Abstract
Mesenchymal stromal cell (MSC) therapy is making its way into clinical practice, accompanied by research into strategies improving their therapeutic potential. Preconditioning MSCs with hypoxia-inducible factors-α (HIFα) stabilizers is an alternative to hypoxic priming, but there remains insufficient data evaluating its transcriptomic effect. Herein, we determined the gene expression profile of 6 human bone marrow-derived MSCs preconditioned for 6 h in 2% O2 (hypoxia) or with 40 μM Vadadustat, compared to control cells and each other. RNA-Sequencing was performed using the Illumina platform, quality control with FastQC and adapter-trimming with BBDUK2. Transcripts were mapped to the Homo_sapiens. GRCh37 genome and converted to relative expression using Salmon. Differentially expressed genes (DEGs) were generated using DESeq2 while functional enrichment was performed in GSEA and g:Profiler. Comparison of hypoxia versus control resulted in 250 DEGs, Vadadustat versus control 1071, and Vadadustat versus hypoxia 1770. The terms enriched in both phenotypes referred mainly to metabolism, in Vadadustat additionally to vesicular transport, chromatin modifications and interaction with extracellular matrix. Compared with hypoxia, Vadadustat upregulated autophagic, phospholipid metabolism, and TLR cascade genes, downregulated those of cytoskeleton and GG-NER pathway and regulated 74 secretory factor genes. Our results provide valuable insight into the transcriptomic effects of these two methods of MSCs preconditioning.
Highlights
From the comparative analysis of the results of Vadadustat vs. hypoxia preconditioning by Gene set enrichment analysis (GSEA), and which may affect the therapeutic potential of Mesenchymal stromal cell (MSC), we identified that while Vadadustat induces changes in the expression of genes belonging to different categories of signaling, metabolism, and extracellular regulation, while categories related to immunomodulation were enriched after hypoxia preconditioning
Our study confirmed that exposure of MSCs to hypoxia initiates cellular adaptive processes through regulation of gene expression and that some of the regulated genes fit into common expression patterns among different human cells, but some are largely cell-specific
We have shown that activation of gene expression associated with hypoxia preconditioning can be partially achieved by preconditioning with pharmacological stabilizers of hypoxia-inducible factors-α (HIFα)’s, but the two responses are substantially distinct
Summary
Mesenchymal stromal cells (MSCs) are a promising candidate for widely used cellbased therapy. This therapy utilizes the body’s endogenous regenerative mechanism to repair function and restore physiological homeostasis in a broad spectrum of tissues affected by disease processes. MSCs belong to a population of adult progenitor cells that, in addition to their ability to self-renew and differentiate into cells with several mesenchymal phenotypes, are characterized by their potential to be activated in response to tissue injury to promote regenerative processes, mainly through paracrine activity. MSCs are distinguished by their ability to reduce excessive inflammatory processes and regulate both innate and adaptive immune responses
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