Alternating Hemiplegia of Childhood (AHC) is a devastating autosomal dominant disorder caused by ATP1A3 mutations, resulting in severe hemiplegia and dystonia spells, ataxia, debilitating disabilities, and premature death. Here, we determine the effects of delivering an extra copy of the normal gene in a mouse model carrying the most common mutation causing AHC in humans, the D801N mutation. We used an adeno-associated virus serotype 9 (AAV9) vector expressing the human ATP1A3 gene under the control of a human Synapsin promoter. We first demonstrated that intracerebroventricular (ICV) injection of this vector in wild-type mice on postnatal day 10 (P10) results in increases in ouabain-sensitive ATPase activity and in expression of reporter genes in targeted brain regions. We then tested this vector in mutant mice. Simultaneous intracisterna magna and bilateral ICV injections of this vector at P10 resulted, at P40, in reduction of inducible hemiplegia spells, improvement in balance beam test performance, and prolonged survival of treated mutant mice up to P70. Our study demonstrates, as a proof of concept, that gene therapy can induce favorable effects in a disease caused by a mutation of the gene of a protein that is, at the same time, an ATPase enzyme, a pump, and a signal transduction factor.
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References
1.
FernandesC, MikatiMA. The expanding spectrum of ATP1A3 related disease. Eur J Paediatr Neurol, 2019; 23:345–346.
2.
MasoudM, PrangeL, WuchichJ, et al.Diagnosis and treatment of alternating hemiplegia of childhood. Curr Treat Options Neurol, 2017; 19:8.
3.
JasienJM, BonnerM, D'alliR, et al.Cognitive, adaptive, and behavioral profiles and management of alternating hemiplegia of childhood. Dev Med Child Neurol, 2019; 61:547–554.
4.
KansagraS, GhusayniR, KherallahB, et al.Polysomnography findings and sleep disorders in children with alternating hemiplegia of childhood. J Clin Sleep Med, 2019; 15:65–70.
5.
MasoudM, GordonK, HallA, et al.Motor function domains in alternating hemiplegia of childhood. Dev Med Child Neurol, 2017; 59:822–828.
6.
PanagiotakakiE, GobbiG, NevilleB, et al.Evidence of a non-progressive course of alternating hemiplegia of childhood: study of a large cohort of children and adults. Brain, 2010; 133:3598–3610.
7.
UchitelJ, AbdelnourE, BoggsA, et al.Social responsiveness in alternating hemiplegia if childhood. Dev Med Child Neurol 2020. 2020; 62:820–826.
8.
UchitelJ, HelsethA, PrangeL, et al.The epileptology of alternating hemiplegia of childhood. Neurology, 2019; 93:e1248–e1259.
9.
HeinzenEL, ArzimanoglouA, BrashearA, et al.ATP1A3 Working Group. Distinct neurological disorders with ATP1A3 mutations. Lancet Neurol, 2014; 13:503–514.
10.
HeinzenEL, SwobodaKJ, HitomiY, et al.De novo mutations in ATP1A3 cause alternating hemiplegia of childhood. Nat Genet, 2012; 44:1030–1034.
11.
PanagiotakakiE, De GrandisE, StagnaroM, et al.Clinical profile of patients with ATP1A3 mutations in Alternating Hemiplegia of Childhood-a study of 155 patients. Orphanet J Rare Dis, 2015; 10:123.
MikatiMA, KramerU, ZupancML, et al.Alternating hemiplegia of childhood: clinical manifestations and long-term outcome. Pediatr Neurol, 2000; 23:134–141.
14.
MikatiMA, MaguireH, BarlowCF, et al.A syndrome of autosomal dominant alternating hemiplegia: clinical presentation mimicking intractable epilepsy; chromosomal studies; and physiologic investigations. Neurology, 1992; 42:2251–2257.
15.
HelsethAR, HunanyanAS, AdilS, et al.Novel E815K knock-in mouse model of alternating hemiplegia of childhood. Neurobiol Dis, 2018; 119:100–112.
16.
HunanyanAS, FainbergNA, LinabargerM, et al.Knock-in mouse model of alternating hemiplegia of childhood: behavioral and electrophysiologic characterization. Epilepsia, 2015; 56:82–93.
17.
HunanyanAS, HelsethAR, AbdelnourE, et al.Mechanisms of increased hippocampal excitability in the Mashl+/− mouse model of Na+/K+-ATPase dysfunction. Epilepsia, 2018; 59:1455–1468.
18.
ClapcoteSJ, DuffyS, XieG, et al.Mutation I810N in the alpha3 isoform of Na+,K+-ATPase causes impairments in the sodium pump and hyperexcitability in the CNS. Proc Natl Acad Sci U S A, 2009:106:14085–14090.
