Intranasal formulation of erythropoietin (EPO) showed potent protective activity against amyloid toxicity in the Aβ 25-35 non-transgenic mouse model of Alzheimer’s disease
Restricted accessResearch articleFirst published online November, 2013
Intranasal formulation of erythropoietin (EPO) showed potent protective activity against amyloid toxicity in the Aβ 25-35 non-transgenic mouse model of Alzheimer’s disease
Erythropoietin (EPO) promotes neurogenesis and neuroprotection. We here compared the protection induced by two EPO formulations in a rodent model of Alzheimer’s disease (AD): rHu-EPO and a low sialic form, Neuro-EPO. We used the intracerebroventricular administration of aggregated Aβ25-35 peptide, a non-transgenic AD model. rHu-EPO was tested at 125–500 µg/kg intraperitoneally and Neuro-EPO at 62–250 µg/kg intranasally (IN). Behavioural procedures included spontaneous alternation, passive avoidance, water-maze and object recognition, to address spatial and non-spatial, short- and long-term memories. Biochemical markers of Aβ25-35 toxicity in the mouse hippocampus were examined and cell loss in the CA1 layer was determined. rHu-EPO and Neuro-EPO led to a significant prevention of Aβ25-35-induced learning deficits. Both EPO formulations prevented the induction of lipid peroxidation in the hippocampus, showing an antioxidant activity. rHu-EPO (250 µg/kg) or Neuro-EPO (125 µg/kg) prevented the Aβ25-35-induced increase in Bax level, TNFα and IL-1β production and decrease in Akt activation. A significant prevention of the Aβ25-35-induced cell loss in CA1 was also observed. EPO is neuroprotective in the Aβ25-35 AD model, confirming its potential as an endogenous neuroprotection system that could be boosted for therapeutic efficacy. We here identified a new IN formulation of EPO showing high neuroprotective activity. Considering its efficacy, ease and safety, IN Neuro-EPO is a new promising therapeutic agent in AD.
AdamcioBSarginDStradomskaA, et al. (2008) Erythropoietin enhances hippocampal long-term potentiation and memory. BMC Biol6: 37.
2.
AssarafMIDiazZLibermanA, et al. (2007) Brain erythropoietin receptor expression in Alzheimer disease and mild cognitive impairment. J Neuropathol Exp Neurol66: 389–398.
3.
BartesaghiSMarinovichMCorsiniE, et al. (2005) Erythropoietin: A novel neuroprotective cytokine. Neurotoxicology26: 923–928.
4.
BernaudinMMartiHHRousselS, et al. (1999) A potential role for erythropoietin in focal permanent cerebral ischemia in mice. J Cereb Blood Flow Metab19: 643–651.
5.
BhatRVShanleyJCorrellMP, et al. (2000) Regulation and localization of tyrosine216 phosphorylation of glycogen synthase kinase-3β in cellular and animal models of neuronal degeneration. Proc Natl Acad Sci USA97:11074–11079.
6.
BrinesMLGhezziPKeenanS, et al. (2000) Erythropoietin crosses the blood-brain barrier to protect against experimental brain injury. Proc Natl Acad Sci USA97: 10526–10531.
7.
BrunetABonniAZigmondMJ, et al. (1999) Akt promotes cell survival by phosphorylating and inhibiting a Forkhead transcription factor. Cell96: 857–868.
8.
CelikMGökmenNErbayraktarS, et al. (2002) Erythropoietin prevents motor neuron apoptosis and neurologic disability in experimental spinal cord ischemic injury. Proc Natl Acad Sci USA99: 2258–2263.
9.
ChavantFDeguilJPainS, et al. (2010) Imipramine, in part through tumor necrosis factor α inhibition, prevents cognitive decline and β-amyloid accumulation in a mouse model of Alzheimer’s disease. J Pharmacol Exp Ther332: 505–514.
10.
ChengIHScearce-LevieKLegleiterJ, et al. (2007) Accelerating amyloid-β fibrillization reduces oligomer levels and functional deficits in Alzheimer disease mouse models. J Biol Chem282: 23818–23828.
