Neonatal encephalopathy due to perinatal hypoxia-ischemia (HI) is a severe condition, and current treatment options are limited. Expression of endogenous osteopontin (OPN), a multifunction glycoprotein, is strongly upregulated in the brain after neonatal HI. Intracerebrally administered OPN has been shown to be neuroprotective following experimental neonatal HI and adult stroke. In the present study, we determined whether intranasal, intraperitoneal or intracerebral treatment with a smaller TAT-OPN peptide is neuroprotective in neonatal mice with HI brain damage. The TAT-OPN peptide exerts bioactivity as it was as potent as full-length OPN in inducing cell adhesion in an in vitro adhesion assay. Intranasal administration of TAT-OPN peptide immediately after HI (T0) or in a repetitive treatment schedule of T0, 3 h, day (D) 1, 2 and 3 after HI did not protect cerebral gray or white matter after HI. Intraperitoneal TAT-OPN treatment at T0 or in two extended treatment schedules (D5, 7, 9, 11, 13, 15 after HI or T0, D1, 3, 5, 7, 9, 11, 13 and 15 after HI) did not result in neuroprotection either. Moreover, no functional improvement (cylinder rearing test and adhesive removal task) was observed following TAT-OPN treatment in any of the intraperitoneal treatment schedules. We validated that the TAT-OPN peptide reached the brain after intranasal or intraperitoneal administration by using an HIV-TAT staining. Finally, also intracerebral administration of the TAT-OPN peptide 1 h after HI did not reduce cerebral damage. Our data show that administration of the TAT-OPN peptide did not exert neuroprotective effects on neonatal HI-induced brain injury or sensorimotor behavioral deficits.

Armstrong-Wells J, Bernard TJ, Boada R, Manco-Johnson M: Neurocognitive outcomes following neonatal encephalopathy. Neurorehabilitation 2010;26:27-33.
De Haan M, Wyatt JS, Roth S, Vargha-Khadem F, Gadian D, Mishkin M: Brain and cognitive-behavioural development after asphyxia at term birth. Dev Sci 2006;9:350-358.
Glass HC, Hong KJ, Rogers EE, Jeremy RJ, Bonifacio SL, Sullivan JE, Barkovich AJ, Ferriero DM: Risk factors for epilepsy in children with neonatal encephalopathy. Pediatr Res 2011;70:535-540.
Gonzalez FF, Miller SP: Does perinatal asphyxia impair cognitive function without cerebral palsy? Arch Dis Child Fetal Neonatal Ed 2006;91:F454-F459.
Van Handel M, Swaab H, de Vries LS, Jongmans MJ: Long-term cognitive and behavioral consequences of neonatal encephalopathy following perinatal asphyxia: a review. Eur J Pediatr 2007;166:645-654.
Graham EM, Ruis KA, Hartman AL, Northington FJ, Fox HE: A systematic review of the role of intrapartum hypoxia-ischemia in the causation of neonatal encephalopathy. Am J Obstet Gynecol 2008;199:587-595.
Wyatt JS, Gluckman PD, Liu PY, Azzopardi D, Ballard R, Edwards AD, Ferriero DM, Polin RA, Robertson CM, Thoresen M, Whitelaw A, Gunn AJ; CoolCap Study Group: Determinants of outcomes after head cooling for neonatal encephalopathy. Pediatrics 2007;119:912-921.
Azzopardi DV, Strohm B, Edwards AD, Dyet L, Halliday HL, Juszczak E, Kapellou O, Levene M, Marlow N, Porter E, Thoresen M, Whitelaw A, Brocklehurst P; TOBY Study Group: Moderate hypothermia to treat perinatal asphyxial encephalopathy. N Engl J Med 2009;361:1349-1358.
Gluckman PD, Wyatt JS, Azzopardi D, Ballard R, Edwards AD, Ferriero DM, Polin RA, Robertson CM, Thoresen M, Whitelaw A, Gunn AJ: Selective head cooling with mild systemic hypothermia after neonatal encephalopathy: multicentre randomised trial. Lancet 2005;365:663-670.
