TY - JOUR AU - Bohner, Flóra AU - Papp, Csaba Gergő AU - Takács, Tamás AU - Varga, Mónika AU - Szekeres, András AU - Nosanchuk, Joshua D. AU - Vágvölgyi, Csaba AU - Tóth, Renáta AU - Gácser, Attila TI - Acquired Triazole Resistance Alters Pathogenicity-Associated Features in Candida auris in an Isolate-Dependent Manner JF - JOURNAL OF FUNGI J2 - J FUNGI VL - 9 PY - 2023 IS - 12 PG - 29 SN - 2309-608X DO - 10.3390/jof9121148 UR - https://m2.mtmt.hu/api/publication/34409072 ID - 34409072 N1 - Department of Microbiology, University of Szeged, Szeged, 6726, Hungary Department of Medicine (Infectious Diseases), Albert Einstein College of Medicine, Bronx, New York, NY 10461, United States Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, NY 10461, United States HCEMM-USZ Fungal Pathogens Research Group, Department of Microbiology, University of Szeged, Szeged, 6726, Hungary HUN-REN-USZ Pathomechanisms of Fungal Infections Research Group, University of Szeged, Szeged, 6726, Hungary Export Date: 30 January 2024 Correspondence Address: Tóth, R.; Department of Microbiology, Hungary; email: renatatth@gmail.com Correspondence Address: Gacser, A.; Department of Microbiology, Hungary; email: gacsera@bio.u-szeged.hu AB - Fluconazole resistance is commonly encountered in Candida auris, and the yeast frequently displays resistance to other standard drugs, which severely limits the number of effective therapeutic agents against this emerging pathogen. In this study, we aimed to investigate the effect of acquired azole resistance on the viability, stress response, and virulence of this species. Fluconazole-, posaconazole-, and voriconazole- resistant strains were generated from two susceptible C. auris clinical isolates (0381, 0387) and compared under various conditions. Several evolved strains became pan-azole-resistant, as well as echinocandin-cross-resistant. While being pan-azole-resistant, the 0381-derived posaconazole-evolved strain colonized brain tissue more efficiently than any other strain, suggesting that fitness cost is not necessarily a consequence of resistance development in C. auris. All 0387-derived evolved strains carried a loss of function mutation (R160S) in BCY1, an inhibitor of the PKA pathway. Sequencing data also revealed that posaconazole treatment can result in ERG3 mutation in C. auris. Despite using the same mechanisms to generate the evolved strains, both genotype and phenotype analysis highlighted that the development of resistance was unique for each strain. Our data suggest that C. auris triazole resistance development is a highly complex process, initiated by several pleiotropic factors. LA - English DB - MTMT ER - TY - JOUR AU - Váradi, Györgyi AU - Kele, Zoltán AU - Czajlik, András AU - Borics, Attila AU - Bende, Gábor AU - Papp, Csaba Gergő AU - Rákhely, Gábor AU - Tóth, Gábor AU - Batta, Gyula AU - Galgóczi, László Norbert TI - Hard nut to crack: Solving the disulfide linkage pattern of the Neosartorya (Aspergillus) fischeri antifungal protein 2 JF - PROTEIN SCIENCE J2 - PROTEIN SCI VL - 32 PY - 2023 IS - 7 PG - 13 SN - 0961-8368 DO - 10.1002/pro.4692 UR - https://m2.mtmt.hu/api/publication/34043893 ID - 34043893 N1 - Department of Medical Chemistry, Albert Szent-Györgyi Medical School, University of Szeged, Szeged, Hungary Department of Organic Chemistry, Faculty of Science and Technology, University of Debrecen, Debrecen, Hungary Department of Biochemistry, Institute of Biochemistry and Molecular Biology, Semmelweis University, Budapest, Hungary Laboratory of Chemical Biology, Institute of Biochemistry, Biological Research Centre, Eötvös Loránd Research Network, Szeged, Hungary Department of Biotechnology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Szeged, Hungary Institute of Biophysics, Biological Research Centre, Eötvös Loránd Research Network, Szeged, Hungary MTA-SZTE Biomimetic Systems Research Group, University of Szeged, Szeged, Hungary