Catholic High-Performance Cell Therapy Center & Department of Medical Life Science, College of Medicine, The Catholic University, Seoul 06591, Republic of Korea.
Chemical & Biological Integrative Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea.
Int J Mol Sci. 2023 May 12;24(10):8639. doi: 10.3390/ijms24108639.
Let-7 miRNAs have pleiotropic cellular functions in cell proliferation, migration, and regenerative processes. Here, we investigate whether the inhibition of let-7 miRNAs with antisense oligonucleotides (ASOs) can be a transient and safe strategy enhancing the therapeutic potential of mesenchymal stromal cells (MSCs) to overcome their limitations in cell therapeutic trials. We first identified major subfamilies of let-7 miRNAs preferentially expressed in MSCs, and efficient ASO combinations against these selected subfamilies that mimic the effects of LIN28 activation. When let-7 miRNAs were inhibited with an ASO combination (anti-let7-ASOs), MSCs exhibited higher proliferation with delayed senescence during the passaging into a culture. They also exhibited increased migration and enhanced osteogenic differentiation potential. However, these changes in MSCs were not accompanied by cell-fate changes into pericytes or the additional acquisition of stemness, but instead occurred as functional changes accompanied by changes in proteomics. Interestingly, MSCs with let-7 inhibition exhibited metabolic reprogramming characterized by an enhanced glycolytic pathway, decreased reactive oxygen species, and lower transmembrane potential in mitochondria. Moreover, let-7-inhibited MSCs promoted the self-renewal of neighboring hematopoietic progenitor cells, and enhanced capillary formation in endothelial cells. These findings together show that our optimized ASO combination efficiently reprograms the MSC functional state, allowing for more efficient MSC cell therapy.
Let-7 miRNAs 在细胞增殖、迁移和再生过程中具有多种细胞功能。在这里，我们研究了用反义寡核苷酸 (ASO) 抑制 let-7 miRNAs 是否可以作为一种短暂且安全的策略，来增强间充质基质细胞 (MSCs) 的治疗潜力，以克服其在细胞治疗试验中的局限性。我们首先鉴定了在 MSCs 中优先表达的 let-7 miRNAs 的主要亚家族，并针对这些选定的亚家族设计了有效的 ASO 组合，模拟 LIN28 激活的效果。当用 ASO 组合（抗 let-7-ASOs）抑制 let-7 miRNAs 时，MSCs 在传代培养过程中表现出更高的增殖能力和延迟衰老。它们还表现出更高的迁移能力和增强的成骨分化潜能。然而，这些 MSCs 的变化并没有伴随着向周细胞的细胞命运变化或额外获得干性，而是作为伴随蛋白质组学变化的功能性变化发生的。有趣的是，具有 let-7 抑制的 MSCs 表现出代谢重编程，特征是增强的糖酵解途径、减少的活性氧和线粒体跨膜电位降低。此外，let-7 抑制的 MSCs 促进了邻近造血祖细胞的自我更新，并增强了内皮细胞中的毛细血管形成。这些发现共同表明，我们优化的 ASO 组合可有效地重编程 MSC 的功能状态，从而实现更有效的 MSC 细胞治疗。
