ABSTRACT
Aspergillus lentulus was first reported in 2005 as a cryptic species of Aspergillus fumigatus, and since then, its resistance to azole drugs and the high mortality rate of infected individuals have emerged as problems. Although it has been reported that P450 14-α sterol demethylase (Cyp51) is involved in azole resistance in A. lentulus, the specific resistance mechanism has not been elucidated. In this study, we successfully introduced the entire A. fumigatus cyp51A gene into the cyp51A locus in A. lentulus using the CRISPR/Cas9 genome-editing system. The A. lentulus strains harboring A. fumigatus cyp51A showed reduced minimum inhibitory concentrations for itraconazole and voriconazole compared with those of the parent strain. This finding suggests that Cyp51A is involved in azole resistance in A. lentulus and may contribute to the elucidation of the mechanism of resistance to azole drugs via Cyp51A and to the development of new antifungal drugs. In addition, our successful application of the CRISPR/Cas9 system to A. lentulus opens the door to examination of other gene functions in this fungus.
Subject(s)
Azoles , Drug Resistance, Fungal , Antifungal Agents/pharmacology , Aspergillus , Aspergillus fumigatus/genetics , Azoles/pharmacology , CRISPR-Cas Systems , Drug Resistance, Fungal/genetics , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Editing , Humans , Microbial Sensitivity TestsABSTRACT
The authors developed a computer system that automatically produces temporal subtraction images on thin-section thoracic computed tomography based on a 3-dimensional nonlinear geometric warping technique. The automated subtraction system was applied to data from 20 subjects with lung diseases. Nineteen (95%) of 20 results showed good or excellent image quality. The remaining 1 (5%) showed adequate image quality. Our preliminary computer system demonstrates the potential for a clinically useful automated subtraction system.