LRRC8 complexes are ATP release channels that regulate platelet activation and arterial thrombosis
Abstract
Abstract Platelet shape and volume changes are early mechanical events contributing to platelet activation and thrombosis. Here, we identify single-nucleotide polymorphisms in leucine-rich repeat–containing 8 (LRRC8) protein subunits that form the volume-regulated anion channel (VRAC), which are independently associated with altered mean platelet volume. LRRC8A is required for functional VRAC in megakaryocytes (MKs) and regulates platelet volume; adhesion; and agonist-stimulated activation, aggregation, adenosine triphosphate (ATP) secretion, and calcium mobilization. MK-specific LRRC8A conditional knockout mice have reduced laser injury–induced cremaster arteriolar thrombus formation and prolonged FeCl3 induced carotid arterial thrombosis without prolonged bleeding times. Mechanistically, platelet LRRC8A mediates swell-induced cytosolic ATP release to amplify agonist-stimulated calcium–phosphoinositide 3-kinase–protein kinase B signaling. Small-molecule LRRC8 channel inhibitors recapitulate defects observed in LRRC8A-null platelets in vitro and in vivo. These studies identify the mechanoresponsive LRRC8 channel complex as an ATP release channel in platelets, which positively regulates platelet function and thrombosis, providing a proof of concept for a novel antithrombotic drug target.
Article Details
Authors (28)
John D. Tranter
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Ryan T. Mikami
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Ashutosh Kumar
Gavriel Brown
2Division of Hematology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
Tarek Mohamed Abd El-Aziz
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Yonghui Zhao
Center for Low-Carbon Conversion Science and Engineering; State Key Laboratory of Low Carbon Catalysis and Carbon Dioxide Utilization, Shanghai Advanced Research Institute
Prakash Arullampalam
Katrina Ashworth
1Washington University School of Medicine, Saint Louis, United States
Vishwanath Jha
2Division of Hematology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
Nihil Abraham
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Chloe Meyer
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Abigail Ajanel
3Department of Emergency Medicine, Washington University School of Medicine, St. Louis, MO
Litao Xie
Yongmei Feng
Juan Hong
Haixia Zhang
Tripti Kumari
2Division of Hematology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
Adam Balutowski
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Alice Liu
Division of Hematology, Department of Pediatrics, Washington University
David Bark
Division of Hematology, Department of Pediatrics, Washington University
Vinayak K. Nair
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO
Nina M. Lasky
4Division of Pediatric Hematology and Oncology, Washington University in St. Louis, St. Louis, MO
Nathan O. Stitziel
From the Cardiovascular Division, Department of Medicine, and the Department of Genetics, Washington University School of Medicine, St. Louis.
Daniel J. Lerner
8Senseion Therapeutics Inc, St. Louis, MO
Robert A. Campbell
Department of Emergency Medicine, School of Medicine, Washington University
Jorge Di Paola
Division of Hematology, Department of Pediatrics, Washington University
Jaehyung Cho
Rajan Sah
1Cardiovascular Division, Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO