A partially-folded hyperglycosylated erythropoietin with no erythropoietic activity which protects neurons from damage and promotes neuroplasticity

BURGI, M. M.; SCORTICATI, C.; VAGLIENTI, V.; SÁNCHEZ, M. C.; DEPETRIS, M.; KRATJE, R. B.; OGGERO, M.

Viña del Mar. Valparaíso

Simposio; X Simposio Latinoamericano de Tecnología de Cultivo de Células Animales (X SLATCC); 2024

Pontificia Universidad Católica de Valparaíso. Universidad de Chile. Universidad de Concepción.

Background: Neurodegenerative diseases (ND) are characterized by their chronicity andprogressive evolution impacting significantly in the quality of life of patients. These diseasescompromise the viability and functionality of neurons. Despite their increasing incidence, effectivetreatments are often lacking or insufficient. Erythropoietin (EPO) is a promising candidate due to itsneurobiological actions (NA); however, its erythropoietic activity (EA) causes undesirable effects.This study aimed to describe an innovative EPO-derived neurotherapeutic candidate. It wasobtained through glycoengenieering hEPO by hyperglycosylation to block its EA while preservingits NA.Methods: A glycoengineered EPO variant, named BSY12, was developed. BSY12 was produced inCHO.K1 cells, immunoaffinity purified, and extensively characterized. The protein´s structure andstability were assessed using circular dichroism and thermal shift analysis. EA was evaluated bymeasuring hematocrit in normocytemic BALB/c mice. The BSY12 antiapoptotic and neuroplasticactivities were studied in primary hippocampal neuron cultures. Additionally, Sholl analysis wasperformed on CA1 pyramidal neurons from BSY12-treated CF1 mice. The ability to cross theblood-brain barrier (BBB) was investigated in CF1, C57BI/6N, and SOD1G93A mice. In vivoneurobiological performance was evaluated using the Complex Running Wheels test in healthyC57BI/6N mice and in an amyotrophic lateral sclerosis model with SOD1G93A mice. Finally, itsneuroprotective and antigliotic effects were studied in Müller cells (retinal ganglion cells).Results: BSY12 exhibited a partially-folded structure in comparison with hEPO; nevertheless, theEA was blocked while its NA was preserved. BSY12 reduced staurosporine-induced apoptosis in vitro in hippocampal neurons and promoted neurite outgrowth, the augment of filopodia density,and synapse formation (p