Molecule Information

General Information of the Molecule (ID: Mol01088)

• Name: Sterol 14-alpha demethylase cyp51A (CYP51A) ,Aspergillus fumigatus
• Synonyms: Cytochrome P450 monooxygenase 51A; Ergosterol biosynthesis protein 11A; erg11A; AFUA_4G06890
• Molecule Type: Protein
• Gene Name: cyp51A
• Gene ID: 3509526
• Sequence:
  MVPMLWLTAYMAVAVLTAILLNVVYQLFFRLWNRTEPPMVFHWVPYLGSTISYGIDPYKF
  FFACREKYGDIFTFILLGQKTTVYLGVQGNEFILNGKLKDVNAEEVYSPLTTPVFGSDVV
  YDCPNSKLMEQKKFIKYGLTQSALESHVPLIEKEVLDYLRDSPNFQGSSGRVDISAAMAE
  ITIFTAARALQGQEVRSKLTAEFADLYHDLDKGFTPINFMLPWAPLPHNKKRDAAHARMR
  SIYVDIITQRRLDGEKDSQKSDMIWNLMNCTYKNGQQVPDKEIAHMMITLLMAGQHSSSS
  ISAWIMLRLASQPKVLEELYQEQLANLGPAGPDGSLPPLQYKDLDKLPFHQHVIRETLRI
  HSSIHSIMRKVKSPLPVPGTPYMIPPGRVLLASPGVTALSDEHFPNAGCWDPHRWENQAT
  KEQENDKVVDYGYGAVSKGTSSPYLPFGAGRHRCIGEKFAYVNLGVILATIVRHLRLFNV
  DGKKGVPETDYSSLFSGPMKPSIIGWEKRSKNTSK
• Function: Sterol 14-alpha demethylase; part of the third module of ergosterol biosynthesis pathway that includes the late steps of the pathway (Probable). The third module or late pathway involves the ergosterol synthesis itself through consecutive reactions that mainly occur in the endoplasmic reticulum (ER) membrane. Firstly, the squalene synthase erg9 catalyzes the condensation of 2 farnesyl pyrophosphate moieties to form squalene, which is the precursor of all steroids. Squalene synthase is crucial for balancing the incorporation of farnesyl diphosphate (FPP) into sterol and nonsterol isoprene synthesis. Secondly, squalene is converted into lanosterol by the consecutive action of the squalene epoxidase erg1 and the lanosterol synthase erg7. Then, the delta(24)-sterol C-methyltransferase erg6 methylates lanosterol at C-24 to produce eburicol. Eburicol is the substrate of the sterol 14-alpha demethylase encoded by cyp51A and cyp51B, to yield 4,4,24-trimethyl ergosta-8,14,24(28)-trienol. The C-14 reductase erg24 then reduces the C14=C15 double bond which leads to 4,4-dimethylfecosterol. A sequence of further demethylations at C-4, involving the C-4 demethylation complex containing the C-4 methylsterol oxidases erg25A or erg25B, the sterol-4-alpha-carboxylate 3-dehydrogenase erg26 and the 3-keto-steroid reductase erg27, leads to the production of fecosterol via 4-methylfecosterol. The C-8 sterol isomerase erg2 then catalyzes the reaction which results in unsaturation at C-7 in the B ring of sterols and thus converts fecosterol to episterol. The sterol-C5-desaturase erg3B then catalyzes the introduction of a C-5 double bond in the B ring to produce 5-dehydroepisterol. The 2 other sterol-C5-desaturases, erg3A and erg3C, seem to be less important in ergosterol biosynthesis. The C-22 sterol desaturase erg5 further converts 5-dehydroepisterol into ergosta-5,7,22,24(28)-tetraen-3beta-ol by forming the C-22(23) double bond in the sterol side chain. Finally, ergosta-5,7,22,24(28)-tetraen-3beta-ol is substrate of the C-24(28) sterol reductases erg4A and erg4B to produce ergosterol. Possible alternative sterol biosynthetic pathways might exist from fecosterol to ergosterol, depending on the activities of the erg3 isoforms.
• Uniprot ID: CP51A_ASPFU

Kingdom: Fungi
Phylum: Ascomycota
Class: Eurotiomycetes
Order: Eurotiales
Family: Aspergillaceae
Genus: Aspergillus
Species: Aspergillus fumigatus

Type(s) of Resistant Mechanism of This Molecule

  ADTT: Aberration of the Drug's Therapeutic Target

Drug Resistance Data Categorized by Drug

Approved Drug(s) 3 drug(s) in total

Itraconazole

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54E
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: Itraconazole resistance has been tightly linked to cyp51A mutations in the codon for Gly54, resulting in five different amino substitutions (G54k, G54V, G54R, G54E, and G54W).

