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Updated: Jun 9, 2025
In Vivo Imaging of Reactive Oxygen Species in a Murine Wound Model
Published on: November 17, 2018
Non-electrophilic NRF2 activators promote wound healing in human keratinocytes and diabetic mice and demonstrate
May Barakat1,2, Chen Han1,2, Lin Chen1
1Center for Wound Healing and Tissue Regeneration, University of Illinois Chicago College of Dentistry, Chicago, IL, USA.
Abstract:
The transcription factor NRF2 plays an important role in many biological processes and is a promising therapeutic target for many disease states. NRF2 is highly expressed in the skin and is known to play a critical role in diabetic wound healing, a serious disease process for which treatment options are limited. However, many existing NRF2 activators display off-target effects due to their electrophilic mechanism, underscoring the need for alternative approaches. In this work, we investigated two recently described non-electrophilic NRF2 activators, ADJ-310 and PRL-295, and demonstrated their efficacy in vitro and in vivo in human keratinocytes and Leprdb/db diabetic mice. We also compared the downstream targets of PRL-295 to those of the widely used electrophilic NRF2 activator CDDO-Me by RNA sequencing. Both ADJ-310 and PRL-295 maintained human keratinocyte cell viability at increasing concentrations and maintained or improved cell proliferation over time. Both compounds also increased cell migration, improving in vitro wound closure. ADJ-310 and PRL-295 enhanced the oxidative stress response in vitro, and RNA-sequencing data showed that PRL-295 activated NRF2 with a narrower transcriptomic effect than CDDO-Me. In vivo, both ADJ-310 and PRL-295 improved wound healing in Leprdb/db diabetic mice and upregulated known downstream NRF2 target genes in treated tissue. These results highlight the non-electrophilic compounds ADJ-310 and PRL-295 as effective, innovative tools for investigating the function of NRF2. These compounds directly address the need for alternative NRF2 activators and offer a new approach to studying the role of NRF2 in human disease and its potential as a therapeutic across multiple disease states.
Insights
New non-electrophilic activators, ADJ-310 and PRL-295, effectively promote diabetic wound healing by enhancing the NRF2 pathway without off-target effects, offering a promising therapeutic avenue.
Area of Science:
- Biochemistry
- Dermatology
- Pharmacology
Background:
- Nuclear factor erythroid 2-related factor 2 (NRF2) is crucial for biological processes and a therapeutic target for diseases.
- NRF2 is vital in skin biology and diabetic wound healing, but existing activators have off-target effects.
- Diabetic wound healing requires novel therapeutic strategies due to limited treatment options.
Purpose of the Study:
- To investigate the efficacy of novel non-electrophilic NRF2 activators, ADJ-310 and PRL-295.
- To compare the transcriptomic effects of a non-electrophilic activator (PRL-295) with an electrophilic one (CDDO-Me).
- To evaluate the potential of ADJ-310 and PRL-295 in promoting diabetic wound healing.
Main Methods:
- In vitro studies using human keratinocytes to assess cell viability, proliferation, and migration.
- In vivo studies using Leprdb/db diabetic mice to evaluate wound healing.
- RNA sequencing to compare downstream NRF2 targets of PRL-295 and CDDO-Me.
Main Results:
- ADJ-310 and PRL-295 maintained keratinocyte viability and proliferation, and improved in vitro wound closure.
- Both compounds enhanced oxidative stress response in vitro.
- PRL-295 demonstrated a narrower transcriptomic effect compared to CDDO-Me, and both compounds improved wound healing in diabetic mice.
Conclusions:
- Non-electrophilic NRF2 activators ADJ-310 and PRL-295 are effective in promoting diabetic wound healing.
- These compounds offer a safer alternative to electrophilic NRF2 activators, reducing off-target effects.
- ADJ-310 and PRL-295 represent innovative tools for NRF2 research and potential therapeutics for various diseases.
