Hirakawa Matthew P, Tjahjono Nikki, Light Yooli K, Celebi Aleyna N, Celebi Nisa N, Chintalapudi Prem, Butler Kimberly S, Branda Steven S, Krishnakumar Raga
Systems Biology Department, Sandia National Laboratories, Livermore, CA 94551, USA.
Molecular and Microbiology Department, Sandia National Laboratories, Albuquerque, NM 87185, USA.
iScience. 2022 Jan 12;25(2):103759. doi: 10.1016/j.isci.2022.103759. eCollection 2022 Feb 18.
Mesenchymal stromal cells (MSCs) have broad-ranging therapeutic properties, including the ability to inhibit bacterial growth and resolve infection. However, the genetic mechanisms regulating these antibacterial properties in MSCs are largely unknown. Here, we utilized a systems-based approach to compare MSCs from different genetic backgrounds that displayed differences in antibacterial activity. Although both MSCs satisfied traditional MSC-defining criteria, comparative transcriptomics and quantitative membrane proteomics revealed two unique molecular profiles. The antibacterial MSCs responded rapidly to bacterial lipopolysaccharide (LPS) and had elevated levels of the LPS co-receptor CD14. CRISPR-mediated overexpression of endogenous CD14 in MSCs resulted in faster LPS response and enhanced antibacterial activity. Single-cell RNA sequencing of CD14-upregulated MSCs revealed a shift in transcriptional ground state and a more uniform LPS-induced response. Our results highlight the impact of genetic background on MSC phenotypic diversity and demonstrate that overexpression of CD14 can prime these cells to be more responsive to bacterial challenge.
间充质基质细胞（MSCs）具有广泛的治疗特性，包括抑制细菌生长和消除感染的能力。然而，调节MSCs中这些抗菌特性的遗传机制在很大程度上尚不清楚。在这里，我们采用了一种基于系统的方法来比较来自不同遗传背景、具有不同抗菌活性的MSCs。尽管两种MSCs都满足传统的MSCs定义标准，但比较转录组学和定量膜蛋白质组学揭示了两种独特的分子特征。具有抗菌能力的MSCs对细菌脂多糖（LPS）反应迅速，且LPS共受体CD14水平升高。CRISPR介导的MSCs内源性CD14过表达导致更快的LPS反应和增强的抗菌活性。对CD14上调的MSCs进行单细胞RNA测序揭示了转录基态的转变和更一致的LPS诱导反应。我们的结果突出了遗传背景对MSCs表型多样性的影响，并证明CD14的过表达可以使这些细胞对细菌挑战更敏感。
