NRF2 Activation in Diabetic Wound Healing: Evaluation of Novel NRF2 Activators ADJ-310 and PRL-295

The transcription factor NRF2 is a prominent research target and is known to play a role in many important cellular and disease processes, including redox homeostasis, stress response, metabolism, and many more. To understand the functions of NRF2 and unlock its potential as a therapeutic, many NRF2-activating compounds have been used to investigate it in both pre-clinical and clinical studies. However, many of these NRF2 activators have disadvantages that can cause toxicity and off-target effects. Here, we employ a newly-synthesized family of small molecule NRF2 activators that are specifically designed to reduce the challenges posed by pre-existing activators. In Chapter 1, we introduce our disease of interest - the chronic diabetic wound – and present a literature review of the many research targets that have improved wound healing in pre-clinical studies. We describe the relative lack of treatment options for diabetic foot ulcers, and we introduce NRF2 as a more powerful research target. In Chapter 2, we investigate the effects of two novel NRF2 activators, ADJ-310 and PRL-295, in vitro and in vivo and conduct an mRNA Sequencing analysis to further understand the downstream targets and mechanisms of the NRF2 pathway. Our results demonstrate that ADJ-310 and PRL-295 are effective, promote wound healing functions in keratinocytes, enhance the oxidative stress response, improve diabetic wound healing in vivo, and have reduced off-target effects compared to the pre-existing NRF2 activator, Bardoxolone-methyl, or CDDO-Me. Finally, in Chapter 3, we present a modified husbandry protocol to reduce attrition and improve the reliability and reproducibility of wound healing studies using the Leprdb/db mouse.