Clinical and experimental phenotype of azole-resistant Aspergillus fumigatus with a HapE splice site mutation: a case report

Abstract Background The recent increase in cases of azole-resistant Aspergillus fumigatus (ARAf) infections is a major clinical concern owing to its treatment limitations. Patient-derived ARAf occurs after prolonged azole treatment in patients with aspergillosis and involves various cyp51A point mutations or non-cyp51A mutations. The prognosis of patients with chronic pulmonary aspergillosis (CPA) with patient-derived ARAf infection remains unclear. In this study, we reported the case of a patient with ARAf due to HapE mutation, as well as the virulence of the isolate. Case presentation A 37-year-old male was presented with productive cough and low-grade fever. The patient was diagnosed with CPA based on the chronic course, presence of a fungus ball in the upper left lobe on chest computed tomography (CT), positivity for Aspergillus-precipitating antibody and denial of other diseases. The patient underwent left upper lobe and left S6 segment resection surgery because of repeated haemoptysis during voriconazole (VRC) treatment. The patient was postoperatively treated with VRC for 6 months. Since then, the patient was followed up without antifungal treatment but relapsed 4 years later, and VRC treatment was reinitiated. Although an azole-resistant isolate was isolated after VRC treatment, the patient did not show any disease progression in either respiratory symptoms or radiological findings. The ARAf isolated from this patient showed slow growth, decreased biomass and biofilm formation in vitro, and decreased virulence in the Galleria mellonella infection model compared with its parental strain. These phenotypes could be caused by the HapE splice site mutation. Conclusions This is the first to report a case demonstrating the clinical manifestation of a CPA patient infected with ARAf with a HapE splice site mutation, which was consistent with the in vitro and in vivo attenuated virulence of the ARAf isolate. These results imply that not all the ARAf infections in immunocompetent patients require antifungal treatment. Further studies on the virulence of non-cyp51A mutations in ARAf are warranted.

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A splice site mutation of alpha-spectrin gene causing skipping of exon 18 in hereditary elliptocytosis

Blood ◽

10.1182/blood.v81.10.2791.2791 ◽

1993 ◽

Vol 81 (10) ◽

pp. 2791-2798 ◽

Cited By ~ 16

Author(s):

N Alloisio ◽

R Wilmotte ◽

J Marechal ◽

P Texier ◽

L Denoroy ◽

...

Keyword(s):

Splice Site ◽

Splice Site Mutation ◽

Low Grade ◽

Site Mutation ◽

Acceptor Splice Site ◽

Genetic Lesion ◽

Major Disadvantage ◽

Internal Position ◽

Alpha Spectrin ◽

Self Association

Abstract Spectrin Oran (alpha II/21) has been reported previously as a variant of the alpha II domain. Its expression level is low (10% of total spectrin) in heterozygotes denoting a major disadvantage of the mutated alpha-chain dimer or tetramer with respect to their normal counterparts. Spectrin Oran is associated with symptomatic elliptocytosis in the homozygous state. A 1-minute digestion time allowed to perceive a fast trypsin cleavage (not existing normally) after Arg 890 (helix 3 of repeating segment alpha 9). The responsible change was the lack of amino acids 822 to 862 (helix 2 of repeating segment alpha 8). Such a situation fits with the phasing of spectrin according to which mutated helix 2 and distorted helix 3 are adjacent to one another. The internal position of the structural change accounts for the slight self-association defect. The ultimate genetic lesion was a G to A substitution (intronic position-1) in the acceptor splice site of intron 17 resulting in skipping of exon 18. The substitution also created an acceptor splice site 1 base downstream, but the latter was used at a low grade.

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Homozygous Splice Site Mutation in ZP1 Causes Familial Oocyte Maturation Defect

Genes ◽

10.3390/genes11040382 ◽

2020 ◽

Vol 11 (4) ◽

pp. 382 ◽

Cited By ~ 2

Author(s):

Özlem Okutman ◽

Cem Demirel ◽

Firat Tülek ◽

Veronique Pfister ◽

Umut Büyük ◽

...

Keyword(s):

Oocyte Maturation ◽

Splice Site ◽

Splice Site Mutation ◽

Controlled Ovarian Hyperstimulation ◽

Sequencing Analysis ◽

Site Mutation ◽

Vitro Fertilization ◽

Maturation Defect ◽

Exon 11

In vitro fertilization (IVF) involves controlled ovarian hyperstimulation using hormones to produce large numbers of oocytes. The success of IVF is tightly linked to the availability of mature oocytes. In most cases, about 70% to 80% of the oocytes are mature at the time of retrieval, however, in rare instances, all of them may be immature, implying that they were not able to reach the metaphase II (MII) stage. The failure to obtain any mature oocytes, despite a well conducted ovarian stimulation in repeated cycles is a very rare cause of primary female infertility, for which the underlying suspected genetic factors are still largely unknown. In this study, we present the whole exome sequencing analysis of a consanguineous Turkish family comprising three sisters with a recurrent oocyte maturation defect. Analysis of the data reveals a homozygous splice site mutation (c.1775-3C>A) in the zona pellucida glycoprotein 1 (ZP1) gene. Minigene experiments show that the mutation causes the retention of the intron 11 sequence between exon 11 and exon 12, resulting in a frameshift and the likely production of a truncated protein.

