The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation

Abstract Background Cytokine licensing with pro-inflammatory molecules, such as tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), has emerged as a promising strategy to enhance the therapeutic potential of multipotent mesenchymal stromal cells (MSCs). While licensing has demonstrate...

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Main Authors: Olena Rogulska, Eliska Vavrinova, Irena Vackova, Jarmila Havelkova, Klara Gotvaldova, Pavel Abaffy, Sarka Kubinova, Michal Sima, Pavel Rossner, Lucie Bacakova, Pavla Jendelova, Katarina Smolkova, Yuriy Petrenko
Format: Article
Language:English
Published: BMC 2025-04-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-025-04309-2
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author Olena Rogulska
Eliska Vavrinova
Irena Vackova
Jarmila Havelkova
Klara Gotvaldova
Pavel Abaffy
Sarka Kubinova
Michal Sima
Pavel Rossner
Lucie Bacakova
Pavla Jendelova
Katarina Smolkova
Yuriy Petrenko
author_facet Olena Rogulska
Eliska Vavrinova
Irena Vackova
Jarmila Havelkova
Klara Gotvaldova
Pavel Abaffy
Sarka Kubinova
Michal Sima
Pavel Rossner
Lucie Bacakova
Pavla Jendelova
Katarina Smolkova
Yuriy Petrenko
author_sort Olena Rogulska
collection DOAJ
description Abstract Background Cytokine licensing with pro-inflammatory molecules, such as tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), has emerged as a promising strategy to enhance the therapeutic potential of multipotent mesenchymal stromal cells (MSCs). While licensing has demonstrated benefits for immunomodulation, its effects on other key MSC functions, including differentiation and paracrine activity, remain incompletely explored. In this study, we evaluated the transcriptomic, metabolomic, and functional changes induced by short-term TNF-α/IFN-γ priming of Wharton’s jelly-derived MSCs (WJ-MSCs). Methods WJ-MSCs were expanded and exposed to TNF-α and IFN-γ (10 ng/ml each) for 24 h. Transcriptomic analysis was performed using RNA sequencing to identify differentially expressed genes related to immune modulation and lineage commitment. Metabolomic profiling was conducted using high-resolution mass spectrometry to assess changes in metabolic pathways. Functional assays evaluated the effects of cytokine priming on induced differentiation and growth factor secretion. Results Cytokine licensing induced notable alterations in gene expression, upregulating pathways linked to immune response, inflammation, and cytokine signalling. However, short-term cytokine treatment significantly attenuated the osteogenic and adipogenic differentiation of MSCs, as evidenced by the reduced expression of RUNX2, ALP, CEBPA, and PPARG. The priming had a negligible effect on EGF, FGF-2, HGF, LIF, and SCF secretion. The production of VEGF-A and VEGF-C was elevated, although the levels remained low. Metabolomic analysis revealed enhanced kynurenine pathway activity, indicative of increased tryptophan catabolism, accompanied by elevated levels of fatty acids and polyamines. Conclusions Our findings demonstrate that TNF-α/IFN-γ priming reprograms WJ-MSCs by enhancing their immunomodulatory capacity at the expense of differentiation potential. These results highlight the need for tailored strategies to optimize MSC functionality for specific clinical applications.
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spelling doaj-art-2297c7eb9d09438e8475ea51b90a240f2025-08-20T02:32:05ZengBMCStem Cell Research & Therapy1757-65122025-04-0116111710.1186/s13287-025-04309-2The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiationOlena Rogulska0Eliska Vavrinova1Irena Vackova2Jarmila Havelkova3Klara Gotvaldova4Pavel Abaffy5Sarka Kubinova6Michal Sima7Pavel Rossner8Lucie Bacakova9Pavla Jendelova10Katarina Smolkova11Yuriy Petrenko12Department of Neuroregeneration, Institute of Experimental Medicine of the Czech Academy of SciencesDepartment of Neuroregeneration, Institute of Experimental Medicine of the Czech Academy of SciencesLaboratory of Biomaterials and Tissue Engineering, Institute of Physiology of the Czech Academy of SciencesDepartment of Neuroregeneration, Institute of Experimental Medicine of the Czech Academy of SciencesLaboratory of Mitochondrial Physiology, Institute of Physiology of the Czech Academy of SciencesLaboratory of Glial Biology and Omics Technologies, Institute of Biotechnology, Czech Academy of SciencesDepartment of Optical and Biophysical Systems, Institute of Physics of the Czech Academy of SciencesDepartment of Toxicology and Molecular Epidemiology, Institute of Experimental Medicine of the Czech Academy of SciencesDepartment of Toxicology and Molecular Epidemiology, Institute of Experimental Medicine of the Czech Academy of SciencesLaboratory of Biomaterials and Tissue Engineering, Institute of Physiology of the Czech Academy of SciencesDepartment of Neuroregeneration, Institute of Experimental Medicine of the Czech Academy of SciencesLaboratory of Mitochondrial Physiology, Institute of Physiology of the Czech Academy of SciencesDepartment of Neuroregeneration, Institute of Experimental Medicine of the Czech Academy of SciencesAbstract Background Cytokine licensing with pro-inflammatory molecules, such as tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), has emerged as a promising strategy to enhance the therapeutic potential of multipotent mesenchymal stromal cells (MSCs). While licensing has demonstrated benefits for immunomodulation, its effects on other key MSC functions, including differentiation and paracrine activity, remain incompletely explored. In this study, we evaluated the transcriptomic, metabolomic, and functional changes induced by short-term TNF-α/IFN-γ priming of Wharton’s jelly-derived MSCs (WJ-MSCs). Methods WJ-MSCs were expanded and exposed to TNF-α and IFN-γ (10 ng/ml each) for 24 h. Transcriptomic analysis was performed using RNA sequencing to identify differentially expressed genes related to immune modulation and lineage commitment. Metabolomic profiling was conducted using high-resolution mass spectrometry to assess changes in metabolic pathways. Functional assays evaluated the effects of cytokine priming on induced differentiation and growth factor secretion. Results Cytokine licensing induced notable alterations in gene expression, upregulating pathways linked to immune response, inflammation, and cytokine signalling. However, short-term cytokine treatment significantly attenuated the osteogenic and adipogenic differentiation of MSCs, as evidenced by the reduced expression of RUNX2, ALP, CEBPA, and PPARG. The priming had a negligible effect on EGF, FGF-2, HGF, LIF, and SCF secretion. The production of VEGF-A and VEGF-C was elevated, although the levels remained low. Metabolomic analysis revealed enhanced kynurenine pathway activity, indicative of increased tryptophan catabolism, accompanied by elevated levels of fatty acids and polyamines. Conclusions Our findings demonstrate that TNF-α/IFN-γ priming reprograms WJ-MSCs by enhancing their immunomodulatory capacity at the expense of differentiation potential. These results highlight the need for tailored strategies to optimize MSC functionality for specific clinical applications.https://doi.org/10.1186/s13287-025-04309-2Multipotent mesenchymal stromal cellsWharton’s jellyCytokine primingTranscriptomicsMetabolomicsAdipogenic and osteogenic differentiation
spellingShingle Olena Rogulska
Eliska Vavrinova
Irena Vackova
Jarmila Havelkova
Klara Gotvaldova
Pavel Abaffy
Sarka Kubinova
Michal Sima
Pavel Rossner
Lucie Bacakova
Pavla Jendelova
Katarina Smolkova
Yuriy Petrenko
The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
Stem Cell Research & Therapy
Multipotent mesenchymal stromal cells
Wharton’s jelly
Cytokine priming
Transcriptomics
Metabolomics
Adipogenic and osteogenic differentiation
title The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
title_full The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
title_fullStr The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
title_full_unstemmed The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
title_short The role of cytokine licensing in shaping the therapeutic potential of wharton’s jelly MSCs: metabolic shift towards immunomodulation at the expense of differentiation
title_sort role of cytokine licensing in shaping the therapeutic potential of wharton s jelly mscs metabolic shift towards immunomodulation at the expense of differentiation
topic Multipotent mesenchymal stromal cells
Wharton’s jelly
Cytokine priming
Transcriptomics
Metabolomics
Adipogenic and osteogenic differentiation
url https://doi.org/10.1186/s13287-025-04309-2
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