Hyperglycosylated Epo Variants To Treat Neurodegenerative Diseases

BURGI, MARIA DE MILAGROS ; CAMILA SCORTICATI; AQUILES DORELLA; KRATJE, RICARDO; APARICIO GABRIELA; OGGERO, MARCOS

copenaghem

Congreso; Esact 2019; 2019

esact2019

Background and novelty: Neurodegenerative diseases affect the nervous system causing cognitive and behavior disorders. Sincethere is no effective therapy, new neuroprotective and neurotrophic agents are necessary. In this sense, human erythropoietin (EPO)has an important role considering its antiapoptotic, cytoprotective, angiogenic and antioxidant properties. Nevertheless, itshematological activity (HA) should be considered as a side effect. EPO molecular regions responsible of neuroprotective orneuroplastic activity are dissociated from those linked to HA. Thus, the incorporation of new potential N-glycosylation sites wascarried out in EPO regions identified as essential for displaying the HA avoiding modification of residues needed to retain theneuropropertiesExperimental approach: EPO analogs were obtained by adding new extra N-glycosylation sites by site?directed mutagenesis andproduced in transduced CHO.K1 cells. Muteins were purified by immunoafinity chromatography. Primary cultures from hippocampalneurons were used to measure apoptosis inhibition, neuritogenesis, filopodia density and synapsis formation. In vitro and in vivo HAassays were also carried outResults and discussion: Three EPO variants were produced and purified with a purity level higher than 89% in only one step.They presented a molecular weight higher than EPO and a superior number of acid isoforms as result of the increased glycosylationdegree. None of the variants evidenced HA evaluated both in vitro and in vivo (p