Disease Class: Aspergillus fumigatus infection [2]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.F46Y+p.M172V+p.N248T+p.D255E
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST broth dilution method assay
• Mechanism Description: Interestingly, the F46Y/M172V/N248T/D255E/E427k mutation, which has been reported to be associated with azole resistance (37), was detected in one clinical isolate from Shanghai and in one environmental isolate from Xinjiang, respectively.

Disease Class: Aspergillosis [3]

• Resistant Disease: Aspergillosis [ICD-11: 1F20.0]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.I266N
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: Amino acid changes in CYP51a may contribute to Aspergillus fumigatus emerging itraconazole resistance.

Disease Class: Aspergillus fumigatus infection [4]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54E
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: A Point Mutation in the 14alpha-Sterol Demethylase Gene cyp51A Contributes to Itraconazole Resistance in Aspergillus fumigatus.

Disease Class: Aspergillus fumigatus infection [5]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54E+p.I266N
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: Amino acid changes in CYP51a may contribute to Aspergillus fumigatus emerging itraconazole resistance.

Disease Class: Aspergillus fumigatus infection [6]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.M220L
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: PCR amplification and sequence analysis
• Experiment for Drug Resistance: NCCLS method M-27A with broth macrodilution techniques assay
• Mechanism Description: Five clinical isolates of Aspergillus fumigatus that exhibited similar patterns of reduced susceptibility to itraconazole and other triazole drugs were analyzed. Sequence analysis of genes (cyp51A and cyp51B) encoding the 14alpha-sterol demethylases revealed that all five strains harbored mutations in cyp51A resulting in the replacement of methionine at residue 220 by valine, lysine, or threonine. When the mutated cyp51A genes were introduced into an A. fumigatus wild-type strain, the transformants exhibited reduced susceptibility to all triazole agents, confirming that the mutations were responsible for the resistance phenotype.

Disease Class: Aspergillus fumigatus infection [6]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.M220V
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: PCR amplification and sequence analysis
• Experiment for Drug Resistance: NCCLS method M-27A with broth macrodilution techniques assay
• Mechanism Description: Five clinical isolates of Aspergillus fumigatus that exhibited similar patterns of reduced susceptibility to itraconazole and other triazole drugs were analyzed. Sequence analysis of genes (cyp51A and cyp51B) encoding the 14alpha-sterol demethylases revealed that all five strains harbored mutations in cyp51A resulting in the replacement of methionine at residue 220 by valine, lysine, or threonine. When the mutated cyp51A genes were introduced into an A. fumigatus wild-type strain, the transformants exhibited reduced susceptibility to all triazole agents, confirming that the mutations were responsible for the resistance phenotype.

Disease Class: Aspergillus fumigatus infection [6]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.M220T
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: PCR amplification and sequence analysis
• Experiment for Drug Resistance: NCCLS method M-27A with broth macrodilution techniques assay
• Mechanism Description: Five clinical isolates of Aspergillus fumigatus that exhibited similar patterns of reduced susceptibility to itraconazole and other triazole drugs were analyzed. Sequence analysis of genes (cyp51A and cyp51B) encoding the 14alpha-sterol demethylases revealed that all five strains harbored mutations in cyp51A resulting in the replacement of methionine at residue 220 by valine, lysine, or threonine. When the mutated cyp51A genes were introduced into an A. fumigatus wild-type strain, the transformants exhibited reduced susceptibility to all triazole agents, confirming that the mutations were responsible for the resistance phenotype.

Disease Class: Aspergillus fumigatus infection [7]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.F219S
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Multivariate analysis of overall survival or disease-free survival assay
• Mechanism Description: Each single mutationad a measurable effect on the affinity of the target enzyme for specific azole derivatives.

Disease Class: Aspergillus fumigatus infection [7]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G138S
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Multivariate analysis of overall survival or disease-free survival assay
• Mechanism Description: Each single mutationad a measurable effect on the affinity of the target enzyme for specific azole derivatives.

Disease Class: Aspergillus fumigatus infection [7]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G138C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Multivariate analysis of overall survival or disease-free survival assay
• Mechanism Description: Each single mutationad a measurable effect on the affinity of the target enzyme for specific azole derivatives.

Disease Class: Aspergillus fumigatus infection [8]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Itraconazole
• Molecule Alteration: Tandem repeat; TR53 (GAATCACGCGGTCCGATGTGTGCTGAGCCGAATGAAAGTTGTCTAATGTCTAGAATCACGCGGTCCGATGTGTGCTGAGCCGAATGAAAGTTGTCTAATGTCTA)
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: RT-PCR
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: The azole-resistant A. fumigatus strains were detected tandem repeats (TRs) in the promoter region.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54V
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: Itraconazole resistance has been tightly linked to cyp51A mutations in the codon for Gly54, resulting in five different amino substitutions (G54k, G54V, G54R, G54E, and G54W).

