from cultures of human islets, mouse islets and MIN6 cells revealed that Pdx1 (+) /Ins1 (low) cells are enriched for multiple genes associated with β-cell development/progenitor cells, proliferation, apoptosis, as well as genes coding for other islet cell hormones such as glucagon. We also demonstrated that the heterogeneity in β-cell maturation states previously observed in vitro, can also be found in vivo. Collectively, these experiments contribute to the understanding of maturation, dedifferentiation and plasticity of adult pancreatic β-cells. The results have significant implications for islet regeneration and for in vitro generation of functional β-cells to treat diabetes.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adult
  • Animals
  • Cell Differentiation*
  • Cell Line, Transformed
  • Cells, Cultured
  • Gene Expression Profiling
  • Genes, Reporter
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Humans
  • Insulin / genetics
  • Insulin / metabolism
  • Insulin-Secreting Cells / physiology*
  • Insulin-Secreting Cells / ultrastructure
  • Kinetics
  • Mice
  • Mice, Inbred C57BL
  • Oligonucleotide Array Sequence Analysis
  • Promoter Regions, Genetic
  • RNA, Messenger / metabolism
  • Rats
  • Recombinant Fusion Proteins / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Single-Cell Analysis
  • Trans-Activators / genetics
  • Trans-Activators / metabolism

Substances

  • Homeodomain Proteins
  • Insulin
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Trans-Activators
  • pancreatic and duodenal homeobox 1 protein