Novel erythropoietin-based therapeutic candidates with extra N-glycan sites that block hematopoiesis but preserve neuroplasticity

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

Biotechnology Journal

Wiley-WCH

Año: 2021

1860-6768

Neurological disorders affect millions of people worldwide causing behavior-cognitive disabilities. Spite of the effort does not exist any effective treatment for them nowadays. Human erythropoietin (EPO) has been used in vitro and in vivo studies because of its potential neurotropic and cytoprotective properties. However, its erythropoietic activity (EA) should be considered as a side effect. EPO-like molecules like asialoEPO, carbamylated-EPO or EPO-peptides have been developed to avoid hematological side effects during the treatment of neurobiological disorders. However, these molecules have different weaknesses: erythropoiesis preservation, low stability, potential immunogenicity, or fast clearance. Thus, we developed three EPO analogs using N-glycoengineering by adding a new glycosylation site within the EPO sequence responsible for its EA. In this work, we were cloned, produced and purified EPO-muteins, Mut 45_47, Mut 104, and Mut 151_153. All of them completely lost their EA in vitro and in vivo but preserved their neuroprotective action by rescuing staurosporine-induced apoptosis in hippocampal cultured neurons. Furthermore, the three EPO-muteins enhanced neuronal plasticity in hippocampal neurons more efficiently than hEPO. Interestingly, Mut 45_47 is a promising candidate to be explored as a biotherapeutic for neurological disorders considering not only its biological function but also its pharmacokinetic potential.