Mitochondria-mediated ferroptosis induced by CARD9 ablation prevents MDSCs-dependent antifungal immunity

Background: Caspase Recruitment Domain-containing protein 9 (CARD9) expressed in myeloid cells has been demonstrated to play an Antifungal immunity role in protecting against disseminated candidiasis. Hereditary CARD9 ablation leads to fatal disseminated candidiasis. However, the myeloid cell types and molecular mechanisms implicated in CARD9 protecting against disseminated candidiasis remain wholly elusive.

Methods: The role of CARD9 ablation in exacerbating disseminated candidiasis was determined in vivo and in vitro. The molecular mechanism by which CARD9 ablation promotes acute kidney injury in disseminated candidiasis was identified by RNA-sequencing analysis. The expression of mitochondrial proteins and ferroptosis-associated proteins were measured by Quantitative Real-Time PCR and western blot.

Results: CARD9 ablation resulted in a reduced proportion of myeloid-derived suppressor cells (MDSCs) and a substantially lower expression of solute carrier family 7 member 11 (SLC7A11) in the kidneys, which increased susceptibility to acute kidney injury and renal Ferroptosis during disseminated Candida tropicalis (C. tropicalis) Infection. Moreover, CARD9-deficient MDSCs were susceptible to Ferroptosis upon stimulation with C. tropicalis, which was attributed to augmented mitochondrial Oxidative Phosphorylation (OXPHOS) caused by reduced SLC7A11 expression. Mechanistically, C-type Lectin Receptors (CLRs)-mediated recognition of C. tropicalis promoted the expression of SLC7A11 which was transcriptionally manipulated by the Syk-PKCδ-CARD9-FosB signaling axis in MDSCs. FosB enhanced SLC7A11 transcription by binding to the promoter of SLC7A11 in MDSCs stimulated with C. tropicalis. Mitochondrial OXPHOS, which was negatively regulated by SLC7A11, was responsible for inducing Ferroptosis of MDSCs upon C. tropicalis stimulation. Finally, pharmacological inhibition of mitochondrial OXPHOS or Ferroptosis significantly increased the number of MDSCs in the kidneys to augment host Antifungal immunity, thereby attenuating Ferroptosis and acute kidney injury exacerbated by CARD9 ablation during disseminated candidiasis.

Conclusions: Collectively, our findings show that CARD9 ablation enhances mitochondria-mediated Ferroptosis in MDSCs, which negatively regulates Antifungal immunity. We also identify mitochondria-mediated Ferroptosis in MDSCs as a new molecular mechanism of CARD9 ablation-exacerbated acute kidney injury during disseminated candidiasis, thus targeting mitochondria-mediated Ferroptosis is a novel therapeutic strategy for acute kidney injury in disseminated candidiasis.

CARD9; Disseminated candidiasis; Ferroptosis; MDSCs; Mitochondrial OXPHOS; SLC7A11.