19.
KirshenbaumGS, ClapcoteSJ, DuffyS, et al.Mania-like behavior induced by genetic dysfunction of the neuron-specific Na+,K+-ATPase α3 sodium pump. Proc Natl Acad Sci U S A, 2011; 108:18144–18149.
20.
KirshenbaumGS, DachtlerJ, RoderJC, et al.Transgenic rescue of phenotypic deficits in a mouse model of alternating hemiplegia of childhood. Neurogenetics, 2016; 17:57–63.
21.
MarshallMS, IssaY, JakubauskasB, et al.Long-term improvement of neurological signs and metabolic dysfunction in a mouse model of Krabbe's disease after global gene therapy. Mol Ther, 2018; 26:874–889.
22.
MendellJR, SahenkZ, MalikV, et al.A phase ½a follistatin gene therapy trial for Becker muscular dystrophy. Mol Ther, 2015; 23:192–201.
23.
NakamuraS, MuramatsuSI, TakinoN, et al.Gene therapy for Glut1-deficient mouse using an adeno-associated virus vector with the human intrinsic GLUT1 promoter. J Gene Med, 2018; 20:e3013.
24.
UchitelJ, KantorB, SmithEC, et al.Viral-mediated gene replacement therapy in the developing central nervous system: current status and future directions. Pediatr Neurol. 2020; 110:5–19.
25.
KomuraH, KakioS, SasaharaT, et al.Alzheimer Aβ assemblies accumulate in excitatory neurons upon proteasome inhibition and kill nearby NAKα3 neurons by secretion. iScience, 2019; 13:452–477.
26.
OhnishiT, YanazawaM, SasaharaT, et al.Na, K-ATPase α3 is a death target of Alzheimer patient amyloid-β assembly. Proc Natl Acad Sci U S A, 2015; 112:E4465–E4474.
27.
ShrivastavaAN, RedekerV, FritzN, et al.α-Synuclein assemblies sequester neuronal α3-Na+/K+-ATPase and impair Na+ gradient. EMBO J, 2015; 34:2408–2423.
28.
YinQ, LiuJJ, TianMT, et al.Protein fibrillation in neurodegenerative diseases and its chiral interaction with interfaces. Acta Polymer Sin, 2019; 50:575–587.
29.
Bravo-HernandezM, TadokoroT, NavarroMR, et al.Spinal subpial delivery of AAV9 enables widespread gene silencing and blocks motoneuron degeneration in ALS. Nat Med, 2020; 26:118–130.
30.
HammondSL, LeekAN, RichmanEH, et al.Cellular selectivity of AAV serotypes for gene delivery in neurons and astrocytes by neonatal intracerebroventricular injection. PloS One, 2017; 12:e0188830.
31.
ZhangH, YangB, MuX, et al.Several rAAV vectors efficiently cross the blood–brain barrier and transduce neurons and astrocytes in the neonatal mouse central nervous system. Mol Ther, 2011; 19:1440–1448.
32.
ZincarelliC, SoltysS, RengoG, et al.Analysis of AAV serotypes 1–9 medicated gene expression and tropism in mice after systemic injection. Mol Ther, 2008; 16:1073–1080.
33.
PattaliR, MouY, LiXJ. AAV9 vector: a novel modality in gene therapy for spinal muscular atrophy. Gene Ther, 2019; 26:287–295.
34.
BøttgerP, TraczZ, HeuckA, et al.Distribution of Na/K-ATPase alpha 3 isoform, a sodium-potassium P-type pump associated with rapid-onset of dystonia parkinsonism (RDP) in the adult mouse brain. J Comp Neurol, 2011; 519:376–404.
35.
MurataK, KinoshitaT, IshikawaT, et al.Region- and neuronal-subtype-specific expression of Na,K-ATPase alpha and beta subunit isoforms in the mouse brain. J Comp Neurol, 2020; 528:2654–2678.
36.
LarsenBR, StoicaA, MacAulayN. Managing brain extracellular K(+) during neuronal activity: the physiological role of the Na(+)/K(+)-ATPase subunit isoforms. Front Physiol, 2016; 7:141.
37.
MonahanPE, LothropCD, SunJ, et al.Proteasome inhibitors enhance gene delivery by AAV virus vectors expressing large genomes in hemophilia mouse and dog models: a strategy for broad clinical application. Mol Ther, 2010; 18:1907–1196.
38.
RobbinsKL, GlascockJJ, OsmanEY, et al.Defining the therapeutic window in a severe animal model of spinal muscular atrophy. Hum Mol Genet, 2014; 23:4559–4568.
39.
