LAMP2A regulates the balance of mesenchymal stem cell adipo-osteogenesis via the Wnt/β-catenin/GSK3β signaling pathway

Yibo Wang; Kai Hang; Li Ying; Jiaqi Wu; Xiaoyong Wu; Weijun Zhang; Lijun Li; Zhongxiang Wang; Jinwu Bai; Xiang Gao; Deting Xue; Zhijun Pan

doi:10.1007/s00109-023-02328-1

LAMP2A regulates the balance of mesenchymal stem cell adipo-osteogenesis via the Wnt/β-catenin/GSK3β signaling pathway

J Mol Med (Berl). 2023 Jul;101(7):783-799. doi: 10.1007/s00109-023-02328-1. Epub 2023 May 10.

Authors

Yibo Wang^#^{1

2}, Kai Hang^#^{1

2}, Li Ying^#³, Jiaqi Wu^{1

2}, Xiaoyong Wu^{1

2}, Weijun Zhang^{1

2}, Lijun Li^{1

2}, Zhongxiang Wang^{1

2}, Jinwu Bai^{1

2}, Xiang Gao^{4

5}, Deting Xue^{6

7}, Zhijun Pan^{8

9}

Affiliations

¹ Department of Orthopedic Surgery of the Second Affiliated Hospital, School of Medicine, Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China.
² Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China.
³ Department of Orthopedic, Taizhou Hospital of Zhejiang Province, Wenzhou Medical University, No. 150, Ximen Street, Linhai, 317000, China.
⁴ 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.
⁵ Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. gaoxiang162@zju.edu.cn.
⁶ 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.
⁷ Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. blueskine@zju.edu.cn.
⁸ 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.
⁹ Orthopedics Research Institute of Zhejiang University, No. 88, Jiefang Road, Hangzhou, 310009, China. zrpzj@zju.edu.cn.

^# Contributed equally.

PMID: 37162558
DOI: 10.1007/s00109-023-02328-1

Abstract

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 Gsk3β. 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 PPARγ and C/EBPα; LAMP2A overexpression had the opposite effects. The active-β-catenin and phospho-GSK3β (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/β-catenin/GSK3β 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/β-catenin/GSK3β signaling pathway. • LAMP2 overexpression mBMSCs promote the fracture healing.

Keywords: Adipogenesis; CMA; LAMP2A; Osteogenesis; Wnt/β-catenin/GSK3β; mMSCs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation / physiology
Cells, Cultured
Core Binding Factor Alpha 1 Subunit / metabolism
Glycogen Synthase Kinase 3 beta / metabolism
Lysosomal Membrane Proteins / metabolism
Mesenchymal Stem Cells* / metabolism
Mice
Osteoblasts / metabolism
Osteogenesis* / genetics
Signal Transduction
Wnt Signaling Pathway
X-Ray Microtomography
beta Catenin / genetics
beta Catenin / metabolism

Substances

methionylmethylsulfonium chloride
Core Binding Factor Alpha 1 Subunit
Lysosomal Membrane Proteins
beta Catenin
Glycogen Synthase Kinase 3 beta