11.
ChinKYuXBeleslin-CokicB, et al. (2000) Production and processing of erythropoietin receptor transcripts in brain. Brain Res Mol Brain Res81: 29–42.
12.
ChongZZKangJKMaieseK (2003b) Apaf-1, Bcl-xL, cytochrome c, and caspase-9 Form the critical elements for cerebral vascular protection by erythropoietin. J Cereb Blood Flow Metab23: 320–330.
13.
ChongZZKangJKMaieseK (2004) Akt1 drives endothelial cell membrane asymmetry and microglial activation through Bcl-x(L) and caspase 1, 3, and 9. Exp Cell Res296: 196–207.
14.
ChongZZLiFMaieseK (2005) Erythropoietin requires NF-κB and its nuclear translocation to prevent early and late apoptotic neuronal injury during β-amyloid toxicity. Curr Neurovasc Res2: 387–399.
15.
ChongZZLinSHKangJK, et al. (2003a) Erythropoietin prevents early and late neuronal demise through modulation of Akt1 and induction of caspase 1, 3, and 8. J Neurosci Res71: 659–669.
16.
CraftSBakerLDMontineTJ, et al. (2012) Intranasal insulin therapy for Alzheimer disease and amnestic mild cognitive impairment: A pilot clinical trial. Arch Neurol69: 29–38.
17.
DelobetteSPrivatAMauriceT (1997) In vitro aggregation facilities β-amyloid peptide- (25–35)-induced amnesia in the rat. Eur J Pharmacol319: 1–4.
18.
DigicayliogluMBichetSMartiHH, et al. (1995) Localization of specific erythropoietin binding-sites in defined areas of the mouse-brain. Proc Natl Acad Sci USA92: 3717–3720.
19.
FletcherLKohliSSpragueSM, et al. (2009) Intranasal delivery of erythropoietin plus insulin-like growth factor-I for acute neuroprotection in stroke. J Neurosurg111: 164–170.
20.
Garcia-RodríguezJCRodríguez-CruzY (2012) The therapeutic potential of Neuro-EPO administered nasally on acute cerebrovascular disease. Curr Psychopharmacol1: 228–232.
21.
Garcia-RodríguezJCSosa-TestéI (2009) The nasal route as a potential pathway for delivery of erythropoietin in the treatment of acute ischemic stroke in humans. Sci World J9: 970–981.
22.
GencSZadeoglulariZOnerMG, et al. (2011) Intranasal erythropoietin therapy in nervous system disorders. Expert Opin Drug Deliv8: 19–32.
23.
GrunfeldJFBarhumYBlondheimN, et al. (2007) Erythropoietin delays disease onset in an amyotrophic lateral sclerosis model. Exp Neurol204: 260–263.
24.
HooperCKillickRLovestoneS (2008) The GSK-3 hypothesis of Alzheimer’s disease. J Neurochem104: 1433–1439.
25.
Hoynck van PapendrechtMAJeekelHBuschOR, et al. (1992) Efficacy of recombinant erythropoietin for stimulating erythropoiesis after blood loss and surgery. An experimental study in rats. Eur J Surg158: 83–87.
26.
HsiaoKChapmanPNilsenS, et al. (1996) Correlative memory deficits, Aβ elevation, and amyloid plaques in transgenic mice. Science274: 99–102.
27.
JuulSEYachnisATChristensenRD (1998) Tissue distribution of erythropoietin and erythropoietin receptor in the developing human fetus. Early Hum Dev52: 235–249.
28.
KamalAAl ShaibaniTRamakersG (2011) Erythropoietin decreases the excitatory neurotransmitter release probability and enhances synaptic plasticity in mice hippocampal slices. Brain Res1410: 33–37.
29.
KawakamiMSekiguchiMSatoK, et al. (2001) Erythropoietin receptor-mediated inhibition of exocytotic glutamate release confers neuroprotection during chemical ischemia. J Biol Chem276: 39469–39475.
30.
