https://orcid.org/0000-0001-7156-5365
FVIIa expressed by myeloid cells is crucial for integrin αMβ2–dependent migration on fibrin.
TF-FVIIa induces PAR2–β-arrestin–biased signaling in monocytes and macrophages.
Abstract
Protease activated receptors (PARs) are cleaved by coagulation proteases and thereby connect hemostasis with innate immune responses. Signaling of the tissue factor (TF) complex with factor VIIa (FVIIa) via PAR2 stimulates extracellular signal-regulated kinase (ERK) activation and cancer cell migration, but functions of cell autonomous TF-FVIIa signaling in immune cells are unknown. Here, we show that myeloid cell expression of FVII but not of FX is crucial for inflammatory cell recruitment to the alveolar space after challenge with the double-stranded viral RNA mimic polyinosinic:polycytidylic acid [Poly(I:C)]. In line with these data, genetically modified mice completely resistant to PAR2 cleavage but not FXa-resistant PAR2–mutant mice are protected from lung inflammation. Poly(I:C)-stimulated migration of monocytes/macrophages is dependent on ERK activation and mitochondrial antiviral signaling (MAVS) but independent of toll-like receptor 3 (TLR3). Monocyte/macrophage-synthesized FVIIa cleaving PAR2 is required for integrin αMβ2-dependent migration on fibrinogen but not for integrin β1-dependent migration on fibronectin. To further dissect the downstream signaling pathway, we generated PAR2S365/T368A-mutant mice deficient in β-arrestin recruitment and ERK scaffolding. This mutation reduces cytosolic, but not nuclear ERK phosphorylation by Poly(I:C) stimulation, and prevents macrophage migration on fibrinogen but not fibronectin after stimulation with Poly(I:C) or CpG-B, a single-stranded DNA TLR9 agonist. In addition, PAR2S365/T368A-mutant mice display markedly reduced immune cell recruitment to the alveolar space after Poly(I:C) challenge. These results identify TF-FVIIa-PAR2-β-arrestin–biased signaling as a driver for lung infiltration in response to viral nucleic acids and suggest potential therapeutic interventions specifically targeting TF-VIIa signaling in thrombo-inflammation.
References
1.
Taylor
FB
, Chang
A
, Ruf
W
, et al. Lethal E. coli septic shock is prevented by blocking tissue factor with monoclonal antibody
. Circ Shock
. 1991
;33
(3
):127
-134
.2.
Antoniak
S
, Mackman
N
. Multiple roles of the coagulation protease cascade during virus infection
. Blood
. 2014
;123
(17
):2605
-2613
.3.
Pawlinski
R
, Wang
JG
, Owens
AP
, et al. Hematopoietic and nonhematopoietic cell tissue factor activates the coagulation cascade in endotoxemic mice
. Blood
. 2010
;116
(5
):806
-814
.4.
Ebert
J
, Wilgenbus
P
, Teiber
JF
, et al. Paraoxonase-2 regulates coagulation activation through endothelial tissue factor
. Blood
. 2018
;131
(19
):2161
-2172
.5.
Skendros
P
, Mitsios
A
, Chrysanthopoulou
A
, et al. Complement and tissue factor-enriched neutrophil extracellular traps are key drivers in COVID-19 immunothrombosis
. J Clin Invest
. 2020
;130
(11
):6151
-6157
.6.
Shaver
CM
, Grove
BS
, Putz
ND
, et al. Regulation of alveolar procoagulant activity and permeability in direct acute lung injury by lung epithelial tissue factor
. Am J Respir Cell Mol Biol
. 2015
;53
(5
):719
-727
.7.
Kral-Pointner
JB
, Schrottmaier
WC
, Horvath
V
, et al. Myeloid but not epithelial tissue factor exerts protective anti-inflammatory effects in acid aspiration-induced acute lung injury
. J Thromb Haemost
. 2017
;15
(8
):1625
-1639
.8.
Zelaya
H
, Rothmeier
AS
, Ruf
W
. Tissue factor at the crossroad of coagulation and cell signaling
. J Thromb Haemost
. 2018
;16
(10
):1941
-1952
.9.
Moretti
S
, Bellocchio
S
, Bonifazi
P
, et al. The contribution of PARs to inflammation and immunity to fungi
. Mucosal Immunol
. 2008
;1
(2
):156
-168
.10.
