TY - JOUR
T1 - Proteoglycan 4 Modulates Osteogenic Smooth Muscle Cell Differentiation during Vascular Remodeling and Intimal Calcification
AU - Seime, Till
AU - Akbulut, Asim Cengiz
AU - Liljeqvist, Moritz Lindquist
AU - Siika, Antti
AU - Jin, Hong
AU - Winski, Greg
AU - van Gorp, Rick H
AU - Karlöf, Eva
AU - Lengquist, Mariette
AU - Buckler, Andrew J
AU - Kronqvist, Malin
AU - Waring, Olivia J
AU - Lindeman, Jan H N
AU - Biessen, Erik A L
AU - Maegdefessel, Lars
AU - Razuvaev, Anton
AU - Schurgers, Leon J
AU - Hedin, Ulf
AU - Matic, Ljubica
N1 - Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/5/21
Y1 - 2021/5/21
N2 - Calcification is a prominent feature of late-stage atherosclerosis, but the mechanisms driving this process are unclear. Using a biobank of carotid endarterectomies, we recently showed that Proteoglycan 4 (PRG4) is a key molecular signature of calcified plaques, expressed in smooth muscle cell (SMC) rich regions. Here, we aimed to unravel the PRG4 role in vascular remodeling and intimal calcification. PRG4 expression in human carotid endarterectomies correlated with calcification assessed by preoperative computed tomographies. PRG4 localized to SMCs in early intimal thickening, while in advanced lesions it was found in the extracellular matrix, surrounding macro-calcifications. In experimental models, Prg4 was upregulated in SMCs from partially ligated ApoE-/- mice and rat carotid intimal hyperplasia, correlating with osteogenic markers and TGFb1. Furthermore, PRG4 was enriched in cells positive for chondrogenic marker SOX9 and around plaque calcifications in ApoE-/- mice on warfarin. In vitro, PRG4 was induced in SMCs by IFNg, TGFb1 and calcifying medium, while SMC markers were repressed under calcifying conditions. Silencing experiments showed that PRG4 expression was driven by transcription factors SMAD3 and SOX9. Functionally, the addition of recombinant human PRG4 increased ectopic SMC calcification, while arresting cell migration and proliferation. Mechanistically, it suppressed endogenous PRG4, SMAD3 and SOX9, and restored SMC markers' expression. PRG4 modulates SMC function and osteogenic phenotype during intimal remodeling and macro-calcification in response to TGFb1 signaling, SMAD3 and SOX9 activation. The effects of PRG4 on SMC phenotype and calcification suggest its role in atherosclerotic plaque stability, warranting further investigations.
AB - Calcification is a prominent feature of late-stage atherosclerosis, but the mechanisms driving this process are unclear. Using a biobank of carotid endarterectomies, we recently showed that Proteoglycan 4 (PRG4) is a key molecular signature of calcified plaques, expressed in smooth muscle cell (SMC) rich regions. Here, we aimed to unravel the PRG4 role in vascular remodeling and intimal calcification. PRG4 expression in human carotid endarterectomies correlated with calcification assessed by preoperative computed tomographies. PRG4 localized to SMCs in early intimal thickening, while in advanced lesions it was found in the extracellular matrix, surrounding macro-calcifications. In experimental models, Prg4 was upregulated in SMCs from partially ligated ApoE-/- mice and rat carotid intimal hyperplasia, correlating with osteogenic markers and TGFb1. Furthermore, PRG4 was enriched in cells positive for chondrogenic marker SOX9 and around plaque calcifications in ApoE-/- mice on warfarin. In vitro, PRG4 was induced in SMCs by IFNg, TGFb1 and calcifying medium, while SMC markers were repressed under calcifying conditions. Silencing experiments showed that PRG4 expression was driven by transcription factors SMAD3 and SOX9. Functionally, the addition of recombinant human PRG4 increased ectopic SMC calcification, while arresting cell migration and proliferation. Mechanistically, it suppressed endogenous PRG4, SMAD3 and SOX9, and restored SMC markers' expression. PRG4 modulates SMC function and osteogenic phenotype during intimal remodeling and macro-calcification in response to TGFb1 signaling, SMAD3 and SOX9 activation. The effects of PRG4 on SMC phenotype and calcification suggest its role in atherosclerotic plaque stability, warranting further investigations.
KW - ARTICULAR CHONDROCYTES
KW - ATHEROSCLEROTIC LESIONS
KW - BALLOON INJURY
KW - CAROTID PLAQUE
KW - COMPUTED-TOMOGRAPHY ANGIOGRAPHY
KW - GENE-EXPRESSION
KW - HISTOLOGICAL CLASSIFICATION
KW - NATURAL-HISTORY
KW - PHENOTYPIC MODULATION
KW - Proteoglycan 4
KW - ZONE PROTEIN SZP
KW - atherosclerosis
KW - calcification
KW - extracellular matrix
KW - smooth muscle cells
KW - vascular remodeling
U2 - 10.3390/cells10061276
DO - 10.3390/cells10061276
M3 - Article
C2 - 34063989
SN - 2073-4409
VL - 10
JO - Cells
JF - Cells
IS - 6
M1 - 1276
ER -