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Drosophila carrying epilepsy-associated variants in the vitamin B6 metabolism gene PNPO display allele- and diet-dependent phenotypes
Journal article   Open access   Peer reviewed

Drosophila carrying epilepsy-associated variants in the vitamin B6 metabolism gene PNPO display allele- and diet-dependent phenotypes

Wanhao Chi, Atulya S R Iyengar, Wenqin Fu, Wei Liu, Abigayle E Berg, Chun-Fang Wu and Xiaoxi Zhuang
Proceedings of the National Academy of Sciences - PNAS, Vol.119(9), e2115524119
03/01/2022
DOI: 10.1073/pnas.2115524119
PMCID: PMC8892510
PMID: 35217610
url
https://doi.org/10.1073/pnas.2115524119View
Published (Version of record) Open Access

Abstract

Pyridox(am)ine 5 -phosphate oxidase (PNPO) catalyzes the rate-limiting step in the synthesis of pyridoxal 5 -phosphate (PLP), the active form of vitamin B6 required for the synthesis of neurotransmitters gamma-aminobutyric acid (GABA) and the monoamines. Pathogenic variants in have been increasingly identified in patients with neonatal epileptic encephalopathy and early-onset epilepsy. These patients often exhibit different types of seizures and variable comorbidities. Recently, the gene has also been implicated in epilepsy in adults. It is unclear how these phenotypic variations are linked to specific alleles and to what degree diet can modify their expression. Using CRISPR-Cas9, we generated four knock-in alleles, , , , and , in which the endogenous was replaced by wild-type human complementary DNA (cDNA) and three epilepsy-associated variants. We found that these knock-in flies exhibited a wide range of phenotypes, including developmental impairments, abnormal locomotor activities, spontaneous seizures, and shortened life span. These phenotypes are allele dependent, varying with the known biochemical severity of these mutations and our characterized molecular defects. We also showed that diet treatments further diversified the phenotypes among alleles, and PLP supplementation at larval and adult stages prevented developmental impairments and seizures in adult flies, respectively. Furthermore, we found that h had a significant dominant-negative effect, rendering heterozygous flies susceptible to seizures and premature death. Together, these results provide biological bases for the various phenotypes resulting from multifunction of , specific molecular and/or genetic properties of each variant, and differential allele-diet interactions.
vitamin B6 sugarlethal gene–environment interaction pyridox(am)ine 5′-phosphate oxidase phenotypic variation

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