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METTL3 Modulates Ctsk+ Lineage Supporting Cranial Osteogenesis via Hedgehog.

R Xu1,2, R Sheng1,3, W Lin1,3

  • 1State Key Laboratory of Oral Diseases & National Center for Stomatology & National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, Sichuan, China.

Journal of Dental Research
|May 16, 2024
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Summary

Methyltransferase-like 3 (METTL3) regulates N6-methyladenosine (m6A) modification in Cathepsin K (CTSK)-positive stem cells, impacting calvarial bone development and maintenance by modulating the Hedgehog signaling pathway.

Keywords:
N6-methyladenosinebonemethylationstem cellssuturetranslational science

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Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Stem Cell Biology

Background:

  • N6-methyladenosine (m6A) modification, regulated by methyltransferase-like 3 (METTL3), is crucial for stem cell fate.
  • Calvarial bone development relies on cranial sutures and Cathepsin K (CTSK)-positive calvarial stem cells (CSCs).
  • The role of m6A in regulating CTSK+ lineage cells during calvarial development is unknown.

Purpose of the Study:

  • To investigate the function of METTL3-mediated m6A modification in CTSK+ lineage cells during calvarial bone development and maintenance.
  • To elucidate the molecular mechanisms by which METTL3 influences calvarial bone formation.

Main Methods:

  • Depletion of Mettl3 in CTSK+ lineage cells in neonatal and adult mice.
  • Analysis of calvarial bone structure, mineralization, and osteocyte morphology.
  • Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq).
  • Hedgehog (Hh) signaling pathway analysis and rescue experiments using Sufu alleles or SAG21.

Main Results:

  • Mettl3 depletion in CTSK+ cells delayed neonatal suture formation and reduced mineralization.
  • Loss of Mettl3 in adult mice impaired calvarial bone formation, increasing porosity and bone marrow cavity, and decreasing osteocyte numbers.
  • METTL3 deficiency led to reduced Hedgehog (Hh) signaling pathway activity.
  • Restoration of Hh signaling partially rescued the observed bone defects.

Conclusions:

  • METTL3 regulates CTSK+ lineage cells to support calvarial bone formation and maintenance.
  • METTL3 exerts its function by modulating the Hedgehog signaling pathway.
  • These findings offer new therapeutic strategies for skull vault osseous diseases.