PMID- 37162558 OWN - NLM STAT- MEDLINE DCOM- 20230629 LR - 20231213 IS - 1432-1440 (Electronic) IS - 0946-2716 (Linking) VI - 101 IP - 7 DP - 2023 Jul TI - LAMP2A regulates the balance of mesenchymal stem cell adipo-osteogenesis via the Wnt/beta-catenin/GSK3beta signaling pathway. PG - 783-799 LID - 10.1007/s00109-023-02328-1 [doi] AB - Chaperone-mediated autophagy (CMA) plays multiple roles in cell metabolism. We found that lysosome-associated membrane protein type 2A (LAMP2A), a crucial protein of CMA, plays a key role in the control of mesenchymal stem cell (MSC) adipo-osteogenesis. We identified a differentially expressed CMA gene (LAMP2) in GEO datasets (GSE4911 and GSE494). Further, we performed co-expression analyses to define the relationships between CMA components genes and other relevant genes including Col1a1, Runx2, Wnt3 and Gsk3beta. Mouse BMSCs (mMSCs) exhibiting Lamp2a gene knockdown (LA-KD) and overexpression (LA-OE) were created using an adenovirus system; then we investigated LAMP2A function in vitro by Western blot, Oil Red staining, ALP staining, ARS staining and Immunofluorescence analysis. Next, we used a modified mouse model of tibial fracture to investigate LAMP2A function in vivo. LAMP2A knockdown in mMSCs decreased the levels of osteogenic-specific proteins (COL1A1 and RUNX2) and increased those of the adipogenesis markers PPARgamma and C/EBPalpha; LAMP2A overexpression had the opposite effects. The active-beta-catenin and phospho-GSK3beta (Ser9) levels were upregulated by LAMP2A overexpression and downregulated by LAMP2A knockdown. In the mouse model of tibial fracture, mMSC-overexpressing LAMP2A improved bone healing, as demonstrated by microcomputed tomography and histological analyses. In summary, LAMP2A positively regulates mMSC osteogenesis and suppresses adipo-osteogenesis, probably via Wnt/beta-catenin/GSK3beta signaling. LAMP2A promoted fracture-healing in the mouse model of tibial fracture. KEY MESSAGES: * LAMP2 positively regulates the mBMSCs osteogenic differentiation. * LAMP2 negatively regulates the mBMSCs adipogenic differentiation. * LAMP2 regulates mBMSCs osteogenesis via Wnt/beta-catenin/GSK3beta signaling pathway. * LAMP2 overexpression mBMSCs promote the fracture healing. CI - (c) 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature. FAU - Wang, Yibo AU - Wang Y AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Hang, Kai AU - Hang K AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Ying, Li AU - Ying L AD - Department of Orthopedic, Taizhou Hospital of Zhejiang Province, Wenzhou Medical University, No. 150, Ximen Street, Linhai, 317000, China. FAU - Wu, Jiaqi AU - Wu J AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Wu, Xiaoyong AU - Wu X AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Zhang, Weijun AU - Zhang W AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Li, Lijun AU - Li L AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Wang, Zhongxiang AU - Wang Z AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Bai, Jinwu AU - Bai J AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. FAU - Gao, Xiang AU - Gao X AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. gaoxiang162@zju.edu.cn. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. gaoxiang162@zju.edu.cn. FAU - Xue, Deting AU - Xue D AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. blueskine@zju.edu.cn. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. blueskine@zju.edu.cn. FAU - Pan, Zhijun AU - Pan Z AUID- ORCID: 0000-0002-4618-5990 AD - Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. zrpzj@zju.edu.cn. AD - Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. zrpzj@zju.edu.cn. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20230510 PL - Germany TA - J Mol Med (Berl) JT - Journal of molecular medicine (Berlin, Germany) JID - 9504370 RN - 3493-12-7 (methionylmethylsulfonium chloride) RN - 0 (Core Binding Factor Alpha 1 Subunit) RN - 0 (Lysosomal Membrane Proteins) RN - 0 (beta Catenin) RN - EC 2.7.11.1 (Glycogen Synthase Kinase 3 beta) SB - IM MH - Mice MH - Animals MH - *Osteogenesis/genetics MH - Core Binding Factor Alpha 1 Subunit/metabolism MH - Lysosomal Membrane Proteins/metabolism MH - beta Catenin/genetics/metabolism MH - Glycogen Synthase Kinase 3 beta/metabolism MH - X-Ray Microtomography MH - Osteoblasts/metabolism MH - *Mesenchymal Stem Cells/metabolism MH - Cell Differentiation/physiology MH - Signal Transduction MH - Wnt Signaling Pathway MH - Cells, Cultured OTO - NOTNLM OT - Adipogenesis OT - CMA OT - LAMP2A OT - Osteogenesis OT - Wnt/beta-catenin/GSK3beta OT - mMSCs EDAT- 2023/05/10 12:42 MHDA- 2023/06/29 06:43 CRDT- 2023/05/10 11:06 PHST- 2022/10/19 00:00 [received] PHST- 2023/04/25 00:00 [accepted] PHST- 2023/04/24 00:00 [revised] PHST- 2023/06/29 06:43 [medline] PHST- 2023/05/10 12:42 [pubmed] PHST- 2023/05/10 11:06 [entrez] AID - 10.1007/s00109-023-02328-1 [pii] AID - 10.1007/s00109-023-02328-1 [doi] PST - ppublish SO - J Mol Med (Berl). 2023 Jul;101(7):783-799. doi: 10.1007/s00109-023-02328-1. Epub 2023 May 10.