Objectives/Goals: The primary objective of this study is to test whether NAD+ modulation lowers mitochondrial oxidative stress to restore endothelial nitric-oxide signaling and limit microvascular remodeling in diabetes in murine and 3D tissue-engineered models. The goal is to define NAD-responsive pathways and candidate blood biomarkers of microvascular injury. Methods/Study Population: Use db/db diabetic mice to measure effects of NAD+ modulation on capillary density, perfusion, endothelial nitric oxide (NO) signaling, and mitochondrial reactive oxygen species (ROS) by histology, immunohistochemistry (IHC), qPCR/immunoblot, and functional assays. Build a 3D microfluidic model of diabetic retinal capillary disease from human endothelial/pericyte co-cultures with a retina-on-a-chip model; induce diabetic stress (hyperglycemia + inflammatory cytokines) and apply an NAD+ modulation to map dose–responses for barrier integrity, ROS, NO, and morphology. Analyze publicly available transcriptomic datasets from people with and without diabetic microvascular disease to identify oxidative-stress signatures and candidate circulating biomarkers. Results/Anticipated Results: Expect increasing NAD+ availability to reduce mitochondrial ROS and inflammatory signaling, increase SIRT3/SOD2 activity, improve NO bioavailability, and prevent capillary rarefaction and leak in db/db mice. In the microfluidic model, anticipate a rescue of barrier function, lumen stability, and branching with NAD+, defining an effective dose window. Cross-platform analyses should converge on NAD+-responsive pathways (redox, mitochondrial metabolism, endothelial junctions) and nominate blood biomarkers that correlate with microvascular status. Together, results will provide actionable preclinical evidence for NAD+ modulation as a therapeutic strategy. Discussion/Significance of Impact: By targeting a shared driver, mitochondrial oxidative stress, this work could deliver a mechanism-based therapy and practical biomarkers for diabetic microvascular disease, accelerating translation from bench models to patient-focused trials and improving cardiovascular outcomes.
Kaw et al. (Wed,) studied this question.