PhD Scientific Days 2025

Poster Session I. - F: Pharmaceutical Sciences and Health Technologies

A New Therapeutic Approach in Heart Failure: A Cell-Based Platform for Testing Inflammasome Inhibitors

Előadó neve

Dr. Jezsoviczky Sára

Előadó munkahelye

Department of Pharmacology and Pharmacotherapy

Szerző(k) neve és munkahelye

Jezsoviczky Sára^1,2, Kovács Andrea^1,2,3, Kocsis Márton^1,2,3, Kozma Gergely⁴, Szabó Márta^1,2, Ferdinandy Péter^1,2,5,6, Varga Zoltán V.^1,2,3,7, Onódi Zsófia^1,2,3,7

1: Department of Pharmacology and Pharmacotherapy, Semmelweis University, H-1085 Budapest, Hungary
2: Center for Pharmacology and Drug Research & Development, Semmelweis University, H-1085 Budapest, Hungary
3: HCEMM-SU Cardiometabolic Immunology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, H-1089 Budapest, Hungary
4: Nanomedicine Research and Education Center, Semmelweis University, Budapest, Hungary
5: Pharmahungary Group, H-6722 Szeged, Hungary.
6: HUN-REN-SU System Pharmacology Research Group, Department of Pharmacology and Pharmacotherapy, Semmelweis University, H-1089 Budapest, Hungary
7: MTA-SE Momentum Cardio-Oncology and Cardioimmunology Research Group, Semmelweis University, Budapest, Hungary

Összefoglaló szövege

Introduction: Inflammasome activity is a key contributor in chronic inflammatory conditions, such as heart failure, so modulators of the NLRP3 inflammasome could be utilized in its therapy. The activation is a two-step process: priming, which upregulates the expression of inflammasome components, then activation, resulting in oligomerization of proteins and cytokine secretion (e.g., IL-1β). Discovering compounds that inhibit NLRP3 could lead to new methods to treat heart failure.
Aims: To develop a cell-based platform using a human monocytic cell line to evaluate potential inflammasome inhibitors.
Methods: THP1-ASC-GFP reporter cells were used to model inflammasome activation. NLRP3 inflammasome induction involved priming cells with lipopolysaccharide (LPS) for 24 hours, followed by activation with nigericin. Drug candidates — probenecid, sotagliflozin, canagliflozin, and linagliptin — were tested alongside known inhibitors MCC950 (NLRP3 inhibitor) and VX-765 (caspase-1 inhibitor). Cell viability was assessed to determine cell survival. Imaging flow cytometry was used to monitor inflammasome priming and activation, while Western blotting and caspase-1 activity assay were used to evaluate downstream signaling.
Results: Treatment with LPS and nigericin induced NLRP3 inflammasome activation, as demonstrated by increased ASC diffuse+ (18.34% to 55.67%) and ASC speck+ cell populations (0.97% to 49.40%), IL-1β cleavage and caspase-1 activity, accompanied by reduced cell viability. MCC950 suppressed NLRP3 activation (49.40% to 21.08%) and inhibited maturation of inflammatory mediators. In contrast, VX-765 had no effect on NLRP3 activation but reduced IL-1β cleavage (1.0 to 0.11) and caspase-1 activity. Among the tested compounds, canagliflozin and linagliptin significantly decreased cleaved IL-1β levels (to 0.45 and 0.53, respectively).
Conclusions: Our screening platform offers a tool to examine inflammasome activation and inhibition in THP1-ASC-GFP cells, supporting the discovery of new anti-inflammatory drugs. Our data suggests that approved drugs possess inflammasome modulatory effects, calling for further research into their mechanism of action.
Funding: The work was supported by János Bolyai Research Scholarship. Project no. RRF-2.3.1-21-2022-00003 has been implemented with the support provided by the European Union.