細胞外小胞が歯周組織の再生を調節する仕組みを解明ーmicroRNAによる幹細胞の運命制御を明らかに 歯周炎の新たな無細胞治療へー

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2026-09-28 東京科学大学

東京科学大学の研究グループは、ヒト歯根膜由来間葉系間質細胞(hPDL-MSC)が分泌する細胞外小胞(EV)に含まれるmicroRNAが、歯周組織再生に関わる幹細胞の運命を制御する仕組みを解明した。特にmiR-181c-5pはRNF150を標的としてMAP3K5-p38経路を活性化し、骨芽細胞への分化を促進する一方、let-7c-3pはNT5E/CD73を介して細胞増殖や幹細胞性を促進し、骨芽細胞分化への移行を抑制することを明らかにした。さらに、マウス歯周炎モデルへのhPDL-MSC由来EVの局所投与により、炎症に伴う歯槽骨破壊の抑制を確認した。EV中の機能性miRNAを調整することで、細胞そのものを移植しない歯周組織の無細胞再生治療につながる可能性が示された。

細胞外小胞が歯周組織の再生を調節する仕組みを解明ーmicroRNAによる幹細胞の運命制御を明らかに 歯周炎の新たな無細胞治療へー
図1. 研究デザインとhPDL-MSC由来EVの性状・機能評価。
(A)研究デザインの模式図。(B)異なる継代数(P7、P10)および温度条件(37℃、25℃)で得たEVを処理したOsteo_High/Osteo_Low細胞由来EVを投与した際のアリザリンレッドS染色。

<関連情報>

細胞外小胞からの標的遺伝子miR-181cシグナル伝達は、RNF150-MAP3K5を介して歯周MSCの運命を制御する Targetome-defined miR-181c signaling from extracellular vesicles governs periodontal MSC fate via RNF150-MAP3K5

Jingyi Gao, Anhao Liu, Ye Yint Kaung Myint, Masahiro Hatasa, Supreda Suphanantachat Srithanyarat & Takanori Iwata
Cell Communication and Signaling  Published:23 September 2026
DOI:https://doi.org/10.1186/s12964-026-03153-y

Abstract

Background
In regenerative settings, robust differentiation of human mesenchymal stromal cells (hMSCs) requires precise coupling between post-transcriptional regulation and kinase-driven pathways to achieve optimal therapeutic efficacy. miRNA-mediated regulation is widely implicated in stem cell fate control, yet its mechanistic contribution to lineage commitment remains incompletely defined.

Methods
Multi-conditioned periodontal ligament–derived hMSCs were developed as the regenerative model for alveolar bone. Extracellular vesicle (EV)-encapsulated miRNAs were delineated through microarrays. AGO2 RNA-immunoprecipitation sequencing with transcriptome profiling was integrated to establish the miRISC-associated targetome. Protein-protein interactions and signaling hierarchy were examined by Co-IP, WB, and IFC under knockdown/overexpression with pharmacological interventions. The translational efficacy of EV and miRNA was validated using a mouse ligature-induced periodontitis model, followed by µCT and histological analyses.

Results
EVs produced by hMSCs with higher intrinsic osteogenic capacity exhibited enhanced osteo-inductivity, traced to the enrichment of miR-181c. miR-181c was consistently upregulated during osteogenesis, with gain- and loss-of-function producing concordant effects. Mechanistically, the E3 ubiquitin ligase RNF150 emerged as the main target, whose repression reduced MAP3K5 ubiquitin-mediated proteolysis and enabled p38 activation. Local administration of EVs and miR-181c both promoted alveolar bone regeneration process in vivo.

Conclusion
Collectively, EV–miR-181c–RNF150–MAP3K5–p38 axis was proposed linking miRNA-mediated repression to kinase activation and lineage commitment. This work provides the framework for how EV-delivered miRNAs gate hMSCs’ fate decision, with significance for acellular therapeutic modalities in periodontal and skeletal tissues.

 

hsa-let-7c-3pはNT5Eを介してヒト歯周靭帯間葉系幹細胞/間質細胞の増殖から骨形成への移行を抑制する hsa-let-7c-3p Restrains the Proliferation-to-Osteogenesis Transition of Human Periodontal Ligament Mesenchymal Stem/Stromal Cells via NT5E

Ye Yint Kaung Myint, Anhao Liu, Jingyi Gao, Masahiro Hatasa, Supreda Suphanantachat Srithanyarat & Takanori Iwata
Stem Cell Reviews and Reports  Published:02 June 2026
DOI:https://doi.org/10.1007/s12015-026-11160-2

Abstract

Small extracellular vesicles (sEVs) derived from mesenchymal stem/stromal cells (MSCs) contain microRNAs (miRNAs) that can modulate the cellular behavior of recipient cells. Human periodontal ligament mesenchymal stem/stromal cells (hPDL-MSCs) are a unique MSC population involved in periodontal tissue regeneration; however, the specific miRNAs that regulate their proliferative and differentiation–related properties remain incompletely understood. In this study, we profiled miRNAs enriched in sEVs secreted by highly proliferative human PDL-MSCs and identified hsa-let-7c-3p as a candidate regulator of hPDL-MSC behavior. Functional assays demonstrated that hsa-let-7c-3p enhanced proliferative and migratory behavior, reflected by an approximately 1.5-fold increase in metabolic activity and a 2-fold increase in migration, while suppressing osteogenic differentiation, with approximately 50% reductions in alkaline phosphatase activity and mineralization. To identify functionally relevant downstream targets, we intersected AGO2-RIP-seq–enriched transcripts, RNA-seq–defined genes altered by hsa-let-7c-3p overexpression, and in silico target predictions, identifying NT5E/CD73 as a candidate downstream target with experimental validation. Furthermore, NT5E knockdown partially phenocopied the effects of hsa-let-7c-3p overexpression on hPDL-MSC proliferation, migration, and osteogenic differentiation, supporting a functional link between hsa-let-7c-3p and NT5E/CD73. These findings highlight hsa-let-7c-3p as an underexplored sEV-associated 3p miRNA strand that restrains the proliferation-to-osteogenesis transition of hPDL-MSCs and identify NT5E/CD73 as a functionally relevant downstream target. Clinical trial number. Not applicable.

細胞遺伝子工学
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