MONOCYTE IN VITRO MODEL OF GAUCHER DISEASE: EVALUATION OF THE EFFECT OF THERAPIES ON INFLAMMATION AND OSTEOCLASTOGENESIS

ORMAZABAL, MAXIMILIANO; VAENA, EMILIO; PAVAN, ELEONORA; MUCCI, JUAN M.; FERINO, DANIA; CIFÙ, ADRIANA; BIASIZZO, JESSICA; ROZENFELD, PAULA; DARDIS, ANDREA

Leiden

Simposio; 1st IWGGD Symposium 2022; 2022

BackgroundSkeletal alterations in Gaucher patients are not completely resolved by specific therapies. Moreover, bone mineral density pathophysiology is not completely understood. Research studies revealed that both bone formation and resorption could be altered and play a role in bone pathology. Our group has developed a monocyte model of Gaucher disease by specifically knocking-out GBA1 gene in THP-1 cell line, using CRISPR/Cas9 technology (THP-1 GBA1-KO). Edited cells showed reduced glucocerebrosidase (GCase) protein expression and and activity, along with the increased level of glucosylsphingosine (GlcSph).AimsWe aim to further characterize the proinflammatory cytokines release and osteoclastogenesis in this GBA1 KO model and to study effect of different specific treatments.MethodsTHP1-wild type and GBA1-KO were grown in culture and exposed to the following treatments: recombinant human GCase (imiglucerase, ERT), eliglustat (substrate reduction therapy, SRT), ambroxol (chaperone) or Pentosan-polysulfate (PPS, anti-inflammatory). GCase activity, GlcSph, IL-1β and TNF-α levels and the tendency of monocytes to differentiate into osteoclasts were analyzed.ResultsTHP-1 GBA1-KO cells displayed increased levels of both proinflammatory cytokines and osteoclast differentiation.GCase activity was detected only in cells treated with ERT. GlcSph accumulation was reduced , not only ERT and SRT treatments, as expected, but also in cells treated 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.ConclusionThe THP-1 GCase KO model recapitulates most features of GD monocytes. All evaluated treatments were able to ameliorate, at least in part the pathological phenotype, as assessed by GlcSph levels, proinflammatory cytokines production and osteoclast differentiation.