OliveiraMS, FurianAF, RamboLM, et al.Prostaglandin E2 modulates Na,K-ATPase activity in rat hippocampus: implications for neurological diseases. J Neurochem, 2009; 109:416–426.
40.
ZeiniehMP, TalhoukRS, El-SabbanME, et al.Differential expression of hippocampal connexins after acute hypoxia in the developing brain. Brain Dev, 2010; 32:810–817.
41.
RoosingS, HofreeM, KimS, et al.Functional genome-wide siRNA screen identifies KIAA0586 as mutated in Joubert syndrome. Elife, 2015; 4:e06602.
42.
IsaksenTJ, KrosL, VedovatoN, et al.Hypothermia-induced dystonia and abnormal cerebellar activityin a mouse model with a single disease-mutation in the sodium-potassium pump. PloS Genet, 2017; 13:e1006763.
43.
MikatiMA, HolmesGL, ChronopoulosA, et al.Phenobarbital modifies seizure-related brain injury in the developing brain. Ann Neurol, 1994; 36:425–433.
44.
PickCG, YanaiJ. Long-term reduction in spontaneous alternations after early exposure to phenobarbital. Int J Dev Neurosci, 1984; 2:223–228.
45.
KeelerAM, ZiegerM, TodeasaSH, et al.Systemic delivery of AAVB1-GAA clears glycogen and prolongs survival in a mouse model of Pompe disease. Hum Gene Ther, 2019; 30:57–68.
46.
FunckVR, RibeiroLR, PereiraLM, et al.Long-term decrease in Na+,K+-ATPase activity after pilocarpine-induced status epilepticus is associated with nitration of its alpha subunit. Epilepsy Res, 2014; 108:1705–1710.
47.
ThorntonC, LeawB, MallardC, et al.Cell death in the developing brain after hypoxia-ischemia. Front Cell Neurosci, 2017; 11:248.
48.
ShrivastavaAN, TrillerA, MelkiR. Cell biology and dynamics of Neuronal Na+/K+-ATPase in health and diseases. Neuropharmacology, 2020; 169:107461.
49.
AhlfeldJ, FilserS, SchmidtF, et al.Neurogenesis from Sox2 expressing cells in the adult cerebellar cortex. Sci Rep, 2017; 7:6137.
ColellaP, SellierP, Costa VerderaH, et al.AAV gene transfer with tandem promoter design prevents anti-transgene immunity and provides persistent efficacy in neonate pompe mice. Mol Ther Methods Clin Dev, 2018; 12:85–101.
52.
De CamilliP, CameronR, GreengardP. Synapsin I (protein I), a nerve terminal-specific phosphoprotein. I. Its general distribution in synapses of the central and peripheral nervous system demonstrated by immunofluorescence in frozen and plastic sections. J Cell Biol, 1983; 96:1337–1354.
53.
KantorB, TagliafierroL, GuJ, et al.Downregulation of SNCA expression by targeted editing of DNA methylation: a potential strategy for precision therapy in PD. Mol Ther, 2018; 26:2638–2649.
54.
LykkenEA, ShyngC, EdwardsRJ, et al.Recent progress and considerations for AAV gene therapies targeting the central nervous system. J Neurodev Disord, 2018; 10:16.
55.
ReinhardL, TidowH, ClausenMJ, et al.Na(+),K (+)-ATPase as a docking station: protein-protein complexes of the Na(+),K (+)-ATPase. Cell Mol Life Sci, 2013; 70:205–222.
56.
LiM, JazayeriD, CorryB, et al.A functional correlate of severity in alternating hemiplegia of childhood. Neurobiol Dis, 2015; 77:88–93.
57.
HoltRL, ArehartE, HunanyanA, et al.Pediatric sudden unexpected death in epilepsy: what have we learned from animal and human studies, and can we prevent it?. Semin Pediatr Neurol, 2016; 23:127–133.
58.
BrainSpan Atlas of the Developing HumanBrain. Developmental transcriptomeATP1A3. www.brainspan.org/rnaseq/searches?exact_match=true&search_term=ATP1A3&search_type=gene&page_num=0 (accessed June16, 2020).
59.
ArystarkhovaE, HaqIU, LuebbertT, et al.Factors in the disease severity of ATP1A3 mutations: impairment, misfolding, and allele competition. Neurobiol Dis, 2019; 132:104577.
60.
PassiniMA, BuJ, RichardsAM, et al.Translational fidelity of intrathecal delivery of self-complementary AAV9-survival motor neuron 1 for spinal muscular atrophy. Hum Gene Ther, 2014; 25:619–630.
61.
HintonA, BondS, ForgacM. V-ATPase functions in normal and disease processes. Pflugers Arch, 2009; 457:589–598.
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