Stiffness-Induced Endothelial DLC-1 Expression Forces Leukocyte Spreading through Stabilization of the ICAM-1 Adhesome
Lilian Schimmel,
Miesje van der Stoel,
Carmela Rianna,
Anne-Marieke van Stalborch,
Aafke de Ligt,
Mark Hoogenboezem,
Simon Tol,
Jos van Rijssel,
Robert Szulcek,
Harm Jan Bogaard,
Patrick Hofmann,
Reinier Boon,
Manfred Radmacher,
Vivian de Waard,
Stephan Huveneers,
Jaap D. van Buul
Affiliations
Lilian Schimmel
Molecular Cell Biology Laboratory, Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Miesje van der Stoel
Department of Medical Biochemistry, Amsterdam Cardiovascular Sciences, Academic Medical Center, 1105 AZ Amsterdam, the Netherlands
Carmela Rianna
Biophysics Institute, University of Bremen, D-28334 Bremen, Germany
Anne-Marieke van Stalborch
Molecular Cell Biology Laboratory, Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Aafke de Ligt
Molecular Cell Biology Laboratory, Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Mark Hoogenboezem
Departmental Central Facility, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Simon Tol
Departmental Central Facility, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Jos van Rijssel
Molecular Cell Biology Laboratory, Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands
Robert Szulcek
Department of Pulmonary Diseases, VU Medical Center, 1081 HV Amsterdam, the Netherlands
Harm Jan Bogaard
Department of Pulmonary Diseases, VU Medical Center, 1081 HV Amsterdam, the Netherlands
Patrick Hofmann
Department of Physiology, VU Medical Center, Amsterdam, the Netherlands
Reinier Boon
Department of Physiology, VU Medical Center, Amsterdam, the Netherlands
Manfred Radmacher
Biophysics Institute, University of Bremen, D-28334 Bremen, Germany
Vivian de Waard
Department of Medical Biochemistry, Amsterdam Cardiovascular Sciences, Academic Medical Center, 1105 AZ Amsterdam, the Netherlands
Stephan Huveneers
Department of Medical Biochemistry, Amsterdam Cardiovascular Sciences, Academic Medical Center, 1105 AZ Amsterdam, the Netherlands
Jaap D. van Buul
Molecular Cell Biology Laboratory, Department of Plasma Proteins, Sanquin Research and Landsteiner Laboratory, Academic Medical Center, University of Amsterdam, 1066 CX Amsterdam, the Netherlands; Corresponding author
Summary: Leukocytes follow the well-defined steps of rolling, spreading, and crawling prior to diapedesis through endothelial cells (ECs). We found increased expression of DLC-1 in stiffness-associated diseases like atherosclerosis and pulmonary arterial hypertension. Depletion of DLC-1 in ECs cultured on stiff substrates drastically reduced cell stiffness and mimicked leukocyte transmigration kinetics observed for ECs cultured on soft substrates. Mechanistic studies revealed that DLC-1-depleted ECs or ECs cultured on soft substrates failed to recruit the actin-adaptor proteins filamin B, α-actinin-4, and cortactin to clustered ICAM-1, thereby preventing the ICAM-1 adhesome formation and impairing leukocyte spreading. This was rescued by overexpressing DLC-1, resulting in ICAM-1 adhesome stabilization and leukocyte spreading. Our results reveal an essential role for substrate stiffness-regulated endothelial DLC-1, independent of its GAP domain, in locally stabilizing the ICAM-1 adhesome to promote leukocyte spreading, essential for efficient leukocyte transendothelial migration. : Leukocyte extravasation depends on local cellular and substrate stiffness. Schimmel et al. identified endothelial DLC-1 as a mediator to translate stiffness to leukocyte behavior. DLC-1 is crucial for the ICAM-1 adhesome, which allows leukocytes to switch from the rolling to the spreading and crawling phase, followed by diapedesis. Keywords: ICAM-1, DLC-1, spreading, leukocyte, transmigration, diapedesis, rolling, stiffness, mechanosignaling, endothelial