PT - JOURNAL ARTICLE AU - Swathi Balaji AU - Emily Steen AU - Xinyi Wang AU - Hima V. Vangapandu AU - Natalie Templeman AU - Alexander J. Blum AU - Chad M. Moles AU - Hui Li AU - Daria A. Narmoneva AU - Timothy M. Crombleholme AU - Manish J. Butte AU - Paul L. Bollyky AU - Sundeep G. Keswani TI - IL-10 Promotes Endothelial Progenitor Cell Driven Wound Neovascularization and Enhances Healing via STAT3 AID - 10.1101/760165 DP - 2019 Jan 01 TA - bioRxiv PG - 760165 4099 - http://biorxiv.org/content/early/2019/09/08/760165.short 4100 - http://biorxiv.org/content/early/2019/09/08/760165.full AB - Evidence from prior studies of cutaneous trauma, burns, and chronic diabetic wound repair demonstrates that endothelial progenitor cells (EPCs) contribute to de novo angiogenesis, anti-inflammatory reactions, tissue regeneration, and remodeling. We have shown that IL-10, a potent anti-inflammatory cytokine, promotes regenerative tissue repair in an adult model of dermal scar formation via the regulation of fibroblast-specific hyaluronan synthesis in a STAT3 dependent manner. While it is known that IL-10 drives EPC recruitment and neovascularization after myocardial infarction, its specific mode of action, particularly in dermal wound healing and neovascularization in both control and diabetic wounds remains to be defined. Here we show that IL-10 promotes EPC recruitment into the dermal wound microenvironment to facilitate neovascularization and wound healing of control and diabetic (db/db) wounds via vascular endothelial growth factor (VEGF) and stromal-cell derived factor 1 (SDF-1α) signaling. Inducible skin-specific STAT3 knockout (KO) mice were studied to determine whether the impact of IL-10 on the neovascularization and wound healing is STAT3 dependent. We found that IL-10 treatment significantly promotes dermal wound healing with enhanced wound closure, robust granulation tissue formation and neovascularization. This was associated with elevated wound EPC counts as well as increased VEGF and high SDF-1α levels in control mice, an effect that was abrogated in STAT3 KO transgenic mice. These findings were supported in vitro, wherein IL-10-enhanced VEGF and SDF-1α synthesis in primary murine dermal fibroblasts. IL-10-conditioned fibroblast media was shown to promote sprouting and network formation in aortic ring assays. We conclude that overexpression of IL-10 in the wound-specific milieu recruits EPCs and promote neovascularization, which occurs in a STAT3-dependent manner via regulation of VEGF and SDF-1α levels. Collectively, our studies demonstrate that IL-10 increases EPC recruitment leading to enhanced neovascularization and healing of dermal wounds.ΔΔCtComparative Ct4-OHT4-hydroxy tamoxifenAFBAdult FibroblastsANOVAAnalysis of VarianceBGSBovine Growth SerumBMBone marrowCD34cluster of differentiation 34 moleculeCD133cluster of differentiation 133 molecule (also known as Prominin1)CXCR4CXC chemokine receptor type 4DMEMDulbecco’s Modified Eagle’s MediaECLEnhanced ChemoluminescenceECsEndothelial cellsECMExtracellular MatrixELISAEnzyme-Linked Immunosorbent AssayEPCsEndothelial progenitor cellsFLK-1Fetal Liver Kinase 1 (also known as KDR, is a receptor for VEGF)GFPGreen Fluorescent ProteinH&EHematoxylin and EosinHIF-1Hypoxia inducible factor 1HSCsHematopoietic stem cellsIgGImmunoglobulin GIHCImmunohistochemistryIL-10Interleukin 10IL-10RInterleukin 10 ReceptorLV-GFPLenti Virus expressing GFPLV-IL-10Lenti Virus expressing IL-10MIMyocardial InfarctionPBSPhosphate Buffered SalinePSFPenicillin, Streptomycin, Amphotericinp-STAT3Phosphorylated Signal Transducer and Activator of Transcription 3qPCRReal Time-Polymerase Chain ReactionSDF-1αStromal cell-derived factor 1αshRNAShort Hairpin RNASTAT3Signal Transducer and Activator of Transcription 3TBSTris-Buffered Saline\TGF-β1, β3Transforming Growth Factor-β1, β3Tween-20Polysorbate-20VEGFVascular endothelial growth factor