Mesenchymal stromal cells (MSCs) constitute one of the cell types most frequently used in cell therapy. Although several studies have shown the efficacy of these cells to modulate inflammation in different animal models, the results obtained in human clinical trials have been more modest. Here, we aimed at improving the therapeutic properties of MSCs by inducing a transient expression of two molecules that could enhance two different properties of these cells. With the purpose of improving MSC migration towards inflamed sites, we induced a transient expression of the C-X-C chemokine receptor type 4 (CXCR4). Additionally, to augment the anti-inflammatory properties of MSCs, a transient expression of the anti-inflammatory cytokine, interleukin 10 (IL10), was also induced. Human adipose tissue-derived MSCs were transfected with messenger RNAs carrying the codon-optimized versions of CXCR4 and/or IL10. mRNA-transfected MSCs were then studied, first to evaluate whether the characteristic phenotype of MSCs was modified. Additionally, in vitro and also in vivo studies in an LPS-induced inflamed pad model were conducted to evaluate the impact associated to the transient expression of CXCR4 and/or IL10 in MSCs. Transfection of MSCs with CXCR4 and/or IL10 mRNAs induced a transient expression of these molecules without modifying the characteristic phenotype of MSCs. In vitro studies then revealed that the ectopic expression of CXCR4 significantly enhanced the migration of MSCs towards SDF-1, while an increased immunosuppression was associated with the ectopic expression of IL10. Finally, in vivo experiments showed that the co-expression of CXCR4 and IL10 increased the homing of MSCs into inflamed pads and induced an enhanced anti-inflammatory effect, compared to wild-type MSCs. Our results demonstrate that the transient co-expression of CXCR4 and IL10 enhances the therapeutic potential of MSCs in a local inflammation mouse model, suggesting that these mRNA-modified cells may constitute a new step in the development of more efficient cell therapies for the treatment of inflammatory diseases.

中文翻译：

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CXCR4和IL10瞬时异位表达介导的间充质基质细胞抗炎作用增强

间充质基质细胞（MSCs）是细胞疗法中最常用的细胞类型之一。尽管几项研究表明这些细胞在不同动物模型中调节炎症的功效，但在人类临床试验中获得的结果却较为温和。在这里，我们旨在通过诱导两个分子的瞬时表达来改善MSC的治疗特性，这两个分子可以增强这些细胞的两种不同特性。为了改善MSC向发炎部位的迁移，我们诱导了4型CXC趋化因子受体（CXCR4）的瞬时表达。另外，为了增强MSC的抗炎特性，还诱导了抗炎细胞因子白介素10（IL10）的瞬时表达。用携带密码子优化版本的CXCR4和/或IL10的信使RNA转染人脂肪组织来源的MSC。然后研究了mRNA转染的MSC，首先评估了MSC的特征表型是否被修饰。另外，在LPS诱导的发炎垫模型中进行了体外和体内研究，以评估与MSC中CXCR4和/或IL10的瞬时表达相关的影响。用CXCR4和/或IL10 mRNA转染MSC可以诱导这些分子的瞬时表达，而不会改变MSC的特征表型。然后，体外研究显示CXCR4的异位表达显着增强了MSC向SDF-1的迁移，而免疫抑制的增加与IL10的异位表达有关。最后，体内实验表明，与野生型MSC相比，CXCR4和IL10的共表达增加了MSCs向发炎垫的归巢，并诱导了增强的抗炎作用。我们的结果表明，CXCR4和IL10的瞬时共表达增强了局部炎症小鼠模型中MSC的治疗潜力，表明这些mRNA修饰的细胞可能构成了开发更有效的细胞疗法来治疗MSC的新步骤。炎性疾病。