Fungal Genomics and Evolution Lab, Institute of Biochemistry, Biological Research Centre, Eötvös Loránd Research Network, Szeged, Hungary Export Date: 11 August 2023 CODEN: PRCIE Correspondence Address: Váradi, G.; Department of Medical Chemistry, Dóm tér 8, Hungary; email: varadi.gyorgyi@med.u-szeged.hu Correspondence Address: Galgóczy, L.; Department of Biotechnology, Közép fasor 52, Hungary; email: galgoczi@bio.u-szeged.hu Chemicals/CAS: disulfide, 16734-12-6; Antifungal Agents; Disulfides AB - As a consequence of the fast resistance spreading, a limited number of drugs are available to treat fungal infections. Therefore, there is an urgent need to develop new antifungal treatment strategies. The features of a disulfide bond-stabilized antifungal protein, NFAP2 secreted by the mold Neosartorya (Aspergillus) fischeri render it to be a promising template for future protein-based antifungal drug design, which requires knowledge about the native disulfide linkage pattern as it is one of the prerequisites for biological activity. However, in the lack of tryptic and chymotryptic proteolytic sites in the ACNCPNNCK sequence, the determination of the disulfide linkage pattern of NFAP2 is not easy with traditional mass spectrometry-based methods. According to in silico predictions working with a preliminary nuclear magnetic resonance (NMR) solution structure, two disulfide isomers of NFAP2 (abbacc and abbcac) were possible. Both were chemically synthesized; and comparative reversed-phase high-performance liquid chromatography, electronic circular dichroism and NMR spectroscopy analyses, and antifungal susceptibility and efficacy tests indicated that the abbcac is the native pattern. This knowledge allowed rational modification of NAFP2 to improve the antifungal efficacy and spectrum through the modulation of the evolutionarily conserved gamma-core region, which is responsible for the activity of several antimicrobial peptides. Disruption of the steric structure of NFAP2 upon gamma-core modification led to the conclusions that this motif may affect the formation of the biologically active three-dimensional structure, and that the gamma-core modulation is not an efficient tool to improve the antifungal efficacy or to change the antifungal spectrum of NFAP2. LA - English DB - MTMT ER - TY - CONF AU - Bohner, Flóra AU - Papp, Csaba Gergő AU - Takács, Tamás AU - Szekeres, András AU - Varga, Mónika AU - Vágvölgyi, Csaba AU - Tóth, Renáta AU - Gácser, Attila TI - ACQUISITION OF ANTIFUNGAL RESISTANCE INFLUENCES VIRULENCE ATTRIBUTES OF CANDIDA AURIS MICROEVOLVED STRAINS IN MOUSE SYSTEMIC FUNGAL INFECTION MODEL T2 - A Magyar Mikrobiológiai Társaság 2022. évi Nagygyűlése és a XV. Fermentációs Kollokvium PY - 2022 SP - 11 UR - https://m2.mtmt.hu/api/publication/33576180 ID - 33576180 LA - English DB - MTMT ER - TY - CHAP AU - Tóth, Liliána AU - Papp, Rebeka AU - Váradi, Györgyi AU - Tóth, K Gábor AU - Papp, Csaba Gergő AU - Rákhely, Gábor AU - Poór, Péter AU - Galgóczi, László Norbert ED - Bánfalvi, Zsófia ED - Gócza, Elen ED - Olasz, Ferenc ED - Pál, Magda ED - Posta, Katalin ED - Várallyay, Éva TI - Antifungal effect of a novel defensin from Solanum lycopersicum L., and it's γ-core peptide derivatives T2 - „FIBOK 2022” : Fiatal Biotechnológusok V. Országos Konferenciája PB - MATE Genetika és Biotechnológia Intézet CY - Gödöllő SN - 9789632699998 PY - 2022 SP - 33 EP - 33 PG - 1 UR - https://m2.mtmt.hu/api/publication/33255670 ID - 33255670 LA - English DB - MTMT ER - TY - CONF AU - Bende, Gábor AU - Zsindely, Nóra AU - Bodai, László AU - Krisztián, Laczi AU - Papp, Csaba Gergő AU - Attila, Farkas AU - Zsolt, Kristóffy AU - Petra, Királdi AU - Gábor, Rákhely AU - Galgóczi, László Norbert TI - Characterization of NFAP2 resistance-evolved Candida albicans strains T2 - A Magyar Mikrobiológiai Társaság 2022. évi Nagygyűlése és a XV. Fermentációs Kollokvium PY - 2022 SP - 8 EP - 8 PG - 1 UR - https://m2.mtmt.hu/api/publication/33252438 ID - 33252438 LA - English DB - MTMT ER - TY - CHAP AU - Tóth, Liliána AU - Papp, Rebeka AU - Papp, Csaba Gergő AU - Váradi, Györgyi AU - Tóth, Gábor AU - Gábor, Rákhely AU - Poór, Péter AU - Galgóczi, László Norbert TI - Antifungal activity of peptides designed on the γ–core of tomato plant defensins T2 - FEMS Conference on Microbiology in association with Serbian Society of Microbiology PB - Serbian Society of Microbiology CY - Belgrád SN - 9788691489786 PY - 2022 SP - 837 EP - 838 PG - 2 UR - https://m2.mtmt.hu/api/publication/33250266 ID - 33250266 LA - English DB - MTMT ER - TY - JOUR AU - Kakar, Anant AU - Sastre-Velasquez, Luis Enrique AU - Hess, Michael AU - Galgóczi, László Norbert AU - Papp, Csaba Gergő AU - Holzknecht, Jeanett AU - Romanelli, Alessandra AU - Váradi, Györgyi AU - Malanovic, Nermina AU - Marx, Florentine TI - The Membrane Activity of the Amphibian Temporin B Peptide Analog TB_KKG6K Sheds Light on the Mechanism That Kills Candida albicans JF - mSPHERE J2 - MSPHERE VL - 7 PY - 2022 IS - 5 PG - 20 SN - 2379-5042 DO - 10.1128/msphere.00290-22 UR - https://m2.mtmt.hu/api/publication/33069296 ID - 33069296 N1 - Funding Agency and Grant Number: Austrian Science Fund FWF (HOROS Doctoral Program) [W1253 DK HOROS]; Hungarian National Research, Development and Innovation Office (NKFIH) [FK 134343] Funding text: We thank W. Salvenmoser for helpful discussion, and D. Bratschun-Khan, K. Gutleben, and B. Witting for technical assistance. The study was funded by the Austrian Science Fund FWF (HOROS Doctoral Program, W1253 DK HOROS) to F.M. and L.G. was financed by the Hungarian National Research, Development and Innovation Office (NKFIH, FK 134343 project). We have no conflict of interest to declare. AB - Fungal infections with the opportunistic human pathogen C. albicans are associated with high mortality rates in immunocompromised patients. This is partly due to the yeast's ability to rapidly develop resistance toward currently available antifungals. Temporin B (TB) is a 13-amino-acid-long, cationic peptide secreted by the granular glands of the European frog Rana temporaria. We recently showed that the modified TB peptide analog TB_KKG6K rapidly killed planktonic and sessile Candida albicans at low micromolar concentrations and was neither hemolytic nor cytotoxic to mammalian cells in vitro. The present study aimed to shed light into its mechanism of action, with a focus on its fungal cell membrane activity. We utilized different fluorescent dyes to prove that it rapidly induces membrane depolarization and permeabilization. Studies on model membrane systems revealed that the TB analog undergoes hydrophobic and electrostatic membrane interactions, showing a preference for anionic lipids, and identified phosphatidylinositol and cardiolipin as possible peptide targets. Fluorescence microscopy using fluorescein isothiocyanate-labeled TB_KKG6K in the presence of the lipophilic dye FM4-64 indicated that the peptide compromises membrane integrity and rapidly enters C. albicans cells in an energy-independent manner. Peptide-treated cells analyzed by cryo-based electron microscopy exhibited no signs of cell lysis; however, subcellular structures had disintegrated, suggesting that intracellular activity may form part of the killing mechanism of the peptide. Taken together, this study proved that TB_KKG6K compromises C. albicans membrane function, which explains the previously observed rapid, fungicidal mode of action and supports its great potential as a future anti-Candida therapeutic. IMPORTANCE Fungal infections with the opportunistic human pathogen C. albicans are associated with high mortality rates in immunocompromised patients. This is partly due to the yeast's ability to rapidly develop resistance toward currently available antifungals. Small, cationic, membrane-active peptides are promising compounds to