Antoniak
S
, Tatsumi
K
, Schmedes
CM
, et al. PAR1 regulation of CXCL1 expression and neutrophil recruitment to the lung in mice infected with influenza A virus
. J Thromb Haemost
. 2021
;19
(4
):1103
-1111
.11.
Gunther
RC
, Bharathi
V
, Miles
SD
, et al. Myeloid protease-activated receptor-2 contributes to influenza a virus pathology in mice
. Front Immunol
. 2021
;12
:791017
.12.
Rayees
S
, Joshi
JC
, Tauseef
M
, et al. PAR2-mediated cAMP generation suppresses TRPV4-dependent Ca(2+) signaling in alveolar macrophages to resolve TLR4-induced inflammation
. Cell Rep
. 2019
;27
(3
):793
-805.e4
.13.
Nhu
QM
, Shirey
KA
, Pennini
ME
, Stiltz
J
, Vogel
SN
. Proteinase-activated receptor 2 activation promotes an anti-inflammatory and alternatively activated phenotype in LPS-stimulated murine macrophages
. Innate Immun
. 2012
;18
(2
):193
-203
.14.
Disse
J
, Ruf
W
. Endothelial protein C receptor is required for tissue factor ternary complex signaling in the mouse
. J Thromb Haemost
. 2011
;9
(12
):2516
-2518
.15.
Liang
HP
, Kerschen
EJ
, Hernandez
I
, et al. EPCR-dependent PAR2 activation by the blood coagulation initiation complex regulates LPS-triggered interferon responses in mice
. Blood
. 2015
;125
(18
):2845
-2854
.16.
Muller-Calleja
N
, Hollerbach
A
, Royce
J
, et al. Lipid presentation by the protein C receptor links coagulation with autoimmunity
. Science
. 2021
;371
(6534
):eabc0956
.17.
Nhu
QM
, Shirey
K
, Teijaro
JR
, et al. Novel signaling interactions between proteinase-activated receptor 2 and toll-like receptors in vitro and in vivo
. Mucosal Immunol
. 2010
;3
(1
):29
-39
.18.
Graf
C
, Wilgenbus
P
, Pagel
S
, et al. Myeloid cell-synthesized coagulation factor X dampens antitumor immunity
. Sci Immunol
. 2019
;4
(39
):eaaw8405
.19.
Fleischer
MI
, Röhrig
N
, Raker
VK
, et al. Protease- and cell type-specific activation of protease-activated receptor 2 in cutaneous inflammation
. J Thromb Haemost
. 2022
;20
(12
):2823
-2836
.20.
Rothmeier
AS
, Liu
E
, Chakrabarty
S
, et al. Identification of the integrin-binding site on coagulation factor VIIa required for proangiogenic PAR2 signaling
. Blood
. 2018
;131
(6
):674
-685
.21.
DeFea
KA
, Zalevsky
J
, Thoma
MS
, Déry
O
, Mullins
RD
, Bunnett
N
. B-arrestin-dependent endocytosis of proteinase-activated receptor 2 is required for intracellular targeting of activated ERK1/2
. J Cell Biol
. 2000
;148
(6
):1267
-1281
.22.
Ge
L
, Ly
Y
, Hollenberg
M
, DeFea
K
. A b-arrestin-dependent scaffold is associated with prolonged MAPK activation in pseudopodia during protease-activated receptor-2 induced chemotaxis
. J Biol Chem
. 2003
;278
(36
):34418
-34426
.23.
Ge
L
, Shenoy
SK
, Lefkowitz
RJ
, DeFea
K
. Constitutive protease-activated receptor-2-mediated migration of MDA MB-231 breast cancer cells requires both beta-arrestin-1 and -2
. J Biol Chem
. 2004
;279
(53
):55419
-55424
.24.
Griffin
JH
, Zlokovic
BV
, Mosnier
LO
. Activated protein C: biased for translation
. Blood
. 2015
;125
(19
):2898
-2907
.25.
Sharma
A
, Kontodimas
K
, Bosmann
M
. The MAVS immune recognition pathway in viral infection and sepsis
. Antioxid Redox Signal
. 2021
;35
(16
):1376
-1392
.26.
Chapman
HA
, Allen
CL
, Stone
OL
, Fair
DS
. Human alveolar macrophages synthesize factor VII in vitro. Possible role in interstitial lung disease
. J Clin Invest
. 1985
;75
(6
):2030
-2037
.27.