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54K
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: The G54k amino acid change conferred cross-resistance to both itraconazole and posaconazole (26). The replacement of the wild-type chromosomal cyp51A allele by mutant allele bearing the G161A nucleotide change in codon 54 led to the acquisition of resistance to itraconazole de novo.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G161A
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: The G54k amino acid change conferred cross-resistance to both itraconazole and posaconazole (26). The replacement of the wild-type chromosomal cyp51A allele by mutant allele bearing the G161A nucleotide change in codon 54 led to the acquisition of resistance to itraconazole de novo.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54R
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: The G54k amino acid change conferred cross-resistance to both itraconazole and posaconazole (26). The replacement of the wild-type chromosomal cyp51A allele by mutant allele bearing the G161A nucleotide change in codon 54 led to the acquisition of resistance to itraconazole de novo.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54W
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: Itraconazole resistance has been tightly linked to cyp51A mutations in the codon for Gly54, resulting in five different amino substitutions (G54k, G54V, G54R, G54E, and G54W).

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.P216L
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.H147Y
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G434C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.Y431C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [10]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.Y121F+p.T289A+p.G448S+p.M172I
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain TR463; 746128
• Experiment for Molecule Alteration: PCR analysis
• Mechanism Description: In addition, to compare the susceptibility of TR463 with those of TR34 and TR46, the high resistance of TR463/Y121F/M172I/T289A/G448S was confirmed by MIC testing, displaying a pan-triazole-resistant phenotype to posaconazole, itraconazole, and voriconazole, indicating no in vitro activity of itraconazole and voriconazole (MIC, >16 mg/liter).

Disease Class: Invasive aspergillosis [11]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G138C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST broth dilution method assay
• Mechanism Description: Three different cyp51A mutations were found (G138C, Y431C, and G434C), of which the first two were demonstrated by heterologous expression in a hypersusceptible Saccharomyces cerevisiae strain to be at least partly responsible for elevated MICs.

Disease Class: Invasive aspergillosis [11]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.Y431C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST broth dilution method assay
• Mechanism Description: Three different cyp51A mutations were found (G138C, Y431C, and G434C), of which the first two were demonstrated by heterologous expression in a hypersusceptible Saccharomyces cerevisiae strain to be at least partly responsible for elevated MICs.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.G54W
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: RT-PCR
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: A. fumigatus is closely linked to amino acid substitutions in Cyp51A that replace Gly54

Disease Class: Invasive aspergillosis [12]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.L98H
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: We have determined that a base change causing an amino acid substitution in Cyp51A (L98H) in combination with the duplication in tandem of a 34-bp sequence in the cyp51A promoter, which is responsible for the increased level of cyp51A gene expression, accounted for the resistant phenotype.

Disease Class: Pulmonary aspergillosis [13]

• Resistant Disease: Pulmonary aspergillosis [ICD-11: 1F20.7]
• Resistant Drug: Itraconazole
• Molecule Alteration: Missense mutation; p.M217I
• Experimental Note: Identified from the Human Clinical Data
• Mechanism Description: Itraconazole acts by inhibiting the fungal cytochrome P-450 dependent enzyme lanosterol 14-alpha-demethylase.The development of itraconazole resistance in A. terreus which may be associated with M217I Cyp51A mutation.

Posaconazole

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Disease Class: Aspergillus fumigatus infection [2]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.F46Y+p.M172V+p.N248T+p.D255E
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST broth dilution method assay
• Mechanism Description: Interestingly, the F46Y/M172V/N248T/D255E/E427k mutation, which has been reported to be associated with azole resistance (37), was detected in one clinical isolate from Shanghai and in one environmental isolate from Xinjiang, respectively.

Disease Class: Aspergillus fumigatus infection [14]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.G54W
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: In the four isolates exhibiting moderate levels of POS resistance glycine 54 was mutated to either glutamate or arginine. In the mutant with a high level of POS resistance glycine 54 was mutated to tryptophan.

Disease Class: Aspergillus fumigatus infection [7]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.G138C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Multivariate analysis of overall survival or disease-free survival assay
• Mechanism Description: Each single mutationad a measurable effect on the affinity of the target enzyme for specific azole derivatives.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.G54E
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: Itraconazole resistance has been tightly linked to cyp51A mutations in the codon for Gly54, resulting in five different amino substitutions (G54k, G54V, G54R, G54E, and G54W).

