TET-Mediated DNA Demethylation is Required for Inducible Vasculogenic Reprogramming of the Skin

Tissue nanotransfection (TNT) based topical electrophoretic delivery of Etv2, Foxc2, and Fli1 (EFF) plasmids achieve induced vasculogenesis (iV) in the skin by inducing vasculogenic fibroblasts (VF). scRNA-seq analysis revealed TET-dependent gain of VF in dermal fibroblasts in response to EFF. Emergence of VF required TET-dependent demethylation of fibroblast-borne endothelial genes in vivo. Fibroblast-specific inducible overexpression of EFF in TARGATT mice caused induction of TET1/2/3, followed by transition of fibroblast to VF in vivo. TET induction was associated with elevated 5-hmC abundance in VF. TET-dependent demethylation of fibroblast-borne endothelial genes caused the formation of VF and rescued perfusion in diabetic ischemic limbs. TNTEFF rescued perfusion and closure of diabetic wounds. TET enzymes, suppressed in diabetics, play a pivotal role in causing endogenous EFF-induced VF state change of skin fibroblasts. TNT delivery of EFF transcription factors upregulate TET expression. This work is the first to recognize the significance of TET in the inducible formation of blood vessels to rescue diabetic ischemic tissue. Primary Dermal Fibroblast; Normal, Human, Adult (ATCC ® PCS-201-012) were used.