Mesenchymal stromal cells (MSCs) have broad-ranging therapeutic capabilities, however MSC use is confounded by cell-to-cell heterogeneity, and source-to-source phenotypic inconsistencies. We utilized a systems-based approach to compare MSCs which displayed different capacity for antibacterial activity. Although MSCs from both sources satisfied traditional MSC-defining criteria, comparative transcriptomics and quantitative membrane proteomics demonstrated two unique molecular profiles. The antibacterial MSCs respond rapidly to bacterial lipopolysaccharide (LPS) and have elevated levels of the LPS co-receptor CD14. CRISPR-mediated overexpression of endogenous CD14 in non-antibacterial MSCs resulted in faster LPS response and enhanced antimicrobial activity. Single-cell transcriptome profiling of CD14-activated MSCs revealed uniform enhancement of LPS response kinetics, and a shift in the ground state of these MSCs. Our results demonstrate that systems-level analysis can reveal critical molecular targets to optimize desirable properties in MSCs, and that overexpression of CD14 in these cells can shift their state to be more responsive to future bacterial challenge.

中文翻译：

---

通过系统级分析和CRISPR介导的CD14激活增强间质基质细胞的抗菌活性。

间充质基质细胞（MSCs）具有广泛的治疗能力，但是MSC的使用因细胞间异质性和来源间表型不一致而混淆。我们利用基于系统的方法来比较具有不同抗菌活性的MSC。尽管两种来源的MSC都符合传统的MSC定义标准，但比较转录组学和定量膜蛋白质组学显示了两个独特的分子谱。抗菌MSC对细菌脂多糖（LPS）迅速反应，并具有升高水平的LPS共受体CD14。CRISPR介导的非抗菌MSC中内源性CD14的过表达导致更快的LPS反应和增强的抗菌活性。CD14激活的MSCs的单细胞转录组分析显示，LPS反应动力学均匀增强，并且这些MSC的基态发生了变化。我们的结果表明，系统级分析可以揭示关键的分子靶标，以优化MSC中所需的特性，并且这些细胞中CD14的过表达可以改变其状态，从而对未来的细菌攻击做出更大的响应。