Zheng Yuanjing, Li Yue, Wei Zhengyang, Wang Yang, Liu Yuanlin, Liu Fengsong, Li Xue, Zhang Yi
Key Laboratory of Zoological Systematics and Application, College of Life Sciences, Hebei University, Baoding, 071002, China.
Beijing Institute of Radiation Medicine, Beijing, 100850, China.
Biochem Biophys Rep. 2025 Jun 9;43:102078. doi: 10.1016/j.bbrep.2025.102078. eCollection 2025 Sep.
Nowadays mesenchymal stem cell-derived exosomes (MSC-Exos) have emerged as a promising cell-free therapeutic alternative to MSC-based therapies, demonstrating efficacy in treating degenerative diseases, inflammatory disorders, and autoimmune diseases. MSC-Exos transport bioactive cargoes such as proteins, lipids, mRNAs, and microRNAs (miRNAs) to the recipient cells, mediating intercellular communication to regulate immunomodulation and tissue repair. However, the exosomal miRNA profile varies dynamically based on the culture conditions and tissue sources. Thus, elucidating the specific exosomal miRNA profile and regulatory targets is critical for the precise clinical applications and development of MSC-Exos-based cell-free therapies. Here we established an optimized serum-free culture system for human umbilical cord-derived MSCs (hUC-MSCs) and determined the critical 48-72-h harvest window for exosome secretion. High-throughput sequencing identified miR-16-5p as the predominant exosomal miRNA, functioning as a core immunosuppressive effector by suppressing LPS/IFN-γ-induced M1 macrophage polarization and Th1 cell differentiation. Mechanistically, miR-16-5p was found to target key nodes in NF-κB and JAK-STAT pathways, validated via dual-luciferase assays. Additionally, miR-125b-5p and miR-34a-5p enhanced this immunosuppressive effect by co-targeting overlapping pathway components in NF-κB and JAK-STAT pathways, suggesting a multilayered regulatory network. Taken together, our findings highlight the potential of miRNA-engineered exosomes as standardized therapies for inflammatory disorders, emphasizing the importance of optimizing culture conditions and profiling miRNA expression over time in advancing clinical translation.
如今，间充质干细胞衍生的外泌体（MSC-Exos）已成为一种有前景的无细胞治疗替代方案，可替代基于MSC的疗法，在治疗退行性疾病、炎症性疾病和自身免疫性疾病方面显示出疗效。MSC-Exos将蛋白质、脂质、mRNA和微小RNA（miRNA）等生物活性物质转运至受体细胞，介导细胞间通讯以调节免疫调节和组织修复。然而，外泌体miRNA谱会根据培养条件和组织来源而动态变化。因此，阐明特定的外泌体miRNA谱及其调控靶点对于基于MSC-Exos的无细胞疗法的精确临床应用和开发至关重要。在此，我们建立了一种优化的无血清培养系统用于人脐带间充质干细胞（hUC-MSCs），并确定了外泌体分泌的关键48 - 72小时收获窗口。高通量测序确定miR-16-5p为主要的外泌体miRNA，通过抑制LPS/IFN-γ诱导的M1巨噬细胞极化和Th1细胞分化发挥核心免疫抑制效应。机制上，发现miR-16-5p靶向NF-κB和JAK-STAT通路中的关键节点，通过双荧光素酶测定法得到验证。此外，miR-125b-5p和miR-34a-5p通过共同靶向NF-κB和JAK-STAT通路中重叠的通路成分增强了这种免疫抑制作用，提示存在一个多层调控网络。综上所述，我们的研究结果突出了miRNA工程化外泌体作为炎症性疾病标准化疗法的潜力，强调了优化培养条件和随时间分析miRNA表达在推进临床转化中的重要性。