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.G54K
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: The G54k amino acid change conferred cross-resistance to both itraconazole and posaconazole (26). The replacement of the wild-type chromosomal cyp51A allele by mutant allele bearing the G161A nucleotide change in codon 54 led to the acquisition of resistance to itraconazole de novo.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.P216L
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.G434C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.Y431C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [10]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Posaconazole
• Molecule Alteration: Missense mutation; p.Y121F+p.T289A+p.G448S+p.M172I
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain TR463; 746128
• Experiment for Molecule Alteration: PCR analysis
• Mechanism Description: In addition, to compare the susceptibility of TR463 with those of TR34 and TR46, the high resistance of TR463/Y121F/M172I/T289A/G448S was confirmed by MIC testing, displaying a pan-triazole-resistant phenotype to posaconazole, itraconazole, and voriconazole, indicating no in vitro activity of itraconazole and voriconazole (MIC, >16 mg/liter).

Voriconazole

Drug Resistance Data Categorized by Their Corresponding Mechanisms

Aberration of the Drug's Therapeutic Target (ADTT)

Disease Class: Aspergillus fumigatus infection [7]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.G138C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Multivariate analysis of overall survival or disease-free survival assay
• Mechanism Description: Each single mutationad a measurable effect on the affinity of the target enzyme for specific azole derivatives.

Disease Class: Aspergillus fumigatus infection [15]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.G448S
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: Resistance to voriconazole due to a G448S substitution in Aspergillus fumigatus in a patient with cerebral aspergillosis.

Disease Class: Aspergillus fumigatus infection [8]

• Resistant Disease: Aspergillus fumigatus infection [ICD-11: 1F20.2]
• Resistant Drug: Voriconazole
• Molecule Alteration: Tandem repeat; TR53 (GAATCACGCGGTCCGATGTGTGCTGAGCCGAATGAAAGTTGTCTAATGTCTAGAATCACGCGGTCCGATGTGTGCTGAGCCGAATGAAAGTTGTCTAATGTCTA)
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: RT-PCR
• Experiment for Drug Resistance: Broth microdilution method assay
• Mechanism Description: The azole-resistant A. fumigatus strains were detected tandem repeats (TRs) in the promoter region.

Disease Class: Invasive aspergillosis [1]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.G54R
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain RIT; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: NCCLS M38-P microdilution methodology assay
• Mechanism Description: Itraconazole resistance has been tightly linked to cyp51A mutations in the codon for Gly54, resulting in five different amino substitutions (G54k, G54V, G54R, G54E, and G54W).

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.H147Y
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.G434C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [9]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.Y431C
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain; 746128
• Experiment for Molecule Alteration: DNA sequencing assay
• Experiment for Drug Resistance: EUCAST method assay
• Mechanism Description: Four novel mutations were found (H147Y, P216L, Y431C, and G434C). The isolate bearing the P216L mutation was resistant to itraconazole and posaconazole, whereas the isolates with Y431C and G434C showed pan-azole resistance phenotypes.

Disease Class: Invasive aspergillosis [10]

• Resistant Disease: Invasive aspergillosis [ICD-11: 1F20.6]
• Resistant Drug: Voriconazole
• Molecule Alteration: Missense mutation; p.Y121F+p.T289A+p.G448S+p.M172I
• Experimental Note: Identified from the Human Clinical Data
• In Vitro Model: Aspergillus fumigatus strain TR463; 746128
• Experiment for Molecule Alteration: PCR analysis
• Mechanism Description: In addition, to compare the susceptibility of TR463 with those of TR34 and TR46, the high resistance of TR463/Y121F/M172I/T289A/G448S was confirmed by MIC testing, displaying a pan-triazole-resistant phenotype to posaconazole, itraconazole, and voriconazole, indicating no in vitro activity of itraconazole and voriconazole (MIC, >16 mg/liter).

References

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• Ref 10: A Novel Environmental Azole Resistance Mutation in Aspergillus fumigatus and a Possible Role of Sexual Reproduction in Its Emergence. mBio. 2017 Jun 27;8(3):e00791-17. doi: 10.1128/mBio.00791-17.
• Ref 11: Interrogation of related clinical pan-azole-resistant Aspergillus fumigatus strains: G138C, Y431C, and G434C single nucleotide polymorphisms in cyp51A, upregulation of cyp51A, and integration and activation of transposon Atf1 in the cyp51A promoter. Antimicrob Agents Chemother. 2011 Nov;55(11):5113-21. doi: 10.1128/AAC.00517-11. Epub 2011 Aug 29.
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• Ref 14: Mutations in Aspergillus fumigatus resulting in reduced susceptibility to posaconazole appear to be restricted to a single amino acid in the cytochrome P450 14alpha-demethylase. Antimicrob Agents Chemother. 2003 Feb;47(2):577-81. doi: 10.1128/AAC.47.2.577-581.2003.
• Ref 15: Resistance to voriconazole due to a G448S substitution in Aspergillus fumigatus in a patient with cerebral aspergillosis. J Clin Microbiol. 2012 Jul;50(7):2531-4. doi: 10.1128/JCM.00329-12. Epub 2012 May 9.