Paracrine factors of stressed peripheral blood mononuclear cells activate pro-angiogenic and anti- proteolytic processes in whole blood cells and protect the endothelial barrier

Abstract

Tissue regenerative properties have been attributed to secreted paracrine factors derived from stem cells and other cell types. Especially, the secretome of {gamma}-irradiated peripheral blood mononuclear cells (PBMCsec) has been shown to possess high tissue-regenerative and pro-angiogenic capacities in a variety of preclinical studies. In the light of future therapeutic intravenous applications of PBMCsec, we investigated possible effects of PBMCsec on circulating white blood cells and endothelial cells lining the vasculature. MethodsTo identify changes in the transcriptional profile of white blood cells treated with PBMCSec, whole blood was drawn from healthy individuals and stimulated with PBMCsec for 8 hours ex vivo before further processing for single cell RNA sequencing (scRNAseq). In addition, we performed in vitro assay to confirm findings arising from the transcriptional profiling. ResultsAddition of PBMCsec to whole blood significantly altered the gene signature of granulocytes (17 genes), T-cells (45 genes), B-cells (72 genes) and most prominently monocytes (322 genes). We detected a strong upregulation of several tissue-regenerative and pro-angiogenic cyto- and chemokines in monocytes, including VEGFA, CXCL1 and CXCL5. Intriguingly, inhibitors of endopeptidase activity, such as SERPINB2, were also strongly induced. Measurement of the trans-endothelial electrical resistance of primary human microvascular endothelial cells revealed a strong barrier-protective effect of PBMCsec after barrier disruption. ConclusionTogether, we show that PBMCsec induces angiogenic and proteolytic processes in the blood and is able to attenuate endothelial barrier damage. These regenerative properties suggest that systemic application of PBMCsec might be a promising novel strategy to restore damaged organs.