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Hypermethylation in Calca Promoter Inhibited ASC Osteogenic Differentiation in Rats with Type 2 Diabetic Mellitus.

Lei Wang1, Feng Ding1, Shaojie Shi1

  • 1State Key Laboratory of Military Stomatology & National Clinical Research Center for Oral Diseases & Shaanxi Engineering Research Center for Dental Materials and Advanced Manufacture, Department of Implantology, School of Stomatology, The Fourth Military Medical University, Xi'an, China.

Stem Cells International
|March 20, 2020
PubMed
Summary

Type 2 diabetes mellitus (T2DM) impairs stem cell bone formation via increased DNA methylation of the Calca gene. CGRP treatment and 5-azacytidine intervention restored osteogenic function in T2DM stem cells.

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Area of Science:

  • Stem cell biology
  • Epigenetics
  • Endocrinology

Background:

  • Type 2 diabetes mellitus (T2DM) environment significantly reduces osteogenic differentiation of adipose-derived stem cells (ASCs).
  • Gene sequence remains unchanged, suggesting epigenetic modifications like DNA methylation may underlie impaired osteogenesis.

Purpose of the Study:

  • Investigate DNA methylation changes in ASCs from T2DM models.
  • Identify the role of the Calca gene in T2DM-induced stem cell dysfunction.
  • Evaluate therapeutic potential of CGRP and 5-azacytidine for restoring osteogenic capacity.

Main Methods:

  • Establishment of T2DM rat models and isolation of ASCs.
  • DNA methylation sequencing and IGV analysis to identify methylation changes in the Calca gene.
  • Assessment of ASC morphology, proliferation, and osteogenic differentiation following CGRP and 5-azacytidine treatment.
  • In vivo evaluation of calvarial defect repair in T2DM rats.

Main Results:

  • Increased DNA methylation of a specific Calca gene fragment was observed in ASCs from T2DM models.
  • CGRP treatment improved proliferation and significantly enhanced osteogenic markers (ALP, calcium nodules) and gene expression in ASCs-T2DM.
  • CGRP promoted calvarial defect healing in T2DM rats in a dose-dependent manner.
  • 5-azacytidine intervention reduced Calca methylation, increasing gene expression, suggesting a role for DNMT1.

Conclusions:

  • T2DM upregulates DNA methylation in the Calca promoter region, decreasing its expression and impairing osteogenesis.
  • Calca gene product promotes osteogenic differentiation of ASCs-T2DM.
  • Targeting Calca methylation offers a potential strategy to improve osteogenic capacity in T2DM patients.