Stem Cell Program and Gene Therapy Center, University of California Davis, Sacramento, CA 95817, USA.
Institute of Biotechnology, Federal University of Uberlandia, Uberlandia, MG 38408-100, Brazil.
Int J Mol Sci. 2020 Sep 29;21(19):7194. doi: 10.3390/ijms21197194.
For hundreds of indications, mesenchymal stromal cells (MSCs) have not achieved the expected therapeutic efficacy due to an inability of the cells to reach target tissues. We show that inducing high mannose N-glycans either chemically, using the mannosidase I inhibitor Kifunensine, or genetically, using an shRNA to silence the expression of mannosidase I A1 (MAN1A1), strongly increases the motility of MSCs. We show that treatment of MSCs with Kifunensine increases cell migration toward bone fracture sites after percutaneous injection, and toward lungs after intravenous injection. Mechanistically, high mannose N-glycans reduce the contact area of cells with its substrate. Silencing MAN1A1 also makes cells softer, suggesting that an increase of high mannose N-glycoforms may change the physical properties of the cell membrane. To determine if treatment with Kifunensine is feasible for future clinical studies, we used mass spectrometry to analyze the N-glycan profile of MSCs over time and demonstrate that the effect of Kifunensine is both transitory and at the expense of specific N-glycoforms, including fucosylations. Finally, we also investigated the effect of Kifunensine on cell proliferation, differentiation, and the secretion profile of MSCs. Our results support the notion of inducing high mannose N-glycans in MSCs in order to enhance their migration potential.
对于数百种适应症，间充质基质细胞（MSCs）由于无法到达靶组织而未能达到预期的治疗效果。我们表明，通过化学方法（使用甘露糖酶 I 抑制剂 Kifunensine）或遗传方法（使用沉默甘露糖酶 I A1（MAN1A1）表达的 shRNA）诱导高甘露糖 N-聚糖，可强烈增加 MSCs 的迁移能力。我们表明，用 Kifunensine 处理 MSCs 可增加经皮注射后向骨折部位、静脉注射后向肺部的细胞迁移。从机制上讲，高甘露糖 N-聚糖减少了细胞与其底物的接触面积。沉默 MAN1A1 还使细胞变软，这表明高甘露糖 N-糖型的增加可能会改变细胞膜的物理性质。为了确定 Kifunensine 的治疗是否可行用于未来的临床研究，我们使用质谱法分析了 MSCs 随时间推移的 N-聚糖谱，并证明 Kifunensine 的作用是短暂的，并且以特定的 N-聚糖型为代价，包括岩藻糖基化。最后，我们还研究了 Kifunensine 对 MSCs 增殖、分化和分泌谱的影响。我们的结果支持在 MSCs 中诱导高甘露糖 N-聚糖以增强其迁移能力的观点。
