Journal: Regenerative Therapy
Article Title: IFNγ and TNFα optimize salivary gland mesenchymal stromal cells: an alternative to marrow- and adipose-MSCs for radiation xerostomia
doi: 10.1016/j.reth.2025.11.004
Figure Lengend Snippet: MSC(SG), particularly after cytokine pre-licensing with IFNγ and TNFα, are a feasible and enhanced MSC source for treatment of radiation-induced xerostomia. Bulk RNA sequencing of MSC(BM), MSC(AD), and MSC(SG) revealed that they shared 85 % of transcripts. Key differences included extracellular matrix production and response to cytokines in MSC(SG). Regardless of MSC source, dual stimulation of MSCs with IFNγ and TNFα produced an average of more than a 20-fold increase in R-Spondin 3 compared to vehicle conditions. Additionally, IFNγ and TNFα pre-licensing optimized immunomodulatory marker expression more than IFNγ alone. Intercellular adhesion molecule 1 (ICAM-1) increased 12-fold more, programmed death ligand 1 (PD-L1) increased 1.4-fold more, and indoleamine 2,3 dioxygenase (IDO) increased 2-fold more with IFNγ/TNFα pre-licensing than IFNγ alone. Both cytokine stimulation conditions resulted in a 1.2-fold decrease in T-cell proliferation. Gland structure and salivary flow are preserved in irradiated mice treated with MSC(SG) pre-licensed with IFNγ/TNFα. B) proposed mechanism of MSC action. R-Spondin 3 and Wnt2b generated by MSC(SG) promote epithelial regeneration after damage from radiation. Radiation also creates a pro-inflammatory local environment with increased dual positive T cells. MSC(SG) might protect local epithelial through optimizing the local inflammatory milieu like protecting macrophages that are important for tissue repair. McCoy, S. (2025) https://BioRender.com/o01d792 . Created in https://BioRender.com .
Article Snippet: After 48 h of culture, the conditioned media was used for ELISA per the manufacturer's recommendations (GDNF [Innovative Research Novi MI], RSPO1 [CUSABIO, Houston, TX], RSPO3 [Innovative Research], Wnt1 [CUSABIO], Wnt2b [CUSABIO], Wnt3a [CUSABIO], Wnt4 [Raybiotech, Norcross, GA], Wnt5a [CUSABIO]) with dilutions described in the Supplemental Methods.
Techniques: RNA Sequencing, Produced, Marker, Expressing, Irradiation, Generated