PB0803 - P2Y12 receptor expression on monocytes, monocyte-derived macrophages and dendritic cells and modulation by ticagrelor

Monday, June 26, 2023

18:30 – 19:30 ET

Room: Exhibition

Co-Author(s)

Adeline PONTIS, MD

Department of Hematology/Hemostasis, Pontchaillou University Hospital of Rennes, Rennes, France ; Univ Rennes, CHU Rennes, Inserm, EHESP, Irset (Institut De Recherche En Santé, Environnement Et Travail) - UMR_S 1085, Rennes, France
RENNES, Bretagne, France

Co-Author(s)

AL

Alain LESCOAT

MD, PhD
Department of Internal Medicine and Clinical Immunology, Rennes University Hospital, 35203 Rennes, France ; Univ Rennes, CHU Rennes, Inserm, EHESP, Irset (Institut De Recherche En Santé, Environnement Et Travail) - UMR_S 1085, Rennes, France
Rennes, Bretagne, France
ML

Marie LELONG

Technician
Univ Rennes, CHU Rennes, Inserm, EHESP, Irset (Institut De Recherche En Santé, Environnement Et Travail) - UMR_S 1085, Rennes, France.
Rennes, Bretagne, France
OF

Olivier FARDEL

PharmD, PhD
Univ Rennes, CHU Rennes, Inserm, EHESP, Irset (Institut De Recherche En Santé, Environnement Et Travail) - UMR_S 1085, Rennes, France ; Department of Hematology, Rennes University Hospital, 35203 Rennes, France
Rennes, Bretagne, France

Presenting Author(s)

Isabelle Gouin-Thibault, Sr., PhD

Head of hemostasis department
Department of Hematology/Hemostasis, Pontchaillou University Hospital of Rennes, Univ Rennes, Irset - Inserm UMR_S 1085, Rennes, France.
Rennes, Bretagne, France

Co-Author(s)

VL

Valérie LECUREUR

PhD
Univ Rennes, CHU Rennes, Inserm, EHESP, Irset (Institut De Recherche En Santé, Environnement Et Travail) - UMR_S 1085, Rennes, France.
Rennes, Bretagne, France

Background: P2Y12, a transmembrane receptor to ADP mainly expressed on platelets, is also found on immune cells, suggesting a role in inflammation. However, its profile of expression on these cells and the modulation by P2Y12R inhibitors are poorly understood.

Aims: To characterize the expression of P2Y12R on monocytes, monocyte-derived macrophages (MDM) and dendritic cells (DC) and its modulation by ticagrelor.

Methods: Peripheral blood mononuclear cells were collected from buffy coats (healthy donors, French Blood Establishment). Monocytes were differentiated into MDMs or DCs (M-CSF of GM-CSF+IL-4 respectively) during 6 days. MDMs were then polarized for 24 or 48 hours into MDM-M1 (LPS+IFNγ), MDM-M2a (IL-4+IL-13), MDM-M2c (IL-10) or MDM-M2dex (IL-10+dexamethasone) and DCs into immature (DCi) or mature (DCm, by addition of LPS for 24h), in presence or not of ticagrelor (1.5 µM) in the culture medium.
After mRNA extraction (TRIzol™) and reverse transcription, quantitative PCRs were performed.
P2Y12R expression was determined using flow cytometric direct immunofluorescence (FC) with the PE-anti-P2Y12 antibody.

Results: P2Y12R mRNA expression was found similar in monocytes and in MDMs whereas its expression was significantly induced during the differentiation of monocytes into DCi (mean fold change=9.3+/-8.8 versus monocytes) but repressed in DCm versus DCi (mean fold change=0.30+/-0.24), showing a suppressive effect of LPS (Figure 1).
During MDM polarization, P2Y12R mRNA expression was repressed in MDM-M1 and strongly induced in MDM-M2a (mean fold change=16.1+/-6.8) and MDM-M2dex (mean fold change=24.5+/-12.5), in favor of an enhancing effect of glucocorticoids (Figure 2). The P2Y12R expression profile in MDM-M1 and MDM-M2a was confirmed by FC after cell permeabilization.
In the presence of ticagrelor, P2Y12R mRNA expression showed a tendency to decrease in MDM-M2a (p=0.06).

Conclusion(s): This original work revealed that P2Y12R expression was different according to the MDM-polarization profile. Ticagrelor did not show a significant direct effect on P2Y12R expression, further studies are ongoing to investigate its indirect effect.