Abstract
We aimed to detect the causative gene in five unrelated families with recessive inheritance pattern neurological disorders involving the central nervous system, and the potential function of the NEMF gene in the central nervous system. Exome sequencing (ES) was applied to all families and linkage analysis was performed on family 1. A minigene assay was used to validate the splicing effect of the relevant discovered variants. Immunofluorescence (IF) experiment was performed to investigate the role of the causative gene in neuron development. The large consanguineous family confirms the phenotype-causative relationship with homozygous frameshift variant (NM_004713.6:c.2618del) as revealed by ES. Linkage analysis of the family showed a significant single-point LOD of 4.5 locus. Through collaboration in GeneMatcher, four additional unrelated families’ likely pathogenic NEMF variants for a spectrum of central neurological disorders, two homozygous splice-site variants (NM_004713.6:c.574+1G>T and NM_004713.6:c.807-2A>C) and a homozygous frameshift variant (NM_004713.6: c.1234_1235insC) were subsequently identified and segregated with all affected individuals. We further revealed that knockdown (KD) of Nemf leads to impairment of axonal outgrowth and synapse development in cultured mouse primary cortical neurons. Our study demonstrates that disease-causing biallelic NEMF variants result in central nervous system impairment and other variable features. NEMF is an important player in mammalian neuron development.
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Abbreviations
- AR:
-
Autosomal recessive
- RoH:
-
Regions of homozygosity
- LOD:
-
Log of the odds
- shRNA:
-
Short hairpin RNA
- ID:
-
Intellectual disability
- KD:
-
Knockdown
- ES:
-
Exome sequencing
- IF:
-
Immunofluorescence
- PBS:
-
Phosphate-buffered saline
- DD:
-
Developmental delay
- RQC:
-
Ribosome-associated quality control
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Acknowledgements
We are especially grateful to the families that participated in this study. We thank Tomasz J. Nowakowski for sharing the single-cell RNA-seq data of the human developing cerebral cortex.
Funding
The following grants supported this work: the National Natural Science Foundation of China (81671122) and the Science and Technology Major Projects of Hunan Province Science and Technology Department (2018DK2016) to Zhengmao Hu.; the National Natural Science Foundation of China (81730036, 81525007) and the Science and Technology Major Projects of Hunan Province Science and Technology Department (2018SK1030) to Kun Xia.; the National Natural Science Foundation of China (31671114, 81871079) to Hui Guo; Hainan Provincial Department of Science and Technology (CN) (2016RS2001, 2016JC2055).
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Ahmed, A., Wang, M., Bergant, G. et al. Biallelic loss-of-function variants in NEMF cause central nervous system impairment and axonal polyneuropathy. Hum Genet 140, 579–592 (2021). https://doi.org/10.1007/s00439-020-02226-3
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DOI: https://doi.org/10.1007/s00439-020-02226-3