Biochemical Characterization of AGMO Variants Implicated in Relapses in Visceral Leishmaniasis

Watschinger, Katrin; Keller, Markus A.; Golderer, Georg; Coassin, Stefan; Zschocke, Johannes; Werner, Ernst R.

doi:10.1093/infdis/jiy090

Cited by 4 publications

(2 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…From five Sudanese families (10 individuals, two of each family) with visceral leishmania (Kala-azar) relapses, rare mutations in the AGMO gene were found, of which variant 1 (c.701A>G, rs143439626, p.Lys234Arg) led to an amino acid exchange in exon 7 and variant 2 led to a stop in exon 12 (c.1213C>T, rs139309795, p.Arg405Ter) [ 83 ]. Both mutations were biochemically characterised, and results confirmed a loss of function only for mutation 2 (p.Arg405Ter), whereas the AGMO protein with mutation 1 (p.Lys234Arg) was proven to be functionally active in a recombinant expression model [ 84 ]. So far, it is not fully clear whether disrupted AGMO function alone or together with other mutated genes is causative for the recurrence of visceral leishmania and therefore needs to be further assessed.…”

Section: Agmo In Human Diseasesmentioning

confidence: 99%

The Emerging Physiological Role of AGMO 10 Years after Its Gene Identification

Sailer

Keller

Werner

et al. 2021

Life

Self Cite

View full text Add to dashboard Cite

The gene encoding alkylglycerol monooxygenase (AGMO) was assigned 10 years ago. So far, AGMO is the only known enzyme capable of catalysing the breakdown of alkylglycerols and lyso-alkylglycerophospholipids. With the knowledge of the genetic information, it was possible to relate a potential contribution for mutations in the AGMO locus to human diseases by genome-wide association studies. A possible role for AGMO was implicated by genetic analyses in a variety of human pathologies such as type 2 diabetes, neurodevelopmental disorders, cancer, and immune defence. Deficient catabolism of stored lipids carrying an alkyl bond by an absence of AGMO was shown to impact on the overall lipid composition also outside the ether lipid pool. This review focuses on the current evidence of AGMO in human diseases and summarises experimental evidence for its role in immunity, energy homeostasis, and development in humans and several model organisms. With the progress in lipidomics platform and genetic identification of enzymes involved in ether lipid metabolism such as AGMO, it is now possible to study the consequence of gene ablation on the global lipid pool and further on certain signalling cascades in a variety of model organisms in more detail.

show abstract

Section: Agmo In Human Diseasesmentioning

confidence: 99%

The Emerging Physiological Role of AGMO 10 Years after Its Gene Identification

Sailer

Keller

Werner

et al. 2021

Life

Self Cite

View full text Add to dashboard Cite

show abstract

“…Additionally, AGMO might play a role in inflammation [ 14 ]. Mutations in the AGMO gene were found to be associated with visceral leishmania relapse in humans [ 15 ], but other genes might also be involved [ 16 ]. A role of AGMO in platelet-activating factor regulation was suggested [ 17 ], but could not be reproduced in our cell system [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

When the genome bluffs: a tandem duplication event during generation of a novel Agmo knockout mouse model fools routine genotyping

et al. 2021

Self Cite

View full text Add to dashboard Cite

Background Genome editing in mice using either classical approaches like homologous recombination or CRISPR/Cas9 has been reported to harbor off target effects (insertion/deletion, frame shifts or gene segment duplications) that lead to mutations not only in close proximity to the target site but also outside. Only the genomes of few engineered mouse strains have been sequenced. Since the role of the ether-lipid cleaving enzyme alkylglycerol monooxygenase (AGMO) in physiology and pathophysiology remains enigmatic, we created a knockout mouse model for AGMO using EUCOMM stem cells but unforeseen genotyping issues that did not agree with Mendelian distribution and enzyme activity data prompted an in-depth genomic validation of the mouse model. Results We report a gene segment tandem duplication event that occurred during the generation of an Agmo knockout-first allele by homologous recombination. Only low homology was seen between the breakpoints. While a single copy of the recombinant 18 kb cassette was integrated correctly around exon 2 of the Agmo gene, whole genome nanopore sequencing revealed a 94 kb duplication in the Agmo locus that contains Agmo wild-type exons 1–3. The duplication fooled genotyping by routine PCR, but could be resolved using qPCR-based genotyping, targeted locus amplification sequencing and nanopore sequencing. Despite this event, this Agmo knockout mouse model lacks AGMO enzyme activity and can therefore be used to study its physiological role. Conclusions A duplication event occurred at the exact locus of the homologous recombination and was not detected by conventional quality control filters such as FISH or long-range PCR over the recombination sites. Nanopore sequencing provides a cost convenient method to detect such underrated off-target effects, suggesting its use for additional quality assessment of gene editing in mice and also other model organisms.

show abstract