The NSUN5-FTH1/FTL pathway mediates ferroptosis in bone marrow-derived mesenchymal stem cells

Jie Liu,Helu Liu,Weihong Kuang,Yi Li,Lin Yang,Zhenxing Ren,Yichun Ji,Lulu Zhu,Meiling Zhu,Caiqun Bie,Dongfeng Chen

doi:10.1038/s41420-022-00902-z

Abstract

Ferroptosis is a type of cell death induced by the iron-dependent accumulation of lipid hydroperoxides and reactive oxygen species (ROS) in cells. Inhibiting ferroptosis is important for improving the survival of transplanted bone marrow-derived mesenchymal stem cells (BMSCs). Although it is known that NOP2/Sun RNA methyltransferase 5 (NSUN5) post-transcriptionally regulates ferroptosis in BMSCs through RNA methylation, the precise mechanisms underlying these effects have not been reported. In this study, we demonstrate that NSUN5 is downregulated in erastin-induced ferroptosis in BMSCs. Ferroptosis was inhibited by the overexpression of NSUN5 or ferritin heavy chain/light-chain (FTH1/FTL) and was enhanced by NSUN5 knockdown. RNA immunoprecipitation experiments revealed that NSUN5 binds to FTH1/FTL, while NSUN5 depletion reduced the levels of 5-methylcytosine in FTH1/FTL RNA and increased intracellular iron concentrations, resulting in the downregulation of glutathione peroxidase 4 (GPX4) and the accumulation of ROS and lipid peroxidation products. Co-immunoprecipitation experiments demonstrated that the recognition of FTH1 and FTL by NSUN5 is dependent on the recruitment of tumor necrosis factor receptor-associated protein 1 (TRAP1). These results suggested that the NSUN5-FTH1/FTL pathway mediates ferroptosis in BMSCs and that the therapeutic targeting of components of this pathway may promote resistance to ferroptosis and improve the survival of transplanted BMSCs.

Highlights

Mesenchymal stem cell (MSC) transplantation is an important treatment option for acute liver failure as well as for cardiac and neurodegenerative diseases [1–3]
MSC transplantation is used for the treatment of several diseases; the ferroptosis of Bone marrow-derived mesenchymal stem cells (BMSCs) can lead to transplant failure, thereby limiting their use
We used erastin treatment to investigate the roles of NSUN5 in BMSC ferroptosis

Summary

INTRODUCTION

Mesenchymal stem cell (MSC) transplantation is an important treatment option for acute liver failure as well as for cardiac and neurodegenerative diseases [1–3]. Of analysis indicated that NSUN5 depletion induced the C-to-T the seven RNA methyltransferases evaluated, only NSUN5 and conversion at FTH1 and FTL mRNA cytosine methylation sites, NSUN7 mRNA expression levels differed significantly between lead to the reduce of the methylate modification These results control and erastin-treated BMSCs (Fig. 2A–H). Immunofluorescence labeling and confocal microscopy analysis revealed that NSUN5 recruits TRAP1 to modify FTH1 and FTL the NSUN5 fluorescence signal was weaker in erastin-treated We investigated whether other proteins are involved in the BMSCs than in control cells (Fig. 2I, J) These results suggested that NSUN5-mediated modification of FTH1 and FTL. TRAP1 is a mitochondrial paralog of with untreated controls, the expression levels of glutathione heat-shock protein 90 (Hsp90) that has been reported to induce peroxidase 4 (GPX4) were lower and higher with NSUN5 knock- cell death and may regulate ferroptosis in BMSCs. We down and overexpression, respectively, in BMSCs treated with erastin (Fig. 3B). The suggested that, in the absence of NSUN5, theL through the results showed that NSUN5 bound FTH1 and FTL mRNA in BMSCs recruitment ointeraction between TRAP1 and FTH1/FTL was

DISCUSSION

MATERIALS AND METHODS

ETHICS APPROVAL