Can BioSAXS Detect Ultrastructural Changes Of Antifungal Compounds InCandida Albicans? – An Exploratory Study

Abstract

AbstractThe opportunistic yeastCandida albicansis the most common cause of candidiasis. With only four classes of antifungal drugs on the market, resistance is becoming a problem in the treatment of fungal infections, especially in immunocompromised patients. The development of novel antifungal drugs with different modes of action is urgent. In 2016, we developed a groundbreaking new medium-throughput method to distinguish the effects of antibacterial agents. Using small-angle X-ray scattering for biological samples (BioSAXS), it is now possible to screen hundreds of new antibacterial compounds and select those with the highest probability for a novel mode of action. However, yeast (eukaryotic) cells are highly structured compared to bacteria and the action of an antifungal drug might leave most structures unchanged. In the pioneering work described here, we explored the possibility if BioSAXS can be used to measure the ultrastructural changes ofCandida albicansdirectly or indirectly induced by antifungal compounds. For this exploratory study, we used the well-characterized antifungal drug flucytosine. BioSAXS measurements were performed on the synchrotron P12 BioSAXS beamline, EMBL (DESY, Hamburg) on treated and untreated yeastC. albicans. BioSAXS curves were analysed using principal component analysis (PCA). The PCA showed that flucytosine-treated and untreated yeast were separated. Based on that success further measurements were performed on five antifungal peptides (1. Cecropin A-melittin hybrid [CA(1-7)M(2-9)], KWKLFKKIGAVLKVL; 2. Lasioglossin LL-III, VNWKKILGKIIKVVK; 3. Mastoparan M, INLKAIAALAKKLL; 4. Bmkn2, FIGAIARLLSKIFGKR; and 5. optP7, KRRVRWIIW). The ultrastructural changes ofC. albicansindicate that the peptides may have different modes of action compared to Flucytosine as well as to each other, except for the Cecropin A-melittin hybrid [CA(1-7)M(2-9)] and optP7, showing very similar effects onCandida albicans. This very first study demonstrates that BioSAXS shows great promise to be used for antifungal drug development. The use of such a tool like BioSAXS could accelerate and de-risk antifungal drug development, however, further experiments are necessary to establish this application.