miRNA-148a–containing GMSC-derived EVs modulate Treg/Th17 balance via IKKB/NF-κB pathway and treat a rheumatoid arthritis model
Jingrong Chen, Xiaoyi Shi, Yanan Deng, Junlong Dang, Yan Liu, Jun Zhao, Rongzhen Liang, Donglan Zeng, Wenbin Wu, Yiding Xiong, Jia Yuan, Ye Chen, Julie Wang, Weidong Lin, Xiangfang Chen, Weishan Huang, Nancy Olsen, Yunfeng Pan, Qingling Fu, Song Guo Zheng
Jingrong Chen, Xiaoyi Shi, Yanan Deng, Junlong Dang, Yan Liu, Jun Zhao, Rongzhen Liang, Donglan Zeng, Wenbin Wu, Yiding Xiong, Jia Yuan, Ye Chen, Julie Wang, Weidong Lin, Xiangfang Chen, Weishan Huang, Nancy Olsen, Yunfeng Pan, Qingling Fu, Song Guo Zheng
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Research Article Stem cells

miRNA-148a–containing GMSC-derived EVs modulate Treg/Th17 balance via IKKB/NF-κB pathway and treat a rheumatoid arthritis model

Abstract

Mesenchymal stem cells (MSCs) have demonstrated potent immunomodulatory properties that have shown promise in the treatment of autoimmune diseases, including rheumatoid arthritis (RA). However, the inherent heterogeneity of MSCs triggered conflicting therapeutic outcomes, raising safety concerns and limiting their clinical application. This study aimed to investigate the potential of extracellular vesicles derived from human gingival mesenchymal stem cells (GMSC-EVs) as a therapeutic strategy for RA. Through in vivo experiments using an experimental RA model, our results demonstrate that GMSC-EVs selectively homed to inflamed joints and recovered Treg and Th17 cell balance, resulting in the reduction of arthritis progression. Our investigations also uncovered miR-148a-3p as a critical contributor to the Treg/Th17 balance modulation via IKKB/NF-κB signaling orchestrated by GMSC-EVs, which was subsequently validated in a model of human xenograft versus host disease (xGvHD). Furthermore, we successfully developed a humanized animal model by utilizing synovial fibroblasts obtained from patients with RA (RASFs). We found that GMSC-EVs impeded the invasiveness of RASFs and minimized cartilage destruction, indicating their potential therapeutic efficacy in the context of patients with RA. Overall, the unique characteristics — including reduced immunogenicity, simplified administration, and inherent ability to target inflamed tissues — position GMSC-EVs as a viable alternative for RA and other autoimmune diseases.

Authors

Jingrong Chen, Xiaoyi Shi, Yanan Deng, Junlong Dang, Yan Liu, Jun Zhao, Rongzhen Liang, Donglan Zeng, Wenbin Wu, Yiding Xiong, Jia Yuan, Ye Chen, Julie Wang, Weidong Lin, Xiangfang Chen, Weishan Huang, Nancy Olsen, Yunfeng Pan, Qingling Fu, Song Guo Zheng

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Figure 6

miR-148a-3p–containing human GMSC–derived EVs modulate IKKB/NF-κB signaling pathway.

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miR-148a-3p–containing human GMSC–derived EVs modulate IKKB/NF-κB signal...
(A) Sequence alignment of miR-148a-3p and its putative target sites in the 3′ UTR of IKKB mRNA. Mutation was generated in the complementary sites for the seed region of miR-148a-3p, as indicated. (B) HEK293T cells were transiently cotransfected with IKKB WT or mutant 3′ UTR luciferase reporter plasmid and miR-148a-3p mimic for 48 hours, and luciferase activity was analyzed. (C and D) HEK293T cells were transiently transfected with negative control or miR-148a-3p mimic. Cells were collected at 48 hours, and the expression of IKKB or p-IKKB was detected by qPCR or Western blot respectively. (E and F) CD3+ T cells isolated from C57BL/6 mice were cocultured with NC-GMSC-EVs or si-GMSC-EVs under the activated condition. Cells were collected at 72 hours, and the expression of IKKB, p-IKKB, and p-NF-κB was detected by qPCR and Western blot. Statistical significance was assessed by 1-way ANOVA with Dunnett multiple-comparison test in C–F and by 2-tailed Student’s t test in B. Data are shown as the means ± SD from 1 of 3 independent experiments. *P < 0.05; **P < 0.01.