Scotton
CJ
, Krupiczojc
MA
, Königshoff
M
, et al. Increased local expression of coagulation factor X contributes to the fibrotic response in human and murine lung injury
. J Clin Invest
. 2009
;119
(9
):2550
-2563
.28.
Zhang
N
, Czepielewski
RS
, Jarjour
NN
, et al. Expression of factor V by resident macrophages boosts host defense in the peritoneal cavity
. J Exp Med
. 2019
;216
(6
):1291
-1300
.29.
Schaffner
F
, Yokota
N
, Carneiro-Lobo
TC
, et al. Endothelial protein c receptor function in murine and human breast cancer development
. PLoS One
. 2013
;8
(4
):e61071
.30.
Müller-Calleja
N
, Hollerbach
A
, Ritter
S
, et al. Tissue factor pathway inhibitor primes monocytes for antiphospholipid antibody-induced thrombosis
. Blood
. 2019
;134
(14
):1119
-1131
.31.
Kamikubo
Y
, Mendolicchio
GL
, Zampolli
A
, et al. Selective factor VIII activation by the tissue factor-factor VIIa-factor Xa complex
. Blood
. 2017
;130
(14
):1661
-1670
.32.
Zelaya
H
, Tada
A
, Vizoso-Pinto
MG
, et al. Nasal priming with immunobiotic Lactobacillus rhamnosus modulates inflammation-coagulation interactions and reduces influenza virus-associated pulmonary damage
. Inflamm Res
. 2015
;64
(8
):589
-602
.33.
Cao
C
, Lawrence
DA
, Li
Y
, et al. Endocytic receptor LRP together with tPA and PAI-1 coordinates Mac-1-dependent macrophage migration
. EMBO J
. 2006
;25
(9
):1860
-1870
.34.
Gautier
EL
, Shay
T
, Miller
J
, et al. Gene-expression profiles and transcriptional regulatory pathways that underlie the identity and diversity of mouse tissue macrophages
. Nat Immunol
. 2012
;13
(11
):1118
-1128
.35.
Antonelli
LR
, Gigliotti Rothfuchs
A
, Gonçalves
R
, et al. Intranasal Poly-IC treatment exacerbates tuberculosis in mice through the pulmonary recruitment of a pathogen-permissive monocyte/macrophage population
. J Clin Invest
. 2010
;120
(5
):1674
-1682
.36.
Yu
YR
, O'Koren
EG
, Hotten
DF
, et al. A protocol for the comprehensive flow cytometric analysis of immune cells in normal and inflamed murine non-lymphoid tissues
. PLoS One
. 2016
;11
(3
):e0150606
.37.
Garré
JM
, Silva
HM
, Lafaille
JJ
, Yang
G
. CX3CR1(+) monocytes modulate learning and learning-dependent dendritic spine remodeling via TNF-alpha
. Nat Med
. 2017
;23
(6
):714
-722
.38.
Saffarzadeh
M
, Grunz
K
, Nguyen
TS
, et al. Macrophage protease-activated receptor 2 regulates fetal liver erythropoiesis in mice
. Blood Adv
. 2020
;4
(22
):5810
-5824
.39.
Dorfleutner
A
, Hintermann
E
, Tarui
T
, Takada
Y
, Ruf
W
. Crosstalk of integrin a3b1 and tissue factor in cell migration
. Mol Biol Cell
. 2004
;15
(10
):4416
-4425
.40.
Flick
MJ
, Du
X
, Witte
DP
, et al. Leukocyte engagement of fibrin(ogen) via the integrin receptor alphaMbeta2/Mac-1 is critical for host inflammatory response in vivo
. J Clin Invest
. 2004
;113
(11
):1596
-1606
.41.
Flick
MJ
, LaJeunesse
CM
, Talmage
KE
, et al. Fibrin(ogen) exacerbates inflammatory joint disease through a mechanism linked to the integrin alphaMbeta2 binding motif
. J Clin Invest
. 2007
;117
(11
):3224
-3235
.42.
Rothmeier
AS
, Marchese
P
, Langer
F
, et al. Tissue factor prothrombotic activity is regulated by integrin-arf6 trafficking
. Arterioscler Thromb Vasc Biol
. 2017
;37
(7
):1323
-1331
.43.
Ahn
GO
, Tseng
D
, Liao
CH
, Dorie
MJ
, Czechowicz
A
, Brown
JM
. Inhibition of Mac-1 (CD11b/CD18) enhances tumor response to radiation by reducing myeloid cell recruitment
. Proc Natl Acad Sci U S A
. 2010
;107
(18
):8363
-8368
.44.
