Monocyte in vitro model of Gaucher disease: Evaluation of the efect of therapies on inflammation and osteoclastogenesis

ORMAZABAL, MAXIMILIANO EMANUEL; VAENA, EMILIO; ELEONORA PAVAN; MUCCI, JUAN MARCOS; DANIA FERINO; ADRIANA CIFÙ

Leiden

Simposio; International Working Group on Gaucher Disease (IWGGD) 1st Symposium; 2023

IWGGD

Skeletal alterations in Gaucher patients are not completely resolved by specifictherapies. Moreover, bone mineral density pathophysiology is not completelyunderstood. Research studies revealed that both bone formation and resorptioncould be altered and play a role in bone pathology. Our group has developed amonocyte model of Gaucher disease by specifically knocking-out GBA1 gene inTHP-1 cell line, using CRISPR/Cas9 technology (THP-1 GBA1-KO). Edited cellsshowed reduced glucocerebrosidase (GCase) protein expression and and activity,along with the increased level of glucosylsphingosine (GlcSph).We aim to further characterize the proinflammatory cytokines release andosteoclastogenesis in this GBA1 KO model and to study effect of different specifictreatments.THP-1 GBA1-KO cells displayed increased levels of both proinflammatory cytokinesand osteoclast differentiation.GCase activity was detected only in cells treated with ERT. GlcSph accumulationwas reduced , not only ERT and SRT treatments, as expected, but also in cellstreated with Ambroxol that promoted GlcSph release to the extracellular media.Levels of cytokines were modulated by all the treatments at different levels.Osteoclastogenesis was reduced by all the treatments.The THP-1 GCase KO model recapitulates most features of GD monocytes. Allevaluated treatments were able to ameliorate, at least in part the pathologicalphenotype, as assessed by GlcSph levels, proinflammatory cytokines production andosteoclast differentiation.