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

ORMAZABAL, MAXIMILIANO E.; VAENA, E.; PAVAN, E; MUCCI, J.M.; ROZENFELD P.; DARDIS, A

Liden

Simposio; IWGGD; 2022

BACKGROUND: Skeletal alterations in Gaucher patients arenot completely resolved by specific therapies. Moreover,bone mineral density pathophysiology is not completelyunderstood. Research studies revealed that both boneformation and resorption could be altered and play a role inbone pathology. Our group has developed a monocytemodel of Gaucher disease by specifically knocking-outGBA1 gene in THP-1 cell line, using CRISPR/Cas9 technology(THP-1 GBA1-KO). Edited cells showed reducedglucocerebrosidase (GCase) protein expression and andactivity, along with the increased level ofglucosylsphingosine (GlcSph).AIMS: We aim to further characterize the proinflammatorycytokines release and osteoclastogenesis in this GBA1 KOmodel and to study effect of different specific treatments.METHODS: THP1-wild type and GBA1-KO were grown inculture and exposed to the following treatments:recombinant human GCase (imiglucerase, ERT), eliglustat(substrate reduction therapy, SRT), ambroxol (chaperone)or Pentosan-polysulfate (PPS, anti-inflammatory). GCaseactivity, GlcSph, IL-1β and TNF-α levels and the tendency ofmonocytes to differentiate into osteoclasts were analyzed.RESULTS: THP-1 GBA1-KO cells displayed increased levels ofboth proinflammatory cytokines and osteoclastdifferentiation.GCase activity was detected only in cells treated with ERT.GlcSph accumulation was reduced, not only ERT and SRTtreatments, as expected, but also in cells treated withAmbroxol that promoted GlcSph release to the extracellularmedia. Levels of cytokines were modulated by all thetreatments at different levels. Osteoclastogenesis wasreduced by all the treatments.CONCLUSIONS: The THP-1 GCase KO model recapitulatesmost features of GD monocytes. All evaluated treatmentswere able to ameliorate, at least in part the pathologicalphenotype, as assessed by GlcSph levels, proinflammatorycytokines production and osteoclast differentiation.DISCLOSURE FOR ALL AUTHORS: PR received consultinghonoraria, travel and research grants from TakedaAD received consulting honoraria and travel grants fromTakeda and SanofiEP received a fellowship from Sanofi.