Cholesterol import by Aspergillus fumigatus and its influence on antifungal potency of sterol biosynthesis inhibitors.
High mortality rates from invasive aspergillosis in immunocompromised patients are prompting research toward improved antifungal therapy and better understanding of fungal physiology. Herein we show that Aspergillus fumigatus, the major pathogen in aspergillosis, imports exogenous cholesterol under aerobic conditions and thus compromises the antifungal potency of sterol biosynthesis inhibitors. Adding serum to RPMI medium led to enhanced growth of A. fumigatus and extensive import of cholesterol, most of which was stored as ester. Growth enhancement and sterol import also occurred when the medium was supplemented with purified cholesterol instead of serum. Cells cultured in RPMI medium with the sterol biosynthesis inhibitors itraconazole or voriconazole showed retarded growth, a dose-dependent decrease in ergosterol levels, and accumulation of aberrant sterol intermediates. Adding serum or cholesterol to the medium partially rescued the cells from the drug-induced growth inhibition. We conclude that cholesterol import attenuates the potency of sterol biosynthesis inhibitors, perhaps in part by providing a substitute for membrane ergosterol. Our findings establish significant differences in sterol homeostasis between filamentous fungi and yeast. These differences indicate the potential value of screening aspergillosis antifungal agents in serum or other cholesterol-containing medium. Our results also suggest an explanation for the antagonism between itraconazole and amphotericin B, the potential use of Aspergillus as a model for sterol trafficking, and new insights for antifungal drug development.
Xiong
Quanbo
Q
Department of Biochemistry and Cell Biology, Rice University, 6100 Main St., Houston, TX 77005, USA.
Hassan
Saad A
SA
Wilson
William K
WK
Han
Xiang Y
XY
May
Gregory S
GS
Tarrand
Jeffrey J
JJ
Matsuda
Seiichi P T
SP
D000935:Antifungal Agents / Q000494:pharmacology*; D001232:Aspergillus fumigatus / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D001393:Azoles / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry; D002784:Cholesterol / Q000378:metabolism*; D017077:Culture Media, Conditioned; D004875:Ergosterol / Q000096:biosynthesis; D006801:Humans; D017964:Itraconazole / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D010101:Oxygen Consumption; D011743:Pyrimidines / Q000494:pharmacology; D013172:Spores, Fungal / Q000187:drug effects / Q000254:growth & development; D013261:Sterols / Q000037:antagonists & inhibitors* / Q000096:biosynthesis*; D014230:Triazoles / Q000494:pharmacology; D065819:Voriconazole
D000935:Antifungal Agents; D001393:Azoles; D017077:Culture Media, Conditioned; D011743:Pyrimidines; D013261:Sterols; D014230:Triazoles; D017964:Itraconazole; D002784:Cholesterol; D065819:Voriconazole; D004875:Ergosterol