Deoxyhypusine synthase; Epilepsy; Hypusine; Neurodevelopmental disorder; Zebrafish; deoxyhypusine synthase; Oxidoreductases Acting on CH-NH Group Donors; Zebrafish Proteins; Animals; Zebrafish Proteins/metabolism; Zebrafish Proteins/genetics; Zebrafish Proteins/deficiency; Gene Knockdown Techniques; Phenotype; Brain/pathology; Brain/metabolism; Disease Models, Animal; Oxidoreductases Acting on CH-NH Group Donors/genetics; Oxidoreductases Acting on CH-NH Group Donors/metabolism; Epilepsy/genetics; Epilepsy/pathology; Epilepsy/physiopathology; Interneurons/metabolism; Brain; Interneurons; Molecular Biology; Cellular and Molecular Neuroscience
Abstract :
[en] DHPS deficiency syndrome is an ultra-rare neurodevelopmental disorder (NDD) which results from biallelic mutations in the gene encoding the enzyme deoxyhypusine synthase (DHPS). DHPS is essential to synthesize hypusine, a rare amino acid formed by post-translational modification of a conserved lysine in eukaryotic initiation factor 5 A (eIF5A). DHPS deficiency syndrome causes epilepsy, cognitive and motor impairments, and mild facial dysmorphology. In mice, a brain-specific genetic deletion of Dhps at birth impairs eIF5AHYP-dependent mRNA translation. This alters expression of proteins required for neuronal development and function, and phenotypically models features of human DHPS deficiency. We studied the role of DHPS in early brain development using a zebrafish loss-of-function model generated by knockdown of dhps expression with an antisense morpholino oligomer (MO) targeting the exon 2/intron 2 (E2I2) splice site of the dhps pre-mRNA. dhps knockdown embryos exhibited dose-dependent developmental delay and dysmorphology, including microcephaly, axis truncation, and body curvature. In dhps knockdown larvae, electrophysiological analysis showed increased epileptiform activity, and confocal microscopy analysis revealed reduced arborisation of GABAergic neurons. Our findings confirm that hypusination of eIF5A by DHPS is needed for early brain development, and zebrafish with an antisense knockdown of dhps model features of DHPS deficiency syndrome.
Disciplines :
Genetics & genetic processes
Author, co-author :
Shojaeinia, Elham; Center for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway ; Institute for Orphan Drug Discovery, Bremerhaven, Germany
Mastracci, Teresa L; Department of Biology, Indiana University-Indianapolis, Indianapolis, IN, USA
SOLIMAN, Remon ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB) > Molecular and Functional Neurobiology
Devinsky, Orrin; Department of Neurology, New York University Langone Medical Center, New York, NY, USA
Esguerra, Camila V; Center for Molecular Medicine Norway (NCMM), University of Oslo, Oslo, Norway
Crawford, Alexander D; Institute for Orphan Drug Discovery, Bremerhaven, Germany. crawford@biodiscoveryinstitute.org ; Luxembourg Centre for Systems Biomedicine (LCSB), University of Luxembourg, Belvaux, Luxembourg. crawford@biodiscoveryinstitute.org
External co-authors :
yes
Language :
English
Title :
Deoxyhypusine synthase deficiency syndrome zebrafish model: aberrant morphology, epileptiform activity, and reduced arborization of inhibitory interneurons.
M.H. Park E.C. Wolff Hypusine, a polyamine-derived amino acid critical for eukaryotic translation J Biol Chem 2018 10.1074/jbc.TM118.003341 30455349 6341376
Y. Zhang D. Su J. Zhu et al. Oxygen level regulates N-terminal translation elongation of selected proteins through deoxyhypusine hydroxylation Cell Rep 2022 10.1016/j.celrep.2022.110855 36640361 9875382
L.R. Padgett M.R. Shinkle S. Rosario et al. Deoxyhypusine synthase mutations alter the post-translational modification of eukaryotic initiation factor 5A resulting in impaired human and mouse neural homeostasis HGG Adv 2023 10.1016/j.xhgg.2023.100206 37333770 10275725
L.R. Padgett M.A. Robertson E.K. Anderson-Baucum et al. Deoxyhypusine synthase, an essential enzyme for hypusine biosynthesis, is required for proper exocrine pancreas development FASEB J 2021 10.1096/fj.201903177R 33811703
C.T. Connors C.B.P. Villaca E.K. Anderson-Baucum et al. A translational regulatory mechanism mediated by hypusinated eukaryotic initiation factor 5A facilitates β-cell identity and function Diabetes 2024 10.2337/db23-0148 38055903
A.P. Schuller C.C. Wu T.E. Dever A.R. Buskirk R. Green eIF5A functions globally in translation elongation and termination Mol Cell 2017 10.1016/j.molcel.2017.03.003 28575655 5414311
V. Pelechano P. Alepuz eIF5A facilitates translation termination globally and promotes the elongation of many non polyproline-specific tripeptide sequences Nucleic Acids Res 2017 10.1093/nar/gkx479 28637236 5737610
E. Gutierrez B.S. Shin C.J. Woolstenhulme et al. eIF5A promotes translation of polyproline motifs Mol Cell 2013 10.1016/j.molcel.2013.04.021 23727016 3744875
M. Hoque J.Y. Park Y.J. Chang et al. Regulation of gene expression by translation factor eIF5A: Hypusine-modified eIF5A enhances nonsense-mediated mRNA decay in human cells Translation (Austin) 2017 10.1080/21690731.2017.1366294 29034140
