PhD Scientific Days 2023

Budapest, 22-23 June 2023

Translational Medicine - Posters O

Role of Inflammasome Activation in Monocytes with Hypercholesterolemia

Regina N. Nagy1, Zsófia Gulyás-Onódi1,2, Péter Ferdinandy1, Anikó Görbe1, Zoltán V. Varga1,2
1 Department of Pharmacology and Pharmacotherapy, Semmelweis University, Budapest
2 HCEMM-SU Cardiometabolic Immunology Research Group, Budapest

Text of the abstract

Introduction: Inflammation plays a critical role in the genesis and progression of cardiovascular diseases. Inflammasomes are important players in inflammatory processes. NLRP3 inflammasome activation is engaged in several cardiovascular and metabolic diseases. Cholesterol, a known cardiovascular metabolic risk factor, was shown to activate NLRP3 inflammasome in macrophages, however, little is known about AIM2 or NLRC4 inflammasome activation in hypercholesterolemic pathologies. Direct inhibition of inflammasomes, rather than inflammatory cytokine suppression, can be an effective anti-inflammatory management. Thus, exploring the molecular mechanisms of inflammasome activation can lead to the discovery of new potential drug targets in hypercholesterolemic cardiovascular comorbidities.
Aims: To investigate the effect of in vitro hypercholesterolemic treatment on the inflammatory mechanisms of THP1-ASC-GFP monocytes.
Methods: Cells were treated with hypercholesterolemic supplements for 72 hours in 3 different concentrations (AAL1, AAL2, AAL3) according to our previously set cell culture model, or water soluble cholesterol (20, 40, 80, 160 µg/mL), or ox-LDL (12.5, 25, 50 µg/mL), then cell viability and cholesterol uptake were measured. After 72 hours of AAL2 treatment, NLRP3, NLRC4 and AIM2 inflammasomes were stimulated with LPS+Nigericin, Flagellin and Poly(dA:dT), respectively for 24 hours, and inflammasome marker protein (NLRP3, NLRC4, AIM2) expression was tested by Western blot.
Result: Treatment of THP1 monocytes with 25 or 50 µg/mL ox-LDL for 72 hours significantly reduced cell viability, while other hypercholesterolemic treatments did not affect it. All treatments resulted elevated intracellular cholesterol level. 72 hours of hypercholesterolemic treatment did not affect NLRP3, AIM2 or NLRC4 protein expression in either setup.
Conclusion: Hypercholesterolemia did not affect inflammasome marker protein expression in THP1-ASC-GFP monocytes in present in vitro system.
Funding: The work was supported by the European Union's Horizon 2020 research and innovation programme under grant agreement No 739593. Project no. TKP2021-EGA-23 has been implemented with the support provided by the Ministry of Innovation and Technology of Hungary from the National Research, Development and Innovation Fund, financed under the TKP2021-EGA funding scheme. RRF-2.3.1-21-2022-00003.