P2.07. MODULATING MICROGLIAL AUTOPHAGY WITH IBRUTINIB: INSIGHTS FROM IN VITRO PHARMACOLOGY
Debanjan Das1, Akash S. Mali2, Denise Greco3, Danica Michaličková1, Jiří Novotný3, Ondřej Slanař1
1 Department of Pharmacology, First Faculty of Medicine, Charles University and General University Hospital Prague, Prague, Czech Republic
2 A. I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland
3 Department of Physiology, Faculty of Science, Charles University, Prague, Czech Republic
INTRODUCTION: Bruton’s tyrosine kinase (BTK) is key in B-cell signaling, autoimmunity, and inflammation, and may regulate autophagy. Ibrutinib, an FDA-approved BTK inhibitor, modulates immune cell function, including microglia. This project investigates how ibrutinib influences autophagy, oxidative stress, and neuroinflammation in glial cells, addressing a gap in neuroinflammatory disease research.
AIM(S): To investigate whether BTK inhibitor ibrutinib can show protective effects on mitochondria in glial cells in the LPS-induced model of neuroinflammation and explore the connection between mitochondria, BTK inhibition and autophagy.
METHOD(S): In-vitro cell treatment:
Murine microglial cell line (C8-B4) and newborn Wistar rats aged 1 to 5 days were sacrificed through decapitation. Primary mixed glial cells were obtained from the cortices and hippocampi of postnatal (P1–P2) rat pups and used for the in vitro cell culture.
Western blot
Western blot experiments performed as per standard protocol and multiple autophagy and inflammatory markers were used to detect targeted parameter.
RESULTS: Our study shows that ibrutinib modulates LPS-induced microglial activation by lowering TLR4 and NF-κB cytokine levels as well as activate autophagy . This suggests the involvement of the TLR4/NF-κB pathway Ibrutinib’s modulation of these pathways may help restore normal inflammatory responses.
Despite extensive research, inconsistencies remain in understanding microglial activation and polarization, and limited data exist on the role of ibrutinib or BTK inhibitors in microglial oxidative stress.
CONCLUSIONS: Therapeutic targeting of BTK by its inhibitors in inflammatory CNS disorders or treatment of autoimmune diseases is an emerging strategy that holds huge potential and can support current treatments of several neurological disorders and bring new paradigms toward brain disease.
FINANCIAL SUPPORT: This work was supported by Charles University project Cooperatio - Pharmaceutical Sciences and Charles University Grant Agency (GA UK) 162224.