PB0396 - Characterization of thrombogenic effects of platelet tyrosine phosphatase SHP2 inhibition using the allosteric inhibitor SHP099

Sunday, July 10, 2022

6:30 PM – 7:30 PM

Location: Exhibition

Presentation Format: In-Person

Presenting Author(s)

Delia I. Fernández de la Fuente, MSc

PhD Student
School for Cardiovascular Diseases (CARIM), Maastricht University, 6200 MD Maastricht, The Netherlands
Maastricht, Limburg, Netherlands

Co-Author(s)

Lidia Hermida-Nogueira

PhD
Platelet Proteomics Group, Center for Research in Molecular Medicine and Chronic Diseases (CiMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain
Santiago de Compostela, Galicia, Spain
JH

Jingnan Huang

PhD Student
Department of Biochemistry, CARIM, Maastricht University, 6200 MD Maastricht, The Netherlands
Maastricht, Limburg, Netherlands
SV

Sonia Veiras

Medical Doctor
Department of Anesthesiology and Intensive Care Medicine, Clinical University Hospital of Santiago, Santiago de Compostela, Spain
Santiago de Compostela, Galicia, Spain
MK

Marijke Kuijpers

Associate professor
Maastricht University
Maastricht, Limburg, Netherlands
JW

Johan W. M. Heemskerk

Emeritus Professor
Synapse research insitute
MAASTRICHT, Limburg, Netherlands
AG

Angel Garcia

Professor
Universidade de Santiago de Compostela
Santiago de Compostela, Galicia, Spain

Background: Protein tyrosine phosphatase SHP2 is a known regulator of platelet signal transduction potentially suppressing GPVI-induced activation. Currently, there are clinical trials with derivatives of the allosteric drug SHP099, which inhibits SHP2, as therapy for solid cancers. Human mutations of SHP2 gene, PTPN11, have been described to be linked to a variety of diseases.

Aims: Assessment of the potential thrombogenic effect of drug-induced SHP2 inhibition.

Methods: Washed human platelets were incubated with SHP099 and stimulated with the GPVI agonists collagen-related peptide (CRP) or collagen for aggregation and flow cytometric measurements. Drug effects of shear-dependent thrombus and fibrin formation were determined using whole-blood microfluidics over low and high collagen concentrations. In all assays, effects of co-inhibition of P2Y12 ADP receptor blocker were examined. Thromboelastography was used to evaluate the effect on clot formation.

Results: Pharmacological inhibition of SHP2 in washed platelets did not alter GPVI-dependent platelet aggregation regardless of the GPVI agonist used and P2Y12 inhibition. However, inhibition of SHP2 triggered faster integrin GPIIbIIIa opening, when assesed after GPVI activation. Moreover, in whole blood microfluidic assays, the drug enhanced the buildup of thrombus volume on collagen surfaces. When limiting the thrombus buildup by P2Y12 inhibition, this enhancement was abolished. In the presence of tissue factor and coagulation, the drug significantly enhanced thrombus volume, size, and contraction as well as fibrin formation. In thromboelastography, tissue factor-induced blood clotting profiles were enhanced with tranexamic acid, preventing fibrinolysis.

Conclusion(s): The thrombogenic effect of SHP2 inhibition is not confined to early GPVI activation and requires the presence of shear and/or coagulation.