NEW HYPERGLYCOSYLATED HUMAN ERYTHROPOIETIN-DERIVED MOLECULES AS THERAPEUTIC CANDIDATES FOR THE TREATMENT OF THE CENTRAL NERVOUS SYSTEM-AFFECTING DISEASES

BÜRGI, M ; APARICIO, G; KRATJE, RICARDO; SCORTICATI, C; OGGERO, M

Mar del Plata

Congreso; Reunión Anual de SOCIEDADES DE BIOCIENCIAS SAIC.SAFE.SAP.SAB 2019; 2019

SOCIEDADES DE BIOCIENCIAS SAIC.SAFE.SAP.SAB

Neurodegenerative diseases affect millions of people around the world causing cognitive and behaviour disorders. Nevertheless, does not exist any effective treatment for them nowadays. In this sense, human erythropoietin (EPO) has an important role considering its antiapoptotic, cytoprotective, angiogenic and antioxidant properties. However, its erythropoietic activity (EA) should be considered as a side effect. Thus, we proposed the development of new EPO analogues using an approach of N-glycoengineering by hyperglycosylation to preserve its neuroprotective and neuroplastic action but blocking the EA.New EPO muteins were obtained by adding one extra N-glycosylation site per molecule using site?directed mutagenesis. Then, they were produced in transduced CHO.K1 cells and purified by immunoaffinity chromatography. Primary cultures from hippocampal neurons were used to measure apoptosis inhibition, neuritogenesis, filopodia density and synapsis formation. In vitro and in vivo EA was also carried out.Three EPO variants were produced and one-step purified with a purity level higher than 89%. They presented an apparent molecular mass higher than EPO and a superior number of acidic isoforms as result of the increased glycosylation degree. The in vitro and the in vivo haematological activity of each EPO analogue was abolished (p