fight against resistance development, as many of them effectuate rapid fungal cell death. This fast killing is believed to hamper the development of resistance, as the fungi do not have sufficient time to adapt to the antifungal compound. We previously reported that the synthetic variant of the amphibian TB peptide, TB_KKG6K, rapidly kills C. albicans. In the current study, the mechanism of action of the TB analog was investigated. We show that this TB analog is membrane-active and impairs cell membrane function, highlighting its potential to be developed as an attractive alternative anti-C. albicans therapeutic that may hinder the development of resistance. LA - English DB - MTMT ER - TY - JOUR AU - Igaz, Nóra AU - Bélteky, Péter AU - Kovács, Dávid AU - Papp, Csaba Gergő AU - Rónavári, Andrea AU - Szabó, Diána AU - Gácser, Attila AU - Kónya, Zoltán AU - Csontné Kiricsi, Mónika TI - Functionalized Mesoporous Silica Nanoparticles for Drug-Delivery to Multidrug-Resistant Cancer Cells JF - INTERNATIONAL JOURNAL OF NANOMEDICINE J2 - INT J NANOMED VL - 17 PY - 2022 SP - 3079 EP - 3096 PG - 18 SN - 1176-9114 DO - 10.2147/IJN.S363952 UR - https://m2.mtmt.hu/api/publication/33003489 ID - 33003489 N1 - Department of Biochemistry and Molecular Biology, University of Szeged, Szeged, Hungary Department of Applied and Environmental Chemistry, University of Szeged, Szeged, Hungary Institut de Pharmacologie Moléculaire et Cellulaire, Université Côte d’Azur, Inserm, CNRS, Valbonne, France HCEMM-USZ Fungal Pathogens Research Group, Department of Microbiology, University of Szeged, Szeged, Hungary Department of Oto-Rhino-Laryngology and Head & Neck Surgery, Szeged, Hungary Eötvös Loránd Research Network, Reaction Kinetics and Surface Chemistry Research Group, Szeged, Hungary Cited By :8 Export Date: 27 February 2024 Correspondence Address: Kónya, Z.; Department of Applied and Environmental Chemistry, Rerrich square 1, Hungary; email: konya@chem.u-szeged.hu Correspondence Address: Kiricsi, M.; Department of Biochemistry and Molecular Biology, Közép fasor 52, Hungary; email: kiricsim@bio.u-szeged.hu AB - Background: Multidrug resistance is a common reason behind the failure of chemotherapy. Even if the therapy is effective, serious adverse effects might develop due to the low specificity and selectivity of antineoplastic agents. Mesoporous silica nanoparticles (MSNs) are promising materials for tumor-targeting and drug-delivery due to their small size, relatively inert nature, and extremely large specific surfaces that can be functionalized by therapeutic and targeting entities. We aimed to create a fluorescently labeled MSN-based drug-delivery system and investigate their internalization and drug-releasing capability in drug-sensitive MCF-7 and P-glycoprotein-overexpressing multidrug-resistant MCF-7 KCR cancer cells.Methods and Results: To track the uptake and subcellular distribution of MSNs, particles with covalently coupled red fluorescent Rhodamine B (RhoB) were produced (RhoB@MSNs). Both MCF-7 and MCF-7 KCR cells accumulated a significant amount of RhoB@MSNs. The intracellular RhoB@MSN concentrations did not differ between sensitive and multidrug-resistant cells and were kept at the same level even after cessation of RhoB@MSN exposure. Although most RhoB@MSNs resided in the cytoplasm, significantly more RhoB@MSNs co-localized with lysosomes in multidrug-resistant cells compared to sensitive counterparts. To examine the drug-delivery capability of these particles, RhoB@Rho123@MSNs were established, where RhoB-functionalized nanoparticles carried green fluorescent Rhodamine 123 (Rho123) -a P-glycoprotein substrate - as cargo within mesopores. Significantly higher Rho123 fluorescence intensity was detected in RhoB@Rho123@MSN-treated multidrug-resistant cells than in free Rho123-exposed counterparts. The exceptional drug-delivery potential of MSNs was further verified using Mitomycin