Cheng
RKY
, Fiez-Vandal
C
, Schlenker
O
, et al. Structural insight into allosteric modulation of protease-activated receptor 2
. Nature
. 2017
;545
(7652
):112
-115
.45.
Stassen
JM
, Lambeir
A-M
, Matthyssens
G
, et al. Characterisation of a novel series of aprotinin-derived anticoagulants. I. In vitro and pharmacological properties
. Thromb Haemost
. 1995
;74
(2
):646
-654
.46.
Seth
RB
, Sun
L
, Ea
CK
, Chen
ZJ
. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3
. Cell
. 2005
;122
(5
):669
-682
.47.
Ott
I
, Weigand
B
, Michl
R
, et al. Tissue factor cytoplasmic domain stimulates migration by activation of the GTPase Rac1 and the mitogen-activated protein kinase p38
. Circulation
. 2005
;111
(3
):349
-355
.48.
Garlapati
V
, Molitor
M
, Michna
T
, et al. Targeting myeloid cell coagulation signaling blocks MAP kinase/TGF-beta1-driven fibrotic remodeling in ischemic heart failure
. J Clin Invest
. 2023
;133
(4
):e156436
.49.
Schaffner
F
, Versteeg
HH
, Schillert
A
, et al. Cooperation of tissue factor cytoplasmic domain and PAR2 signaling in breast cancer development
. Blood
. 2010
;116
(26
):6106
-6113
.50.
Nichols
HL
, Saffeddine
M
, Theriot
BS
, et al. Beta-arrestin-2 mediates the proinflammatory effects of proteinase-activated receptor-2 in the airway
. Proc Natl Acad Sci U S A
. 2012
;109
(41
):16660
-16665
.51.
Koizume
S
, Jin
M-S
, Miyagi
E
, et al. Activation of cancer cell migration and invasion by ectopic synthesis of coagulation factor VII
. Cancer Res
. 2006
;66
(19
):9453
-9460
.52.
Ünlü
B
, Kocatürk
B
, Rondon
AMR
, et al. Integrin regulation by tissue factor promotes cancer stemness and metastatic dissemination in breast cancer
. Oncogene
. 2022
;41
(48
):5176
-5185
.53.
Tsai
MC
, Chen
KD
, Wang
CC
, et al. Factor VII promotes hepatocellular carcinoma progression through ERK-TSC signaling
. Cell Death Discov
. 2015
;1
:15051
.54.
Subramaniam
S
, Ogoti
Y
, Hernandez
I
, et al. A thrombin-PAR1/2 feedback loop amplifies thromboinflammatory endothelial responses to the viral RNA analogue poly(I:C)
. Blood Adv
. 2021
;5
(13
):2760
-2774
.55.
Antoniak
S
, Owens
AP
, Baunacke
M
, et al. PAR-1 contributes to the innate immune response during viral infection
. J Clin Invest
. 2013
;123
(3
):1310
-1322
.56.
Lavoie
H
, Gagnon
J
, Therrien
M
. ERK signalling: a master regulator of cell behaviour, life and fate
. Nat Rev Mol Cell Biol
. 2020
;21
(10
):607
-632
.57.
Celi
A
, Pellegrini
G
, Lorenzet
R
, et al. P-selectin induces the expression of tissue factor on monocytes
. Proc Natl Acad Sci U S A
. 1994
;91
(19
):8767
-8771
.58.
Randolph
GJ
, Luther
T
, Albrecht
S
, Magdolen
V
, Muller
WA
. Role of tissue factor in adhesion of mononuclear phagocytes to and trafficking through endothelium in vitro
. Blood
. 1998
;92
(11
):4167
-4177
.59.
Geiger
H
, Pawar
SA
, Kerschen
EJ
, et al. Pharmacological targeting of the thrombomodulin-activated protein C pathway mitigates radiation toxicity
. Nat Med
. 2012
;18
(7
):1123
-1129
.60.
Nguyen
TS
, Lapidot
T
, Ruf
W
. Extravascular coagulation in hematopoietic stem and progenitor cell regulation
. Blood
. 2018
;132
(2
):123
-131
.© 2024 American Society of Hematology. Published by Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.
2024
You do not currently have access to this content.
This feature is available to Subscribers Only
Sign In or Create an Account Close Modal