KlementievBNovikovaTNovitskayaV, et al. (2007) A neural cell adhesion molecule-derived peptide reduces neuropathological signs and cognitive impairment induced by Aβ25–35. Neuroscience145: 209–224.
31.
KlyubinIWalshDMLemereCA, et al. (2005) Amyloid-β protein immunotherapy neutralizes Aβ oligomers that disrupt synaptic plasticity in vivo. Nat Med11: 556–561.
KumralAUysalNTugyanK, et al. (2004) Erythropoietin improves long-term spatial memory deficits and brain injury following neonatal hypoxia-ischemia in rats. Behav Brain Res153: 77–86.
34.
LeeSTChuKParkJE, et al. (2012) Erythropoietin improves memory function with reducing endothelial dysfunction and amyloid-β burden in Alzheimer’s disease models. J Neurochem120: 115–124.
35.
LeeSTChuKSinnDI, et al. (2006) Erythropoietin reduces perihematomal inflammation and cell death with eNOS and STAT3 activations in experimental intracerebral hemorrhage. J Neurochem96: 1728–1739.
36.
LiGMaRHuangC, et al. (2008) Protective effect of erythropoietin on β-amyloid-induced PC12 cell death through antioxidant mechanisms. Neurosci Lett442: 143–147.
37.
LiWMaedaYYuanRR, et al. (2004) Beneficial effect of erythropoietin on experimental allergic encephalomyelitis. Ann Neurol56: 767–777.
38.
LueLFKuoYMRoherAE, et al. (1999) Soluble amyloid β peptide concentration as a predictor of synaptic change in Alzheimer’s disease. Amer J Pathol155: 853–862.
39.
MaRXiongNHuangC, et al. (2009) Erythropoietin protects PC12 cells from β-amyloid25–35-induced apoptosis via PI3K/Akt signaling pathway. Neuropharmacology56: 1027–1034.
40.
MaieseKChongZZLiF, et al. (2008) Erythropoietin: Elucidating new cellular targets that broaden therapeutic strategies. Prog Neurobiol85: 194–213.
41.
MaieseKLiFChongZZ (2004) Erythropoietin in the brain: can the promise to protect be fulfilled?Trends Pharmacol Sci25: 577–583.
42.
MajumdarACruzDAsamoahN, et al. (2007) Activation of microglia acidifies lysosomes and leads to degradation of Alzheimer amyloid fibrils. Mol Biol Cell18: 1490–1496.
43.
MartínDSalinasMLópez-ValdalisoR, et al. (2001) Effect of the Alzheimer amyloid fragment Aβ25–35 on Akt/PKB kinase and survival of PC12 cells. J Neurochem78: 1000–1008.
44.
MattsonMP (2004) Pathways towards and away from Alzheimer’s disease. Nature430: 631–639.
45.
MauriceTLockhartBPPrivatA (1996) Amnesia induced in mice by centrally administered β-amyloid peptides involves cholinergic dysfunction. Brain Res706: 181–193.
46.
McLeanCAChernyRAFraserFW, et al. (1999) Soluble pool of Aβ amyloid as a determinant of severity of neurodegeneration in Alzheimer’s disease. Ann Neurol46: 860–866.
47.
MeunierJIeniJMauriceT (2006) The anti-amnesic and neuroprotective effects of donepezil against amyloid β25–35 peptide-induced toxicity in mice involve an interaction with the σ1 receptor. Br J Pharmacol149: 998–1012.
48.
MitrasinovicOMMurphyGM (2003) Microglial overexpression of the M-CSF receptor augments phagocytosis of opsonized Abeta. Neurobiol Aging24: 807–815.
49.
MitrasinovicOMVincentVASimsekD, et al. (2003) Macrophage colony stimulating factor promotes phagocytosis by murine microglia. Neurosci Lett344: 185–188.
50.
MorishitaEMasudaSNagaoM, et al. (1997) Erythropoietin receptor is expressed in rat hippocampal and cerebral cortical neurons, and erythropoietin prevents in vitro glutamate-induced neuronal death. Neuroscience76: 105–116.
51.