C (MMC)-loaded RhoB@MSNs (RhoB@MMC@MSNs). Exposures to RhoB@MMC@MSNs significantly decreased the viability not only of drug-sensitive but of multidrug-resistant cells and the elimination of MDR cells was significantly more robust than upon free MMC treatments.Conclusion: The efficient delivery of Rho123 and MMC to multidrug-resistant cells via MSNs, the amplified and presumably prolonged intracellular drug concentration, and the consequently enhanced cytotoxic effects envision the enormous potential of MSNs to defeat multidrug-resistant cancer. LA - English DB - MTMT ER - TY - JOUR AU - Bohner, Flóra AU - Papp, Csaba Gergő AU - Gácser, Attila TI - The effect of antifungal resistance development on the virulence of Candida species JF - FEMS YEAST RESEARCH J2 - FEMS YEAST RES VL - 22 PY - 2022 IS - 1 PG - 10 SN - 1567-1356 DO - 10.1093/femsyr/foac019 UR - https://m2.mtmt.hu/api/publication/32763732 ID - 32763732 N1 - HCEMM-USZ Fungal Pathogens Research Group, Department of Microbiology, Faculty of Science and Informatics, University of Szeged, Szeged, H-6726, Hungary MTA-SZTE 'Lendület', Mycobiome Research Group, University of Szeged, Szeged, H-6726, Hungary Cited By :4 Export Date: 25 January 2023 CODEN: FYREA Correspondence Address: Gácser, A.; University of Szeged, Közép fasor 52., Hungary; email: gacsera@bio.u-szeged.hu Chemicals/CAS: amphotericin B, 1397-89-3, 30652-87-0; echinocandin, 80619-41-6; Antifungal Agents; Azoles; Echinocandins; Polyenes LA - English DB - MTMT ER - TY - JOUR AU - Tóth, Liliána AU - Poór, Péter AU - Ördög, Attila AU - Váradi, Györgyi AU - Farkas, Attila AU - Papp, Csaba Gergő AU - Bende, Gábor AU - Tóth, Gábor AU - Rákhely, Gábor AU - Marx, Florentine AU - Galgóczi, László Norbert TI - The combination of Neosartorya (Aspergillus) fischeri antifungal proteins with rationally designed γ-core peptide derivatives is effective for plant and crop protection JF - BIOCONTROL J2 - BIOCONTROL VL - 67 PY - 2022 IS - 2 SP - 249 EP - 262 PG - 14 SN - 1386-6141 DO - 10.1007/s10526-022-10132-y UR - https://m2.mtmt.hu/api/publication/32655436 ID - 32655436 N1 - Export Date: 23 June 2022 CODEN: BOCOF AB - Plant pathogenic fungi are responsible for enormous crop losses worldwide. Overcoming this problem is challenging as these fungi can be highly resistant to approved chemical fungicides. There is thus a need to develop and introduce fundamentally new plant and crop protection strategies for sustainable agricultural production. Highly stable extracellular antifungal proteins (AFPs) and their rationally designed peptide derivatives (PDs) constitute feasible options to meet this challenge. In the present study, their potential for topical application to protect plants and crops as combinatorial biofungicides is supported by the investigation of two Neosartorya (Aspergillus) fischeri AFPs (NFAP and NFAP2) and their γ-core PDs. Previously, the biofungicidal potential of NFAP, its rationally designed γ-core PD (γNFAP-opt), and NFAP2 was reported. Susceptibility tests in the present study extended the in vitro antifungal spectrum of NFAP2 and its γ-core PD (γNFAP2-opt) to Botrytis, Cladosporium, and Fusarium spp. Besides, in vitro additive or indifferent interactions, and synergism were observed when NFAP or NFAP2 was applied in combination with γNFAP-opt. Except for γNFAP2-opt, the investigated proteins and peptides did not show any toxicity to tomato plant leaves. The application of NFAP in combination with γNFAP-opt effectively inhibited conidial germination, biofilm formation, and hyphal extension of the necrotrophic mold Botrytis cinerea on tomato plant leaves. However, the same combination only partially impeded the B. cinerea-mediated decay of tomato fruits, but mitigated the symptoms. Our results highlight the feasibility of using the combination of AFP and PD as biofungicide for the fungal infection control in plants and crops. LA - English DB - MTMT ER -