Shankaran S, Laptook AR, Ehrenkranz RA, Tyson JE, McDonald SA, Donovan EF, Fanaroff AA, Poole WK, Wright LL, Higgins RD, Finer NN, Carlo WA, Duara S, Oh W, Cotten CM, Stevenson DK, Stoll BJ, Lemons JA, Guillet R, Jobe AH; National Institute of Child Health and Human Development Neonatal Research Network: whole-body hypothermia for neonates with hypoxic-ischemic encephalopathy. N Engl J Med 2005;353:1574-1584.
Bonestroo HJ, Nijboer CH, van Velthoven CT, Kavelaars A, Hack CE, van Bel F, Heijnen CJ: Cerebral and hepatic inflammatory response after neonatal hypoxia-ischemia in newborn rats. Dev Neurosci 2013;35:197-211.
Hedtjarn M, Mallard C, Eklind S, Gustafson-Brywe K, Hagberg H: Global gene expression in the immature brain after hypoxia-ischemia. J Cereb Blood Flow Metab 2004;24:1317-1332.
Hedtjarn M, Mallard C, Hagberg H: Inflammatory gene profiling in the developing mouse brain after hypoxia-ischemia. J Cereb Blood Flow Metab 2004;24:1333-1351.
Hagberg H, Gilland E, Bona E, Hanson LA, Hahin-Zoric M, Blennow M, Holst M, McRae A, Soder O: Enhanced expression of interleukin (IL)-1 and IL-6 messenger RNA and bioactive protein after hypoxia-ischemia in neonatal rats. Pediatr Res 1996;40:603-609.
Nijboer CH, Heijnen CJ, Groenendaal F, May MJ, van Bel F, Kavelaars A: A dual role of the NF-kappaB pathway in neonatal hypoxic-ischemic brain damage. Stroke 2008;39:2578-2586.
Van Velthoven CT, Heijnen CJ, van Bel F, Kavelaars A: Osteopontin enhances endogenous repair after neonatal hypoxic-ischemic brain injury. Stroke 2011;42:2294-2301.
Vexler ZS, Ferriero DM: Molecular and biochemical mechanisms of perinatal brain injury. Semin Neonatol 2001;6:99-108.
O'Regan A, Berman JS: Osteopontin: a key cytokine in cell-mediated and granulomatous inflammation. Int J Exp Pathol 2000;81:373-390.
Chen W, Ma Q, Suzuki H, Hartman R, Tang J, Zhang JH: Osteopontin reduced hypoxia-ischemia neonatal brain injury by suppression of apoptosis in a rat pup model. Stroke 2011;42:764-769.
Chakraborty G, Jain S, Behera R, Ahmed M, Sharma P, Kumar V, Kundu GC: The multifaceted roles of osteopontin in cell signaling, tumor progression and angiogenesis. Curr Mol Med 2006;6:819-830.
Denhardt DT, Noda M, O'Regan AW, Pavlin D, Berman JS: Osteopontin as a means to cope with environmental insults: regulation of inflammation, tissue remodeling, and cell survival. J Clin Invest 2001;107:1055-1061.
Mazzali M, Kipari T, Ophascharoensuk V, Wesson JA, Johnson R, Hughes J: Osteopontin - a molecule for all seasons. Q J Med 2002;95:3-13.
Doyle KP, Yang T, Lessov NS, Ciesielski TM, Stevens SL, Simon RP, King JS, Stenzel-Poore MP: Nasal administration of osteopontin peptide mimetics confers neuroprotection in stroke. J Cereb Blood Flow Metab 2008;28:1235-1248.
Meller R, Stevens SL, Minami M, Cameron JA, King S, Rosenzweig H, Doyle K, Lessov NS, Simon RP, Stenzel-Poore MP: Neuroprotection by osteopontin in stroke. J Cereb Blood Flow Metab 2005;25:217-225.
