OC 61.2 - MiR-146a-mimic therapy modulates platelet reactivity preventing mortality caused by pulmonary embolism model

Tuesday, June 27, 2023

15:00 – 15:15 ET

Room: 710A

Session Available As: On-Demand

Presenting Author(s)

Sonia Aguila, PhD (she/her/hers)

Researcher
Centro Regional de Hemodonación, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain

Co-Author(s)

JG

Juan carlos Garcia-Cañaveras

Postdoc
Biomarkers and Precision Medicine Unit, Instituto de Investigación Sanitaria Fundación Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
Ascension M M. De los Reyes-Garcia, MSc

PhD student
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
LZ

Laura Zapata-Martinez

PhD student
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
NG

Nuria Garcia-Barbera

Technician
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
PG

Pedro J Guijarro

Technician
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
ML

María Luisa Lozano

Leader of Hematology Service in Murcia
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, CIBERER, Murcia, Spain
Murcia, Murcia, Spain
BP

Bernard Payrastre

Professor, MD
INSERM, UMR-S U1297 and University of Toulouse III, Institute of Cardiovascular and Metabolic Diseases (I2MC), CHU-Rangueil, laboratoire d'Hématologie, CHU de Toulouse, Toulouse, France
Toulouse, Languedoc-Roussillon, France
José RIVERA Rivera, PhD

PhD
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
MG

Marie Pierre Gratacap

Professor, PhD
INSERM, UMR-S U1297 and University of Toulouse III, Institute of Cardiovascular and Metabolic Diseases (I2MC), CHU-Rangueil, Toulouse, France
Toulouse, Languedoc-Roussillon, France
AL

Agustin Lahoz

Professor, PhD
Biomarkers and Precision Medicine Unit, Instituto de Investigación Sanitaria Fundación Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
Mk

Marcin kortylewski

Professor, PhD
Center for Gene Therapy and Department of Immuno-Oncology, Beckman Research Institute at City of Hope Comprehensive Cancer Center, Duarte, California, US
Duarte, California, United States
RG

Rocio Gonzalez-Conejero

Professor, PhD
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, Universidad de Murcia, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain
Constantino Martinez, PhD

Investigator
Servicio de Hematología y Oncología Médica, Hospital Universitario Morales Meseguer, Centro Regional de Hemodonación, IMIB-Pascual Parrilla, Murcia, Spain
Murcia, Murcia, Spain

Background: The 50% reduction of miR-146a levels in rs2431697-TT individuals has been associated with cardiovascular events. Our previous results have shown that platelet miR146a levels might modify platelet function and metabolism.

Aims: We now investigated the anti-thrombotic potential of miR-146a as therapeutic tool.

Methods: Platelet function from WT and miR-146a-/- mice were evaluated. Pulmonary embolism (PE) murine model was performed after miR-146a-mimic injection. T-TAS was used in human platelets.

Results: miR-146a-/- platelet hyperreactivity was observed with most agonists. miR-146a-/- platelets displayed more α-granules, enhanced F-actin formation by Thr and spreading on fibrinogen. P2y1 expression (miR-146a predicted target) and ADP activation was higher in miR-146-/- platelets. Interestingly, we previously observed that oxygen consumption rate (OCR) was greater in miR-146a-/- vs. WT displaying increased maximal and spare respiratory capacities and metabolic analysis showed elevated carnitines and decreased FA levels. We now showed that etomoxir, an in vitro Cpt1a inhibitor, abolished differences observed in the OCR and in the microfluids between WT and deficient platelets, while palmitate increased more the OCR in miR-146a-/- platelets. miR-146a-/- mice had higher mortality rate (43.3% vs. 19.2% in WT) in PE model and presented larger aggregates in lungs. Importantly, miR-146a-/- mice injected with miR-146a-mimic were fully protected against PE (100% survival), while RNA-scramble injection had no impact on mice survival (44.4%). All of these murine results were supported by those obtained in healthy rs2431697-TT subjects showing a significantly lower starting occlusion time and higher reactivity with TRAP-6 than CC individuals.

Conclusion(s): miR-146a deficiency causes a metabolic shift to FA β-oxidation provoking a hyperreactive phenotype in human and mouse platelets. Thus, miR-146-/- mice presented reduced survival rate after PE that was recovered with miR-146a-mimic therapy due to the rise of this miRNA in platelets. Therefore, miR-146a is a crucial regulator of platelet function and it might be a new antithrombotic therapeutic approach.

Learning Objectives:

Platelet function analysis: platelet activation by flow cytometry, spreading, transmission electron microscopy, aggregometry, microfluidics.
Platelet metabolism: studies using Seahorse Analyzer and untargeted metabolomics in platelets and serum.
Thrombosis model: pulmonary thromboembolism model in mice injecting epinephrine and collagen.