Dose-dependent treatment of optic nerve crush by exogenous systemic mutant erythropoietin.
The goal of the present study was to determine the minimum concentration of systemic erythropoietin-R76E required for neuroprotection in the retina. Erythropoietin (EPO) exhibits neuroprotective effects in both in vitro and in vivo models of neuronal cell death although its classical function is the regulation of red blood cell production. It can cross the blood brain barrier and therefore can be delivered systemically to affect the retina. However, long-term treatment with exogenous erythropoietin causes polycythemia. To decrease this potentially lethal effect, we generated and tested a modified form that contains a single arginine to glutamate mutation at the 76th position (EPO-R76E). In previous studies, this mutant protected retinal neurons in mouse models of retinal degeneration and glaucoma with similar efficacy as wild-type EPO. However, EPO-R76E has attenuated erythropoietic activity, therefore, neuroprotection can be achieved without causing a significant rise in hematocrit. BALB/cByJ mice received a single intramuscular injection of recombinant adeno-associated virus carrying enhanced green fluorescent protein, Epo, or Epo-R76E. To result in continuous production of four different doses of EPO-R76E, two doses of two different serotypes (2/5 and 2/8) were used. Mice were subjected to optic nerve crush and analysis was performed thirty days later. EPO-R76E showed dose-dependent protection of the retinal ganglion cell bodies, but was unable to prevent axonal degeneration. Furthermore, EPO-R76E induced a dose-dependent rise in the hematocrit that was still attenuated as compared to wild-type EPO.
Department of Ophthalmology, Hamilton Eye Institute, The University of Tennessee Health Science Center, Memphis, TN 38163, USA. timothy.sullivan@armstrong.edu
Timothy A
TA
Sullivan
Eldon E
EE
Geisert
Justin P
JP
Templeton
Tonia S
TS
Rex
D000818:Animals; D002452:Cell Count; D003587:Cytomegalovirus / Q000235:genetics; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D004921:Erythropoietin / Q000008:administration & dosage* / Q000235:genetics; D005822:Genetic Vectors; D005901:Glaucoma / Q000473:pathology / Q000517:prevention & control*; D049452:Green Fluorescent Proteins / Q000008:administration & dosage; D006400:Hematocrit; D007273:Injections, Intramuscular; D051379:Mice; D008807:Mice, Inbred BALB C; D009409:Nerve Crush; D020221:Optic Nerve Injuries / Q000473:pathology / Q000517:prevention & control*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage* / Q000235:genetics; D012165:Retinal Ganglion Cells / Q000187:drug effects* / Q000473:pathology
D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D004921:Erythropoietin; D049452:Green Fluorescent Proteins