Integrin-αIIbβ3-mediated outside-in signalling activates a negative feedback pathway to suppress platelet activation

Baiyun Dai; Peng Wu; Feng Xue; Renchi Yang; Ziqiang Yu; Kesheng Dai; Changgeng Ruan; Gang Liu; Peter J. Newman; Cunji Gao

doi:10.1160/TH16-02-0096

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 116(05): 918-930
DOI: 10.1160/TH16-02-0096

Cellular Haemostasis and Platelets

Schattauer GmbH

Integrin-αIIbβ3-mediated outside-in signalling activates a negative feedback pathway to suppress platelet activation

Baiyun Dai

¹Chronic Disease Research Institute, Department of Nutrition and Food Hygiene, Zhejiang University School of Public Health, Hangzhou, China

,

Peng Wu

¹Chronic Disease Research Institute, Department of Nutrition and Food Hygiene, Zhejiang University School of Public Health, Hangzhou, China

,

Feng Xue

²State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC, Tianjin , China

,

Renchi Yang

²State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, CAMS & PUMC, Tianjin , China

,

Ziqiang Yu

³Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Jiangsu, China

,

Kesheng Dai

³Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Jiangsu, China

,

Changgeng Ruan

³Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Jiangsu, China

,

Gang Liu

¹Chronic Disease Research Institute, Department of Nutrition and Food Hygiene, Zhejiang University School of Public Health, Hangzhou, China

,

Peter J. Newman^*

⁴Medical College of Wisconsin; Milwaukee, Wisconsin, USA

⁵Blood Research Institute, Blood Center of Wisconsin; Milwaukee, Wisconsin, USA

,

Cunji Gao^*

¹Chronic Disease Research Institute, Department of Nutrition and Food Hygiene, Zhejiang University School of Public Health, Hangzhou, China

⁵Blood Research Institute, Blood Center of Wisconsin; Milwaukee, Wisconsin, USA

› Author Affiliations

Abstract

Summary

Integrin-αIIbβ3-mediated outside-in signalling is widely accepted as an amplifier of platelet activation; accumulating evidence suggests that outside-in signalling can, under certain conditions, also function as an inhibitor of platelet activation. The role of integrin-αIIbβ3-mediated outside-in signalling in platelet activation is disputable. We employed flow cytometry, aggregometry, immunoprecipitation, and immunoblotting to investigate the role of integrin-αIIbβ3-mediated outside-in signalling in platelet activation. Integrin αIIbβ3 inhibition enhances agonist-induced platelet ATP secretion. Human platelets lacking expression of αIIbβ3 exhibited more platelet ATP secretion than their wild-type counterparts. Moreover, integrin-αIIbβ3-mediated outside-in signals activate SHIP-1, which in turn mediates p-Akt dep-hosphorylation, leading to inactivation of PI3K/Akt signalling. Furthermore, 3AC (SHIP-1 inhibitor) inhibits platelet disaggregation, and promotes platelet ATP secretion. Upon ADP stimulation, Talin is recruited to αIIbβ3, and it is dissociated from αIIbβ3 when platelets disaggregate. In addition, treatment with RUC2, an inhibitor of αIIbβ3, which blocks αIIbβ3-mediated outside-in signalling, can markedly prevent the dissociation of talin from integrin. SHIP1 Inhibitor 3AC inhibits the dissociation of talin from integrin-β3. These results suggest that integrin-αIIbβ3-mediated outside-in signalling can serve as a brake to restrict unnecessary platelet activation by activated SHIP-1, which mediated the disassociation of talin from β3, leading to integrin inactivation and blocking of PI3K/Akt signalling to restrict platelet ATP secretion.

Keywords

Integrins - platelets - ATP - RGD-peptide - talin

Full Text

References

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