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Hypoxia- and inflammation-driven preconditioning modulates angiogenic and metabolic pathways in canine
Pablo Ocampo-Ortiz1, Viviana Vallejo-Aristizabal2, Marcos Gomides Carvalho3
1Department of veterinary Medicine, Faculty of Agricultural Sciences, University of Pamplona, Pamplona, Norte de Santander, Colombia.
Abstract:
The immunomodulatory properties of exogenous mesenchymal stem cells (MSCs) have been the target of research in immune-mediated diseases and organ transplants. However, the altered microenvironment decrease MSCs capabilities and survival post-transplantation. This study investigated the viability, proliferation, gene expression and proteomic of canine adipose tissue-derived MSCs (cAT-MSCs) treated with deferroxyamine [DFO] (hypoxia), interferon-γ [IFN-γ] (inflammation) or both for 48h. At 24 hours, all groups exhibited fibroblastoid morphology and adhesion to plastic, with treated groups showing greater cell spacing. After 144h, cell proliferation did not differ significantly between groups, though the treated groups had higher cell concentrations compared to the control. Gene expression analysis revealed increased Casp9 expression in the IFN-γ group, in comparison to the IFN-γ + DFO group; the FGF2 gene was upregulated in the IFN-γ group, while the DKC1 and PT53 genes showed higher expression in IFN-γ than DFO. The VEGFA was more highly expressed in the groups treated with DFO. Proteomics analysis identified 256 proteins, with 70 co-expressed across all groups, and unique proteins in each treatment group: 41 in the control, 44 in DFO, 15 in IFN-γ + DFO group, and 34 for IFN-γ. Notably, 6, 5, and 4 proteins were unique to DFO, IFN-γ + DFO, and IFN-γ treatments, respectively, when compared to the control. Preconditioning modulated angiogenic and metabolic pathways, preserving immunomodulatory function and cellular integrity. Future studies with real hypoxia and multi-omics integration will be crucial for linking molecular signatures to paracrine functions and in vivo efficacy.
Insights
Preconditioning canine adipose-derived mesenchymal stem cells (cAT-MSCs) with deferroxyamine (hypoxia) or interferon-gamma (inflammation) improved cell concentration and modulated angiogenic pathways. These findings suggest potential for enhancing MSC efficacy in regenerative medicine applications.
Area of Science:
- Stem Cell Biology
- Immunomodulation
- Regenerative Medicine
Background:
- Exogenous mesenchymal stem cells (MSCs) show promise for immune-mediated diseases and organ transplants.
- The transplant microenvironment can impair MSC capabilities and survival.
- Investigating preconditioning strategies is crucial to enhance MSC efficacy.
Purpose of the Study:
- To evaluate the impact of deferroxyamine (DFO) and interferon-gamma (IFN-γ) preconditioning on canine adipose tissue-derived MSCs (cAT-MSCs).
- To assess changes in cAT-MSC viability, proliferation, gene expression, and proteomic profiles.
- To understand how DFO and IFN-γ treatments affect MSCs in simulated inflammatory and hypoxic conditions.
Main Methods:
- Canine adipose tissue-derived MSCs (cAT-MSCs) were treated with DFO (hypoxia), IFN-γ (inflammation), or both for 48 hours.
- Assessed cell morphology, adhesion, proliferation, and cell concentration at 24 and 144 hours.
- Performed gene expression analysis (Casp9, FGF2, DKC1, PT53, VEGFA) and proteomics analysis.
Main Results:
- Pretreated groups showed greater cell spacing at 24 hours and higher cell concentrations at 144 hours compared to controls.
- Gene expression varied, with increased Casp9 in IFN-γ group vs. IFN-γ+DFO; VEGFA upregulated with DFO treatment.
- Proteomics identified unique protein profiles for each treatment group, revealing modulation of angiogenic and metabolic pathways.
Conclusions:
- DFO and IFN-γ preconditioning can enhance MSC concentration and modulate key cellular pathways.
- Preconditioning appears to preserve immunomodulatory function and cellular integrity under simulated adverse conditions.
- Further research integrating multi-omics and in vivo studies is needed to confirm paracrine functions and therapeutic efficacy.
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