Caged-hypocrellin mediated antimicrobial photodynamic therapy as a dual-action strategy for fungal clearance and immune response regulation in drug-resistant Candida auris wound infections

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This data is the supplemental material for the manuscript, Caged-hypocrellin mediated antimicrobial photodynamic therapy as a dual-action strategy for fungal clearance and immune response regulation in drug-resistant Candida auris wound infections

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Antifungal susceptibility testing C. auris CBS14918,30 a multidrug-resistant clinical isolate, was used in all experiments. Antifungal susceptibility was assessed following CLSI M27-A3 guidelines.31 Murine skin wound infection model and aPDT BALB/c mice were shaved dorsally, and the skin was disinfected with 75% ethanol followed by sterile saline. Full-thickness wounds were created and inoculated topically with C. auris CBS14918; control mice received sterile saline. Prior to each treatment, the wounds were gently cleansed with saline to remove surface debris, which helped minimize the variability caused by differences in wound conditions such as dryness, exudate, or surface contamination. COP1T-HA (12.5 µg/mL) was applied to the treated wounds, and Phosphate Buffered Saline (PBS) was used as a control. All wounds were then covered with transparent sterile dressings to allow absorption and prevent leakage. Once absorbed, the wounds were illuminated with a 470 nm laser (blue light, 100 mW/cm2) for 30 minutes. The dressing was removed postirradiation. The treatment was repeated daily for three days. Outcomes were assessed at days 3 and 14. Flow cytometry Immune cells from the skin and blood were stained with panels targeting lymphoid and myeloid subsets. Data were acquired on a BD Symphony A5 cytometer and analyzed with FlowJo software. RNA sequencing and analysis Total RNA from wounds and macrophages was sequenced via Illumina NovaSeq. DEGs were identified via DESeq2, and functional enrichment was performed via clusterProfiler.32-35 Enzyme-linked immunosorbent assay (ELISA) and macrophage assays Serum cytokines were measured via ELISA. Primary peritoneal macrophages were cocultured with C. auris and analyzed for transcriptional changes post-aPDT.

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