Suzuki H, Ayer R, Sugawara T, Chen W, Sozen T, Hasegawa Y, Kanamaru K, Zhang JH: Protective effects of recombinant osteopontin on early brain injury after subarachnoid hemorrhage in rats. Crit Care Med 2010;38:612-618.
Suzuki H, Hasegawa Y, Kanamaru K, Zhang JH: Mechanisms of osteopontin-induced stabilization of blood-brain barrier disruption after subarachnoid hemorrhage in rats. Stroke 2010;41:1783-1790.
Nijboer CH, Bonestroo HJ, Zijlstra J, Kavelaars A, Heijnen CJ: Mitochondrial JNK phosphorylation as a novel therapeutic target to inhibit neuroinflammation and apoptosis after neonatal ischemic brain damage. Neurobiol Dis 2013;54:432-444.
Nijboer CH, Heijnen CJ, Groenendaal F, May MJ, van Bel F, Kavelaars A: Strong neuroprotection by inhibition of NF-kappaB after neonatal hypoxia-ischemia involves apoptotic mechanisms but is independent of cytokines. Stroke 2008;39:2129-2137.
Yang D, Sun YY, Lin X, Baumann JM, Dunn RS, Lindquist DM, Kuan CY: Intranasal delivery of cell-penetrating anti-NF-kappaB peptides (Tat-NBD) alleviates infection-sensitized hypoxic-ischemic brain injury. Exp Neurol 2013;247:447-455.
Yin W, Cao G, Johnnides MJ, Signore AP, Luo Y, Hickey RW, Chen J: TAT-mediated delivery of Bcl-xL protein is neuroprotective against neonatal hypoxic-ischemic brain injury via inhibition of caspases and AIF. Neurobiol Dis 2006;21:358-371.
Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C: A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nat Med 2003;9:1180-1186.
Ellison JA, Velier JJ, Spera P, Jonak ZL, Wang X, Barone FC, Feuerstein GZ: Osteopontin and its integrin receptor alpha(v)beta3 are upregulated during formation of the glial scar after focal stroke. Stroke 1998;29:1698-1706; discussion 1707.
Lee MY, Shin SL, Choi YS, Kim EJ, Cha JH, Chun MH, Lee SB, Kim SY: Transient upregulation of osteopontin mRNA in hippocampus and striatum following global forebrain ischemia in rats. Neurosci Lett 1999;271:81-84.
Wang X, Louden C, Yue TL, Ellison JA, Barone FC, Solleveld HA, Feuerstein GZ: Delayed expression of osteopontin after focal stroke in the rat. J Neurosci 1998;18:2075-2083.
Noiri E, Dickman K, Miller F, Romanov G, Romanov VI, Shaw R, Chambers AF, Rittling SR, Denhardt DT, Goligorsky MS: Reduced tolerance to acute renal ischemia in mice with a targeted disruption of the osteopontin gene. Kidney Int 1999;56:74-82.
Hashimoto M, Sun D, Rittling SR, Denhardt DT, Young W: Osteopontin-deficient mice exhibit less inflammation, greater tissue damage, and impaired locomotor recovery from spinal cord injury compared with wild-type controls. J Neurosci 2007;27:3603-3611.
Liaw L, Birk DE, Ballas CB, Whitsitt JS, Davidson JM, Hogan BL: Altered wound healing in mice lacking a functional osteopontin gene (spp1). J Clin Invest 1998;101:1468-1478.
Schroeter M, Zickler P, Denhardt DT, Hartung HP, Jander S: Increased thalamic neurodegeneration following ischaemic cortical stroke in osteopontin-deficient mice. Brain 2006;129:1426-1437.
DeBow SB, Clark DL, MacLellan CL, Colbourne F: Incomplete assessment of experimental cytoprotectants in rodent ischemia studies. Can J Neurol Sci 2003;30:368-374.
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