Metabolic regulation of innate immunity-dependent wound healing in situ

The focus of this application is to understand the integrated metabolic regulation of wound healing. Productive tissue repair is a fundamental requirement for an organism to maintain tissue homeostasis and organ function. Dysregulated wound healing is relevant to various human diseases, including heart disease, autoimmune diseases, chronic wounds, cancer, and infection. Tissue repair requires the coordinated action of multiple cell types, including immune and stromal cells, through several phases of wound healing that take days to months. Intracellular metabolism is a central regulator of cellular effector functions and has become a therapeutic target to modulate disease progression and patient outcome. Despite the progress in understanding the signaling networks that regulate wound healing, the metabolic regulation of wound healing in vivo remains poorly understood. Key gaps include understanding how the intracellular metabolism of individual cells is regulated to influence effector functions and how metabolic crosstalk between different cell types is coordinated for productive wound healing. We use zebrafish (Danio rerio) as our in vivo model of inflammation and tissue repair. The optical transparency, ease of genetic manipulation, and high similarity to the human immune system and genome make zebrafish an ideal model for dissecting multi-cellular interactions during tissue repair in situ. We have shown that fluorescence lifetime imaging microscopy (FLIM) of endogenous metabolic coenzymes, nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) are sensitive to dynamic changes in the intracellular metabolism of macrophages during wound responses in live zebrafish larvae. We will continue implementing FLIM modalities and analytical tools to enable the spatiotemporal analysis of intracellular metabolism in native interstitial spaces in a living animal. We found that mitochondrial ROS contributes to macrophage metabolism during the early pro-inflammatory phase of wound response. We will employ single cell-based imaging, including FLIM and fluorescent metabolic sensors using standard confocal microscopy, and spatial metabolomics to dissect the role of canonical and non-canonical pathways in mitochondrial function to regulate innate immunity and interaction with epithelial cells during tissue repair. Our studies will contribute to a better understanding of the metabolic regulation of tissue repair and may provide new targets to treat human disease. Project Number: 1R35GM162265-01 | Fiscal Year: 2026 | NIH Institute/Center: National Institute of General Medical Sciences (NIGMS) | Principal Investigator: Veronika Miskolci | Institution: RUTGERS BIOMEDICAL AND HEALTH SCIENCES, Newark, NJ | Award Amount: $321,304 | Activity Code: R35 | Study Section: Special Emphasis Panel[ZRG1 CDB-N (55)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11271679