NadamJNavarroFSanchezP, et al. (2007) Neuroprotective effects of erythropoietin in the rat hippocampus after pilocarpine-induced status epilepticus. Neurobiol Dis25: 412–426.
52.
NaertGRivestS (2011) The role of microglial cell subsets in Alzheimer’s disease. Curr Alzheimer Res8: 151–155.
53.
NagaiANakagawaEChoiHB, et al. (2001) Erythropoietin and erythropoietin receptors in human CNS neurons, astrocytes, microglia, and oligodendrocytes grown in culture. J Neuropathol Exp Neurol60: 386–392.
54.
ParadisEDouillardHKoutroumanisM, et al. (1996) Amyloid β peptide of Alzheimer’s disease downregulates Bcl-2 and upregulates bax expression in human neurons. J Neurosci16: 7533–7539.
55.
PaxinosGFranklinKBJ (2004) The mouse brain in stereotaxic coordinates. San Diego, USA: Elsevier Academic Press.
56.
RabieTMartiHH (2008) Brain protection by erythropoietin: a manifold task. Physiology (Bethesda)23: 263–274.
57.
Rodríguez CruzYMengana TámosYMuñoz CernudaA, et al. (2010) Treatment with nasal neuro-EPO improves the neurological, cognitive, and histological state in a gerbil model of focal ischemia. Sci World J10: 2288–2300.
58.
SakanakaMWenTCMatsudaS, et al. (1998) In vivo evidence that erythropoietin protects neurons from ischemic damage. Proc Natl Acad Sci USA95: 4635–4640.
59.
SanchezPEFaresRPRissoJJ, et al. (2009) Optimal neuroprotection by erythropoietin requires elevated expression of its receptor in neurons. Proc Natl Acad Sci USA106: 9848–9853.
60.
ScopesDICO’HareEJeggoR, et al. (2012) Aβ oligomer toxicity inhibitor protects memory in models of synaptic toxicity. Br J Pharmacol167: 383–392.
SomervailleTCLinchDCKhwajaA (2001) Growth factor withdrawal from primary human erythroid progenitors induces apoptosis through a pathway involving glycogen synthase kinase-3 and Bax. Blood98: 1374–1381.
63.
Sosa TestéIGarcía RodríguezJCGarcía SalmanJD, et al. (2006) Intranasal administration of recombinant human erythropoietin exerts neuroprotective effects on post-ischemic brain injury in mongolian gerbils. Pharmacol Online1: 100–112.
64.
StepanichevMYMoiseevaYVLazarevaNA, et al. (2003) Single intracerebroventricular administration of amyloid-β25–35 peptide induces impairment in short-term rather than long-term memory in rats. Brain Res Bull61: 197–205.
65.
StepanichevMYZdobnovaIMZarubenkoII, et al. (2004) Amyloid-β25–35-induced memory impairments correlate with cell loss in rat hippocampus. Physiol Behav80: 647–655.
66.
StepanichevMYZdobnovaIMZarubenkoII, et al. (2006) Studies of the effects of central administration of β-amyloid peptide (25–35): Pathomorphological changes in the hippocampus and impairment of spatial memory. Neurosci Behav Physiol36: 101–106.
67.
VillardVEspallerguesJKellerE, et al. (2009) Antiamnesic and neuroprotective effects of the aminotetrahydrofuran derivative ANAVEX1–41 against amyloid β25–35-induced toxicity in mice. Neuropsychopharmacology34: 1552–1566.
68.
VillardVEspallerguesJKellerE, et al. (2011) Anti-amnesic and neuroprotective potentials of the mixed muscarinic receptor/sigma1 (σ1) ligand ANAVEX2–73, a novel aminotetrahydrofuran derivative. J Psychopharmacol25: 1101–1117.
69.
WalshDMSelkoeDJ (2007) Aβ oligomers – a decade of discovery. J Neurochem101: 1172–1184.
ZussyCBrureauADelairB, et al. (2011) Time-course and regional analyses of the physiopathological changes induced after cerebral injection of an amyloid β fragment in rats. Am J Pathol179: 315–334.