T. Abe R. Nagai S. Shimazaki et al. In vitro yeast reconstituted translation system reveals function of eIF5A for synthesis of long polypeptide J Biochem 2020 10.1093/jb/mvaa022 32251518
T. Li B. Belda-Palazón A. Ferrando P. Alepuz Fertility and polarized cell growth depends on eIF5A for translation of polyproline-rich formins in Saccharomyces cerevisiae Genetics 2014 10.1534/genetics.114.166926 25543667 4301661
P. Pálfi L. Bakacsy H. Kovács Á. Szepesi Hypusination, a metabolic posttranslational modification of eIF5A in plants during development and environmental stress responses Plants 2021 10.3390/plants1007126 34206171 8309165
Y. Liang C. Piao C.B. Beuschel et al. eIF5A hypusination, boosted by dietary spermidine, protects from premature brain aging and mitochondrial dysfunction Cell Rep 2021 10.1016/j.celrep.2021.108941 35026155 9000920
S.J. Hofer Y. Liang A. Zimmermann et al. Spermidine-induced hypusination preserves mitochondrial and cognitive function during aging Autophagy 2021 10.1080/15548627.2021.1933299 34105442 8386697
D.J. Puleston F. Baixauli D.E. Sanin et al. Polyamine metabolism is a central determinant of helper T cell lineage fidelity Cell 2021 10.1016/j.cell.2021.06.007 34216540 8358979
E. Anderson-Baucum A.R. Piñeros A. Kulkarni et al. Deoxyhypusine synthase promotes a pro-inflammatory macrophage phenotype Cell Metabol 2021 10.1016/j.cmet.2021.08.003
A.P. Sfakianos R.M. Raven A.E. Willis The pleiotropic roles of eIF5A in cellular life and its therapeutic potential in cancer Biochem Soc Trans 2022 10.1042/BST20221035 36511302 9788402
V. Faundes M.D. Jennings S. Crilly et al. Impaired eIF5A function causes a mendelian disorder that is partially rescued in model systems by spermidine Nat Commun 2021 10.1038/s41467-021-21053-2 33547280 7864902
A. Ziegler K. Steindl A.S. Hanner et al. Bi-allelic variants in DOHH, catalyzing the last step of hypusine biosynthesis, are associated with a neurodevelopmental disorder Am J Hum Genet 2022 10.1016/j.ajhg.2022.06.010 35858628 9388783
M. Ganapathi L.R. Padgett K. Yamada et al. Recessive rare variants in Deoxyhypusine synthase, an enzyme involved in the synthesis of Hypusine, are Associated with a neurodevelopmental disorder Am J Hum Genet 2019 10.1016/j.ajhg.2018.12.017 30661771 6369575
K. Nishimura S.B. Lee J.H. Park M.H. Park Essential role of eIF5A-1 and deoxyhypusine synthase in mouse embryonic development Amino Acids 2012 10.1007/s00726-011-0986-z 21850436
H. Sievert N. Pällmann K.K. Miller et al. A novel mouse model for inhibition of DOHH-mediated hypusine modification reveals a crucial function in embryonic development, proliferation and oncogenic transformation Dis Model Mech 2014 10.1242/dmm.014449 24832488 4107325
R.K. Kar A.S. Hanner M.F. Starost et al. Neuron-specific ablation of eIF5A or deoxyhypusine synthase leads to impairments in growth, viability, neurodevelopment, and cognitive functions in mice J Biol Chem 2021 10.1016/j.jbc.2021.101333 34688659 8605248
C.M. Carvalho S. Vasanth M. Shinawi et al. Dosage changes of a segment at 17p13.1 lead to intellectual disability and microcephaly as a result of complex genetic interaction of multiple genes Am J Hum Genet 2014 10.1016/j.ajhg.2014.10.006 25439725 4225592
T.L. Mastracci M.A. Robertson R.G. Mirmira R.M. Anderson Polyamine biosynthesis is critical for growth and differentiation of the pancreas Sci Rep 2015 10.1038/srep13269 26299433 4547391
A.P. Gobert J.L. Finley Y.L. Latour et al. Hypusination orchestrates the Antimicrobial response of macrophages Cell Rep 2020 10.1016/j.celrep.2020.108510 33326776 7812972
O. Crawley B. Grill Autophagy in axonal and presynaptic development Curr Opin Neurobiol 2021 10.1016/j.conb.2021.03.011 33940492 8387345
H. Igarashi J.G. Knott R.M. Schultz C.J. Williams Alterations of PLCbeta1 in mouse eggs change calcium oscillatory behavior following fertilization Dev Biol 2007 10.1016/j.ydbio.2007.09.028 17961538 2170533
T. Afrikanova A.S. Serruys O.E. Buenafe et al. Validation of the zebrafish pentylenetetrazol seizure model: locomotor versus electrographic responses to antiepileptic drugs PLoS ONE 2013 10.1371/journal.pone.0054166 24349101 3862488
M. Zijlmans P. Jiruska R. Zelmann F.S. Leijten J.G. Jefferys J. Gotman High-frequency oscillations as a new biomarker in epilepsy Ann Neurol 2012 10.1002/ana.22548 22367988 3754947
E. Tiraboschi S. Martina W. van der Ent et al. New insights into the early mechanisms of epileptogenesis in a zebrafish model of Dravet syndrome Epilepsia 2020 10.1111/epi.16456 32096222
T.A. Ferreira A.V. Blackman J. Oyrer et al. Neuronal morphometry directly from bitmap images Nat Methods 2014 10.1038/nmeth.3125 25264773 5271921
D. Ristanović N.T. Milosević V. Stulić Application of modified Sholl analysis to neuronal dendritic arborization of the cat spinal cord J Neurosci Methods 2006 10.1016/j.jneumeth.2006.05.030 16814868
