中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2025, Vol. 20 ›› Issue (11): 3273-3286.doi: 10.4103/NRR.NRR-D-24-00068

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

坏死性凋亡神经干细胞分泌的外泌体可参与脊髓损伤后的细胞通讯

  

  • 出版日期:2025-11-15 发布日期:2025-02-25

Exosomes originating from neural stem cells undergoing necroptosis participate in cellular communication by inducing TSC2 upregulation of recipient cells following spinal cord injury

Shiming Li1, #, Jianfeng Li1, #, Guoliang Chen2, #, Tao Lin3 , Penghui Zhang1, *, Kuileung Tong4, *, Ningning Chen1, *, Shaoyu Liu5   

  1. 1 Shenzhen Key Laboratory of Bone Tissue Repair and Translational Research, Department of Orthopedic Surgery, The Seventh Affiliated Hospital, Sun Yat-sen University, Shenzhen, Guangdong Province, China;  2 Department of Orthopedic Surgery, The First Affiliated Hospital of Jinan University, Guangzhou, Guangdong Province, China;  3 Department of Orthopedics and Traumatology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong Province, China;  4 Department of Orthopedic Surgery, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China;  5 Department of Orthopedic Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China
  • Online:2025-11-15 Published:2025-02-25
  • Contact: Ningning Chen, PhD, chennn8@mail.sysu.edu.cn; Kuileung Tong, PhD, tongkleung@mail3.sysu.edu.cn; Penghui Zhang, PhD, zphspine@163.com.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, No. 81801907 (to NC); Shenzhen Key Laboratory of Bone Tissue Repair and Translational Research, No. ZDSYS20230626091402006 (to NC); Sanming Project of Medicine in Shenzhen, No. SZSM201911002 (to SL); Foundation of Shenzhen Committee for Science and Technology Innovation, Nos. JCYJ20230807110310021 (to NC), JCYJ20230807110259002 (to JL); and Science and Technology Program of Guangzhou, No. 2024A04J4716 (to TL).

PDF

11

摘要：

作者既往研究已证实抑制神经干细胞的程序性坏死可促进脊髓损伤后的功能恢复。外泌体被认为可参与干细胞外分泌功能，其在脊髓损伤后的作用尚不明确。为探索神经干细胞发生程序性坏死后产生的外泌体在脊髓损伤后发挥的作用，实验首先通过单细胞测序数据印证了神经干细胞来源于室管膜细胞，并可在脊髓损伤后发生程序性坏死。随后，从E16-17胎鼠中提取神经干细胞，构建了体外坏死模型，并提取外泌体，发现坏死对外泌体的基本特性和产量没有明显影响。对上述外泌体进行全转录测序，鉴定出差异表达的108种mRNA、104种长链非编码RNA、720种环状RNA和14种微小RNA。继而构建竞争性内源RNA网络筛选出Tsc2，Slc16a3和Foxp1三种枢纽基因。在体内实验中证实，损伤脊髓组织中TSC2表达上调，且TSC2阳性细胞存在于损伤区SOX2阳性细胞周围。此外，也在体外实验中发现外泌体受体细胞中TSC2表达的增加。最后，对脊髓损伤后细胞通讯的进一步分析显示，Tsc2在脊髓损伤后1和3d通过表皮生长因子和Midkine信号通路参与室管膜细胞通讯，而Slc16a3在脊髓损伤后7d通过血管内皮生长因子和巨噬细胞迁移抑制因子信号通路参与管膜细胞的细胞通讯。上述结果证实，来源于经历了坏死的神经干细胞的外泌体可参与脊髓损伤后的细胞通讯，并能诱导受体细胞中TSC2表达的上调。

https://orcid.org/0000-0002-4576-2311 (Ningning Chen); https://orcid.org/0000-0001-9846-5655 (Kuileung Tong); 
https://orcid.org/0000-0002-3332-4453 (Penghui Zhang); https://orcid.org/0000-0002-7253-0495 (Shiming Li)

关键词: 脊髓损伤, 神经干细胞, 坏死, 外泌体, Tsc2, Slc16a3, Foxp1, 转录组测序, 竞争性内源RNA, 细胞通讯

Abstract: We previously demonstrated that inhibiting neural stem cells necroptosis enhances functional recovery after spinal cord injury. While exosomes are recognized as playing a pivotal role in neural stem cells exocrine function, their precise function in spinal cord injury remains unclear. To investigate the role of exosomes generated following neural stem cells necroptosis after spinal cord injury, we conducted singlecell RNA sequencing and validated that neural stem cells originate from ependymal cells and undergo necroptosis in response to spinal cord injury. Subsequently, we established an in vitro necroptosis model using neural stem cells isolated from embryonic mice aged 16–17 days and extracted exosomes. The results showed that necroptosis did not significantly impact the fundamental characteristics or number of exosomes. Transcriptome sequencing of exosomes in necroptosis group identified 108 differentially expressed messenger RNAs, 104 long non-coding RNAs, 720 circular RNAs, and 14 microRNAs compared with the control group. Construction of a competing endogenous RNA network identified the following hub genes: tuberous sclerosis 2 (Tsc2), solute carrier family 16 member 3 (Slc16a3), and forkhead box protein P1 (Foxp1). Notably, a significant elevation in TSC2 expression was observed in spinal cord tissues following spinal cord injury. TSC2-positive cells were localized around SRY-box transcription factor 2–positive cells within the injury zone. Furthermore, in vitro analysis revealed increased TSC2 expression in exosomal receptor cells compared with other cells. Further assessment of cellular communication following spinal cord injury showed that Tsc2 was involved in ependymal cellular communication at 1 and 3 days post-injury through the epidermal growth factor and midkine signaling pathways. In addition, Slc16a3 participated in cellular communication in ependymal cells at 7 days post-injury via the vascular endothelial growth factor and macrophage migration inhibitory factor signaling pathways. Collectively, these findings confirm that exosomes derived from neural stem cells undergoing necroptosis play an important role in cellular communication after spinal cord injury and induce TSC2 upregulation in recipient cells.

Key words: cellular communication, competing endogenous RNA, exosomes, Foxp1, necroptosis, neural stem cells, Slc16a3, spinal cord injury, transcriptome sequencing, Tsc2