title: Restoration of Nrf2 Signaling Normalizes the Regenerative Niche.
abstract: Chronic hyperglycemia impairs intracellular redox homeostasis and contributes to impaired diabetic tissue regeneration. The Keap1/Nrf2 pathway is a critical regulator of the endogenous antioxidant response system, and its dysfunction has been implicated in numerous pathologies. Here we characterize the effect of chronic hyperglycemia on Nrf2 signaling within a diabetic cutaneous regeneration model. We characterized the effects of chronic hyperglycemia on the Keap1/Nrf2 pathway within models of diabetic cutaneous wound regeneration. We assessed reactive oxygen species (ROS) production and antioxidant gene expression following alterations in the Nrf2 suppressor Keap1 and the subsequent changes in Nrf2 signaling. We also developed a topical small interfering RNA (siRNA)-based therapy to restore redox homeostasis within diabetic wounds. Western blotting demonstrated that chronic hyperglycemia-associated oxidative stress inhibits nuclear translocation of Nrf2 and impairs activation of antioxidant genes, thus contributing to ROS accumulation. Keap1 inhibition increased Nrf2 nuclear translocation, increased antioxidant gene expression, and reduced ROS production to normoglycemic levels, both in vitro and in vivo. Topical siKeap1 therapy resulted in improved regenerative capacity of diabetic wounds and accelerated closure. We report that chronic hyperglycemia weakens the endogenous antioxidant response, and the consequences of this defect are manifested by intracellular redox dysregulation, which can be restored by Keap1 inhibition. Targeted siRNA-based therapy represents a novel, efficacious strategy to reestablish redox homeostasis and accelerate diabetic cutaneous tissue regeneration.
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Marc A
authors.initials: MA
authors.lastname: Soares
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Oriana D
authors.initials: OD
authors.lastname: Cohen
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Yee Cheng
authors.initials: YC
authors.lastname: Low
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Rita A
authors.initials: RA
authors.lastname: Sartor
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Trevor
authors.initials: T
authors.lastname: Ellison
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Utkarsh
authors.initials: U
authors.lastname: Anil
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Lavinia
authors.initials: L
authors.lastname: Anzai
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Jessica B
authors.initials: JB
authors.lastname: Chang
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Pierre B
authors.initials: PB
authors.lastname: Saadeh
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY.
authors.forename: Piul S
authors.initials: PS
authors.lastname: Rabbani
authors.affiliation: Hansjörg Wyss Department of Plastic Surgery, New York University Langone Medical Center, New York, NY daniel.ceradini@nyumc.org.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Ceradini
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D015153:Blotting, Western; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D003920:Diabetes Mellitus / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005978:Glutathione; D006943:Hyperglycemia / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D000072019:Kelch-Like ECH-Associated Protein 1; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D018384:Oxidative Stress / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012038:Regeneration / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D012867:Skin / Q000293:injuries / Q000378:metabolism*; D014945:Wound Healing / Q000502:physiology*; D014947:Wounds and Injuries / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; C117301:Keap1 protein, mouse; D000072019:Kelch-Like ECH-Associated Protein 1; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D017382:Reactive Oxygen Species; D005978:Glutathione