Identification of a New Class of Antifungals Targeting the Synthesis of Fungal Sphingolipids
Visesato Mor,
Antonella Rella,
Amir M. Farnoud,
Ashutosh Singh,
Mansa Munshi,
Arielle Bryan,
Shamoon Naseem,
James B. Konopka,
Iwao Ojima,
Erika Bullesbach,
Alan Ashbaugh,
Michael J. Linke,
Melanie Cushion,
Margaret Collins,
Hari Krishna Ananthula,
Larry Sallans,
Pankaj B. Desai,
Nathan P. Wiederhold,
Annette W. Fothergill,
William R. Kirkpatrick,
Thomas Patterson,
Lai Hong Wong,
Sunita Sinha,
Guri Giaever,
Corey Nislow,
Patrick Flaherty,
Xuewen Pan,
Gabriele Vargas Cesar,
Patricia de Melo Tavares,
Susana Frases,
Kildare Miranda,
Marcio L. Rodrigues,
Chiara Luberto,
Leonardo Nimrichter,
Maurizio Del Poeta
Affiliations
Visesato Mor
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Antonella Rella
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Amir M. Farnoud
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Ashutosh Singh
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Mansa Munshi
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Arielle Bryan
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Shamoon Naseem
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
James B. Konopka
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
Iwao Ojima
Department of Chemistry and Institute of Chemical Biology and Drug Discovery, Stony Brook University, Stony Brook, New York, USA
Erika Bullesbach
Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA
Alan Ashbaugh
Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA
Michael J. Linke
Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA
Melanie Cushion
Department of Veterans Affairs Medical Center, Cincinnati, Ohio, USA
Margaret Collins
University of Cincinnati College of Medicine, Cincinnati, Ohio, USA
Hari Krishna Ananthula
Department of Pharmaceutical Sciences, University of Cincinnati, Cincinnati, Ohio, USA
Larry Sallans
Department of Chemistry, University of Cincinnati, Cincinnati, Ohio, USA
Pankaj B. Desai
Department of Pharmaceutical Sciences, University of Cincinnati, Cincinnati, Ohio, USA
Nathan P. Wiederhold
Department of Pathology, Fungus Testing Laboratory, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA
Annette W. Fothergill
Department of Pathology, Fungus Testing Laboratory, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA
William R. Kirkpatrick
Division of Infectious Diseases, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA
Thomas Patterson
Division of Infectious Diseases, University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA
Lai Hong Wong
Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada
Sunita Sinha
Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada
Guri Giaever
Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada
Corey Nislow
Department of Pharmaceutical Sciences, University of British Columbia, Vancouver, British Colombia, Canada
Patrick Flaherty
Department of Biomedical Engineering, Worcester Polytechnic Institute, Worcester, Massachusetts, USA
Xuewen Pan
Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, Texas, USA
Gabriele Vargas Cesar
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Patricia de Melo Tavares
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Susana Frases
Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Kildare Miranda
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Marcio L. Rodrigues
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Chiara Luberto
Department of Physiology and Biophysics, Stony Brook University, Stony Brook, New York, USA
Leonardo Nimrichter
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil
Maurizio Del Poeta
Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, New York, USA
ABSTRACT Recent estimates suggest that >300 million people are afflicted by serious fungal infections worldwide. Current antifungal drugs are static and toxic and/or have a narrow spectrum of activity. Thus, there is an urgent need for the development of new antifungal drugs. The fungal sphingolipid glucosylceramide (GlcCer) is critical in promoting virulence of a variety of human-pathogenic fungi. In this study, we screened a synthetic drug library for compounds that target the synthesis of fungal, but not mammalian, GlcCer and found two compounds [N′-(3-bromo-4-hydroxybenzylidene)-2-methylbenzohydrazide (BHBM) and its derivative, 3-bromo-N′-(3-bromo-4-hydroxybenzylidene) benzohydrazide (D0)] that were highly effective in vitro and in vivo against several pathogenic fungi. BHBM and D0 were well tolerated in animals and are highly synergistic or additive to current antifungals. BHBM and D0 significantly affected fungal cell morphology and resulted in the accumulation of intracellular vesicles. Deep-sequencing analysis of drug-resistant mutants revealed that four protein products, encoded by genes APL5, COS111, MKK1, and STE2, which are involved in vesicular transport and cell cycle progression, are targeted by BHBM. IMPORTANCE Fungal infections are a significant cause of morbidity and mortality worldwide. Current antifungal drugs suffer from various drawbacks, including toxicity, drug resistance, and narrow spectrum of activity. In this study, we have demonstrated that pharmaceutical inhibition of fungal glucosylceramide presents a new opportunity to treat cryptococcosis and various other fungal infections. In addition to being effective against pathogenic fungi, the compounds discovered in this study were well tolerated by animals and additive to current antifungals. These findings suggest that these drugs might pave the way for the development of a new class of antifungals.
