Pro-inflammatory cytokine primed Wharton's jelly mesenchymal stem cells: A potent immunomodulatory strategy to combat pancreatic cancer

Background & Aim: Mesenchymal stem cells (MSCs) from umbilical cord (UC) have the potential therapeutic applications in context of injured, immune related & inflammatory diseases due to high proliferation rate, multilineage differentiation potential and immune tolerance properties. While clinical trials have clearly demonstrated strong safety records, efficacy of MSCs is still modest and inconsistent. Hypothesizing that priming MSCs prior to administration can improve their therapeutic efficacy, present study investigates the modulation of MSC behaviour through cytokine priming and assesses their effectiveness in ameliorating tumor apoptosis. Proteomic analysis shows compositional changes post cytokine priming. Method: Wharton's jelly derived MSCs (hWJMSCs) were primed with tumor necrosis factor-alpha (TNF-alpha; 25ng/ml) and interferon-gamma (IFN-gamma; 25ng/ml), incubated for 24 hrs at 37°C/5%CO2, their morphological alterations, multi-lineage differentiation potentials (osteocytes/chondrocytes/adipocytes), immunophenotypying (CD90+73+105+44+34-45-19- HLADR-), Adhesion molecules (ICAM, VCAM, NCAM) migration capabilities were examined.Label-free quantitative LCMS (nLC/MS/MS coupled to Orbitrap fusion Tribrid mass-spectrometer) analysis was performed on more than 2400 proteins identified. Further, immunogenic potentials are being tested using PBMNCs from healthy human donors. Result: hWJMSCs primed with TNF-alpha (hWJMSC-T)/IFN-gamma (HWJMSC-I) showed >90% expression of CD90 + 73 + 105 + 44 + . Microscopically morphological distinction stained with phyalloidin/DAPI observed showed hWJMSC-T as elongated and flattened cells and Complement & coagulation cascade and Pro-angiogenic proteome signature in abundance, suggesting their role in innate immunity homeostasis. On the other hand, IFN-gamma induction showed immune-modulating protein signature of infection/inflammation especially, Guanylate-binding proteins. Differentially expressed protein signature points towards modulation of NOD-like receptor signaling and cell adhesion molecule pathways. Conclusion: Data herein provides the evidences that stem cell properties of regenerative capability remains conserved and do not compromise under inflammatory conditions. Additionally, primed hWJMSCs can modulate or prevent infection most likely via cross-talk between cascades of defense systems. Thus, suggesting a potential role as alternate strategy to regulate inflammation and/or reshape inflammatory microenvironment. Keywords: Mesenchymal stem cells, pro-inflammatory cytokines, inflammation, immunomodulation