Eif2b3 mutants recapitulate phenotypes of Vanishing White Matter Disease and validate novel disease alleles in zebrafish

Lee, Y.R., Kim, S.H., Ben-Mahmoud, A., Kim, O.H., Choi, T.I., Lee, K.H., Ku, B., Eum, J., Kee, Y., Lee, S., Cha, J., Won, D., Lee, S.T., Choi, J.R., Lee, J.S., Kim, H.D., Kim, H.G., Bonkowsky, J.L., Kang, H.C., Kim, C.H.
Human molecular genetics   30(5): 331-342 (Journal)
Registered Authors
Bonkowsky, Joshua, Kee, Yun, Kim, Cheol-Hee
MeSH Terms
  • Alleles
  • Animals
  • Cell Differentiation
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Disease Models, Animal
  • Eukaryotic Initiation Factor-2B/chemistry
  • Eukaryotic Initiation Factor-2B/genetics*
  • Female
  • Gene Expression Regulation, Developmental*
  • Gene Knockout Techniques
  • Humans
  • Infant
  • Leukoencephalopathies/genetics*
  • Leukoencephalopathies/metabolism
  • Models, Molecular
  • Myelin Sheath/genetics*
  • Myelin Sheath/metabolism
  • Neovascularization, Physiologic*/genetics
  • Protein Conformation
  • Sequence Deletion
  • Stress, Physiological
  • Vascular Endothelial Growth Factor A/metabolism
  • Zebrafish/genetics*
  • Zebrafish/metabolism*
33517449 Full text @ Hum. Mol. Genet.
Leukodystrophy with Vanishing White Matter (VWM), also called Childhood Ataxia with Central Nervous System Hypomyelination (CACH), is caused by mutations in the subunits of the eukaryotic translation initiation factor, EIF2B1, EIF2B2, EIF2B3, EIF2B4, or EIF2B5. However, little is known regarding the underlying pathogenetic mechanisms, and there is no curative treatment for VWM. In this study, we established the first EIF2B3 animal model for VWM disease in vertebrates by CRISPR mutagenesis of the highly conserved zebrafish ortholog eif2b3. Using CRISPR, we generated two mutant alleles in zebrafish eif2b3, 10- and 16-bp deletions, respectively. The eif2b3 mutants showed defects in myelin development and glial cell differentiation, and increased expression of genes in the induced stress response pathway. Interestingly, we also found ectopic angiogenesis and increased VEGF expression. Ectopic angiogenesis in the eif2b3 mutants was reduced by administration of VEGF receptor inhibitor SU5416. Using the eif2b3 mutant zebrafish model together with in silico protein modeling analysis, we demonstrated the pathogenicity of 18 reported mutations in EIF2B3, as well as of a novel variant identified in a 19-month-old female patient: c.503 T > C (p.Leu168Pro). In summary, our zebrafish mutant model of eif2b3 provides novel insights into VWM pathogenesis and offers rapid functional analysis of human EIF2B3 gene variants.
Genes / Markers
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Engineered Foreign Genes
Errata and Notes