ALKBH5 demethylates the m <sup>6</sup> A modification of SOCS3 in microglia/macrophages and alleviates neuroinflammation after brain injury

L Lin Cai (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) Y Yuqing Liang (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) X Xiaoyu Li R Runxi Fu (Department of Pediatric Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Institute for Pediatric Research) X Xingyu Niu (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) Y Yuxiao Jin (Department of Neurology, Zhongshan Hospital, Laboratory Animal Center, Ministry of Education Frontiers Center for Brain Science, Fudan University) Y Yuxin Li Y Yuheng Zhang (National Children’s Medical Center, Children’s Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, Ministry of Education Frontiers Center for Brain Science, Ministry of Education Innovative Center for New Drug Development of Immune Inflammatory Diseases, Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University) P Pei Ouyang (National Children’s Medical Center, Children’s Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, Ministry of Education Frontiers Center for Brain Science, Ministry of Education Innovative Center for New Drug Development of Immune Inflammatory Diseases, Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University) C Chen Wang Q Qiuyuan Gong (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) Y Yang Yang L Lai Wei Y Yao Jing (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) D Dianxu Yang (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) Z Zhiming Xu (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine) F Fang Yuan J Jun Ding H Hao Chen B Bo Peng Y Yanxia Rao (Department of Neurology, Zhongshan Hospital, Laboratory Animal Center, Ministry of Education Frontiers Center for Brain Science, Fudan University) H Hengli Tian (Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine)

Abstract

Microglia/macrophage-induced neuroinflammation plays a crucial role in the progression of traumatic brain injury (TBI). However, the involvement of N6-methyladenosine (m 6 A) RNA modifications in this process remains elusive. Single-cell RNA sequencing (scRNA-seq) and m 6 A RNA immunoprecipitation sequencing (MeRIP-seq) across multiple time points postinjury revealed a strong correlation between m 6 A modifications and genes enriched in microglia/macrophages. Furthermore, the m 6 A demethylase ALKBH5 was identified as a key regulator of dynamic m 6 A patterns at the injury site. ALKBH5 suppression in microglia/macrophages exacerbated neuroinflammation in vitro and worsened neurological deficits in controlled cortical impact (CCI) models. MeRIP-qPCR and RNA pull-down assays revealed SOCS3 was a downstream target of ALKBH5-mediated m 6 A demethylation. This demethylation stabilized Socs3 mRNA and enhanced its protein expression, which in turn suppressed neuroinflammation via inhibiting the JAK2-STAT3 pathway. Conversely, SOCS3 depletion impaired functional recovery after injury. These findings unveiled a critical ALKBH5–m 6 A–SOCS3 regulatory axis that mitigated microglia/macrophage-driven neuroinflammation after TBI, underscoring its potential as a therapeutic intervention target for TBI progression.

Article Details

Volume / Issue Vol. 122, Issue 39
Published September 30, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (22)

L

Lin Cai

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

Y

Yuqing Liang

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

X

Xiaoyu Li

R

Runxi Fu

Department of Pediatric Surgery, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Institute for Pediatric Research

X

Xingyu Niu

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

Y

Yuxiao Jin

Department of Neurology, Zhongshan Hospital, Laboratory Animal Center, Ministry of Education Frontiers Center for Brain Science, Fudan University

Y

Yuxin Li

Y

Yuheng Zhang

National Children’s Medical Center, Children’s Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, Ministry of Education Frontiers Center for Brain Science, Ministry of Education Innovative Center for New Drug Development of Immune Inflammatory Diseases, Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University

P

Pei Ouyang

National Children’s Medical Center, Children’s Hospital, Institute for Translational Brain Research, State Key Laboratory of Medical Neurobiology, Ministry of Education Frontiers Center for Brain Science, Ministry of Education Innovative Center for New Drug Development of Immune Inflammatory Diseases, Shanghai Key Laboratory of Gene Editing and Cell Therapy for Rare Diseases, Fudan University

C

Chen Wang

Q

Qiuyuan Gong

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

Y

Yang Yang

L

Lai Wei

Y

Yao Jing

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

D

Dianxu Yang

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

Z

Zhiming Xu

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine

F

Fang Yuan

J

Jun Ding

H

Hao Chen

B

Bo Peng

Y

Yanxia Rao

Department of Neurology, Zhongshan Hospital, Laboratory Animal Center, Ministry of Education Frontiers Center for Brain Science, Fudan University

H

Hengli Tian

Department of Neurosurgery, Shanghai Sixth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine