Exosomes from hypoxic ADSCs ameliorate neuronal damage post spinal cord injury through circ-Wdfy3 delivery and inhibition of ferroptosis

Minghao Shao; Sen Ye; Yanzhen Chen; Changzhang Yu; Wei Zhu

doi:10.1016/j.neuint.2024.105759

Exosomes from hypoxic ADSCs ameliorate neuronal damage post spinal cord injury through circ-Wdfy3 delivery and inhibition of ferroptosis

Neurochem Int. 2024 May 10:177:105759. doi: 10.1016/j.neuint.2024.105759. Online ahead of print.

Authors

Minghao Shao¹, Sen Ye², Yanzhen Chen², Changzhang Yu³, Wei Zhu⁴

Affiliations

¹ Department of Orthopedics, Huashan Hospital Affiliated to Fudan University, Shanghai, China.
² Department of Spine Surgery, Xingguo Hospital Affiliated to Gannan Medical University, No. 699 Wenming Avenue, Xingguo County, Ganzhou, 342400, Jiangxi Province, China.
³ Department of Spine Surgery, Xingguo Hospital Affiliated to Gannan Medical University, No. 699 Wenming Avenue, Xingguo County, Ganzhou, 342400, Jiangxi Province, China; Department of Orthopedics, Huashan Hospital Affiliated to Fudan University, Shanghai, China. Electronic address: 13766371331@139.com.
⁴ Department of Spine Surgery, Xingguo Hospital Affiliated to Gannan Medical University, No. 699 Wenming Avenue, Xingguo County, Ganzhou, 342400, Jiangxi Province, China; Department of Orthopedics, Huashan Hospital Affiliated to Fudan University, Shanghai, China. Electronic address: zhuwei2@huashan.org.cn.

PMID: 38735393
DOI: 10.1016/j.neuint.2024.105759

Abstract

Background: Exosomes generated from adipose-derived mesenchymal stem cells (Exos), and in particular hypoxia-pretreated ADSCs (HExos), possess therapeutic properties that promote spinal cord repair following spinal cord injury (SCI). Nevertheless, the regulatory mechanisms through which HExos exert their effects remain unclear.

Methods: Here, next-generation sequencing (NGS) was utilized to examine abnormal circRNA expression comparing HExos to Exos. Bioinformatics analysis and RNA pulldown assays together with luciferase reporter assays were applied to determine interactions among miRNAs, mRNAs and circRNAs. ELISA and immunofluorescence staining were used to examine inflammatory cytokine levels, apoptosis and ROS deposition in LPS-treated HT-22 cells, respectively. The therapeutic effects of Exos and HExos on a mouse model of SCI were analyzed by immunohistochemistry and immunofluorescence staining.

Results: Our findings confirmed that HExos have more significant therapeutic influences on decreasing ROS and inflammatory cytokine levels post-SCI than Exos. NGS revealed that circ-Wdfy3 expression levels were significantly higher in HExos than Exos. Downregulation of circ-Wdfy3 led to a decrease in HExo-induced therapeutic effects on spinal cord repair post-SCI, indicating that circ-Wdfy3 has a critical role in the regulation of HExo-mediated protection against SCI. Our bioinformatics, RNA pulldown and luciferase reporter data demonstrated that GPX4 and miR-423-3p were downstream targets of circ-Wdfy3. GPX4 downregulation or miR-423-3p overexpression reversed the protective effects of circ-Wdfy3 on LPS-treated HT-22 cells. Furthermore, overexpression of circ-Wdfy3 led to an in increase in the Exo-induced therapeutic effects on spinal cord repair post-SCI through the inhibition of ferroptosis.

Conclusions: circ-WDfy3-overexpressing Exos promote spinal cord repair post-SCI through mediation of ferroptosis via the miR-138-5p/GPX4 pathway.

Keywords: ADSCs; Exosomes; Ferroptosis; SCI; circ-Wdfy3.