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Homozygous mutations in REC114 cause female infertility characterised by multiple pronuclei formation and early embryonic arrest

Journal of Medical Genetics ◽

10.1136/jmedgenet-2019-106379 ◽

2019 ◽

Vol 57 (3) ◽

pp. 187-194 ◽

Cited By ~ 2

Author(s):

Wenjing Wang ◽

Jie Dong ◽

Biaobang Chen ◽

Jing Du ◽

Yanping Kuang ◽

...

Keyword(s):

Missense Mutation ◽

Splice Site ◽

Genetic Factors ◽

Splice Site Mutation ◽

Female Infertility ◽

Site Mutation ◽

Whole Exome ◽

Pronuclear Formation ◽

Embryonic Arrest

BackgroundAbnormal pronuclear formation during fertilisation and subsequent early embryonic arrest results in female infertility. In recent years, with the prevalence of assisted reproductive technology, a few genes have been identified that are involved in female infertility caused by abnormalities in oocyte development, fertilisation and embryonic development. However, the genetic factors responsible for multiple pronuclei formation during fertilisation and early embryonic arrest remain largely unknown.ObjectiveWe aim to identify genetic factors responsible for multiple pronuclei formation during fertilisation or early embryonic arrest.MethodsWhole-exome sequencing was performed in a cohort of 580 patients with abnormal fertilisation and early embryonic arrest. Effects of mutations were investigated in HEK293T cells by western blotting and immunoprecipitation, as well as minigene assay.ResultsWe identified a novel homozygous missense mutation (c.397T>G, p.C133G) and a novel homozygous donor splice-site mutation (c.546+5G>A) in the meiotic gene REC114. REC114 is involved in the formation of double strand breaks (DSBs), which initiate homologous chromosome recombination. We demonstrated that the splice-site mutation affected the normal alternative splicing of REC114, while the missense mutation reduced the protein level of REC114 in vitro and resulted in the loss of its function to protect its partner protein MEI4 from degradation.ConclusionsOur study has identified mutations in REC114 responsible for human multiple pronuclei formation and early embryonic arrest, and these findings expand our knowledge of genetic factors that are responsible for normal human female meiosis and fertility.

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Correction to: Clinical and experimental phenotype of azole-resistant Aspergillus fumigatus with a HapE splice site mutation: a case report

BMC Infectious Diseases ◽

10.1186/s12879-021-06667-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yuya Ito ◽

Takahiro Takazono ◽

Satoru Koga ◽

Yuichiro Nakano ◽

Nobuyuki Ashizawa ◽

...

Keyword(s):

Case Report ◽

Aspergillus Fumigatus ◽

Splice Site ◽

Splice Site Mutation ◽

Site Mutation

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A splice site mutation of alpha-spectrin gene causing skipping of exon 18 in hereditary elliptocytosis

Blood ◽

10.1182/blood.v81.10.2791.bloodjournal81102791 ◽

1993 ◽

Vol 81 (10) ◽

pp. 2791-2798 ◽

Cited By ~ 1

Author(s):

N Alloisio ◽

R Wilmotte ◽

J Marechal ◽

P Texier ◽

L Denoroy ◽

...

Keyword(s):

Splice Site ◽

Splice Site Mutation ◽

Low Grade ◽

Site Mutation ◽

Acceptor Splice Site ◽

Genetic Lesion ◽

Major Disadvantage ◽

Internal Position ◽

Alpha Spectrin ◽

Self Association

Spectrin Oran (alpha II/21) has been reported previously as a variant of the alpha II domain. Its expression level is low (10% of total spectrin) in heterozygotes denoting a major disadvantage of the mutated alpha-chain dimer or tetramer with respect to their normal counterparts. Spectrin Oran is associated with symptomatic elliptocytosis in the homozygous state. A 1-minute digestion time allowed to perceive a fast trypsin cleavage (not existing normally) after Arg 890 (helix 3 of repeating segment alpha 9). The responsible change was the lack of amino acids 822 to 862 (helix 2 of repeating segment alpha 8). Such a situation fits with the phasing of spectrin according to which mutated helix 2 and distorted helix 3 are adjacent to one another. The internal position of the structural change accounts for the slight self-association defect. The ultimate genetic lesion was a G to A substitution (intronic position-1) in the acceptor splice site of intron 17 resulting in skipping of exon 18. The substitution also created an acceptor splice site 1 base downstream, but the latter was used at a low grade.

Download Full-text