The Aspergillus fumigatus C2-Domain Protein SppA is required for septal integrity and alters susceptibility to echinocandins and neutrophil killing during infection

Aspergillus fumigatus is a major opportunistic fungal pathogen whose ability to maintain hyphal integrity and withstand host defenses is critical for virulence. Septal pores, which connect hyphal compartments, are dynamically regulated to preserve cellular integrity under stress, yet the molecular components governing this process remain incompletely defined. Here, we identify and characterize a septal pore-associated protein, SppA, and demonstrate its essential role in maintaining septal integrity in A. fumigatus. We show that expression of SppA is positively regulated by the transcription factor ZfpA and is induced in response to the cell wall-targeting antifungal caspofungin. Deletion of sppA resulted in defective septal organization and increased susceptibility to hyphal damage. The mutant exhibited heightened sensitivity to cell wall-targeting antifungal agents, indicating a role in cell wall stress tolerance. In a zebrafish model of invasive aspergillosis, loss of SppA significantly attenuated virulence which was abrogated in neutrophil-deficient zebrafish. Further, the mutant strain displayed increased susceptibility to killing by primary human neutrophils, suggesting that proper septal pore formation contributes to fungal survival during host immune attack. Together, our findings establish SppA as a critical determinant of septal integrity, antifungal tolerance, and pathogenicity in A. fumigatus, and position it as part of a ZfpA-regulated, caspofungin-responsive pathway that supports fungal survival during stress and infection. Author SummaryAspergillus fumigatus is a common environmental mold that can cause life-threatening infections in people with weakened immune systems. For successful invasion of host tissue, the fungus requires the ability to protection sections of its hyphae from cell wall targeting antifungals and host immune cell attack by closing septal (cross wall) pores distributed throughout hyphal strands. We have identified an A. fumigatus protein, SppA, required for proper septal pore closure. Loss of SppA reduces the ability of hyphae to withstand treatment with antifungals and the ability of A. fumigatus to cause disease in a zebrafish infection model. The SppA mutant was particularly susceptible to killing by neutrophils, key immune cells that help control fungal infections. Our findings reveal an important mechanism that helps A. fumigatus survive environmental and host-imposed stresses and highlight septal pore regulation as a potential target for future antifungal strategies.