GRP78 is a novel receptor initiating a vascular barrier protective response to oxidized phospholipids.
Vascular integrity and the maintenance of blood vessel continuity are fundamental features of the circulatory system maintained through endothelial cell-cell junctions. Defects in the endothelial barrier become an initiating factor in several pathologies, including ischemia/reperfusion, tumor angiogenesis, pulmonary edema, sepsis, and acute lung injury. Better understanding of mechanisms stimulating endothelial barrier enhancement may provide novel therapeutic strategies. We previously reported that oxidized phospholipids (oxidized 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine [OxPAPC]) promote endothelial cell (EC) barrier enhancement both in vitro and in vivo. This study examines the initiating mechanistic events triggered by OxPAPC to increase vascular integrity. Our data demonstrate that OxPAPC directly binds the cell membrane-localized chaperone protein, GRP78, associated with its cofactor, HTJ-1. OxPAPC binding to plasma membrane-localized GRP78 leads to GRP78 trafficking to caveolin-enriched microdomains (CEMs) on the cell surface and consequent activation of sphingosine 1-phosphate receptor 1, Src and Fyn tyrosine kinases, and Rac1 GTPase, processes essential for cytoskeletal reorganization and EC barrier enhancement. Using animal models of acute lung injury with vascular hyperpermeability, we observed that HTJ-1 knockdown blocked OxPAPC protection from interleukin-6 and ventilator-induced lung injury. Our data indicate for the first time an essential role of GRP78 and HTJ-1 in OxPAPC-mediated CEM dynamics and enhancement of vascular integrity.
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Anna A
AA
Birukova
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Patrick A
PA
Singleton
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Grzegorz
G
Gawlak
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Xinyong
X
Tian
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Tamara
T
Mirzapoiazova
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Bolot
B
Mambetsariev
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637.
Oleksii
O
Dubrovskyi
Lung Injury Center, Section of Pulmonary and Critical Care, Department of Medicine, Division of Biomedical Sciences, University of Chicago, Chicago, IL 60637Department of Vascular Biology and Thrombosis Research, Medical University of Vienna, 1090 Vienna, Austria.
Olga V
OV
Oskolkova
Department of Vascular Biology and Thrombosis Research, Medical University of Vienna, 1090 Vienna, Austria.
Valery N
VN
Bochkov
Department of Vascular Biology and Thrombosis Research, Medical University of Vienna, 1090 Vienna, Austria kbirukov@medicine.bsd.uchicago.edu.
Konstantin G
KG
Birukov
D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D022461:Caveolins / Q000378:metabolism; D002478:Cells, Cultured; D017097:Electric Impedance; D000091342:Endoplasmic Reticulum Chaperone BiP; D042783:Endothelial Cells / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology; D050956:HSP40 Heat-Shock Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000502:physiology*; D006801:Humans; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008810:Mice, Inbred C57BL; D010084:Oxidation-Reduction; D010713:Phosphatidylcholines / Q000502:physiology*; D021381:Protein Transport; D011651:Pulmonary Artery / Q000166:cytology; D049349:Receptors, Lysosphingolipid / Q000378:metabolism
D000199:Actins; D022461:Caveolins; C485368:DNAJC1 protein, human; D000091342:Endoplasmic Reticulum Chaperone BiP; D050956:HSP40 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D008565:Membrane Proteins; D010713:Phosphatidylcholines; D049349:Receptors, Lysosphingolipid; C472349:oxidized-L-alpha-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine