BACKGROUND/AIMS: The endoplasmic reticulum (ER) stress protein, calreticulin (CRT), is required for the production of TGF-β-stimulated extracellular matrix (ECM) by fibroblasts. Since TGF-β regulates vascular fibroproliferative responses and collagen deposition, we investigated the effects of CRT knockdown on vascular smooth-muscle cell (VSMC) fibroproliferative responses and collagen deposition. METHODS: Using a carotid artery ligation model of vascular injury, Cre-recombinase-IRES-GFP plasmid was delivered with microbubbles (MB) to CRT-floxed mice using ultrasound (US) to specifically reduce CRT expression in the carotid artery. RESULTS: In vitro, Cre-recombinase-mediated CRT knockdown in isolated, floxed VSMCs decreased the CRT transcript and protein, and attenuated the induction of collagen I protein in response to TGF-β. TGF-β stimulation of collagen I was partly blocked by the NFAT inhibitor 11R-VIVIT. Following carotid artery ligation, CRT staining was upregulated with enhanced expression in the neointima 14-21 days after injury. Furthermore, Cre-recombinase-IRES-GFP plasmid delivered by targeted US reduced CRT expression in the neointima of CRT-floxed mice and led to a significant reduction in neointima formation and collagen deposition. The neointimal cell number was also reduced in mice, with a local, tissue-specific knockdown of CRT. CONCLUSIONS: This work establishes a novel role for CRT in mediating VSMC responses to injury through the regulation of collagen deposition and neointima formation.
BACKGROUND/AIMS: The endoplasmic reticulum (ER) stress protein, calreticulin (CRT), is required for the production of TGF-β-stimulated extracellular matrix (ECM) by fibroblasts. Since TGF-β regulates vascular fibroproliferative responses and collagen deposition, we investigated the effects of CRT knockdown on vascular smooth-muscle cell (VSMC) fibroproliferative responses and collagen deposition. METHODS: Using a carotid artery ligation model of vascular injury, Cre-recombinase-IRES-GFP plasmid was delivered with microbubbles (MB) to CRT-floxed mice using ultrasound (US) to specifically reduce CRT expression in the carotid artery. RESULTS: In vitro, Cre-recombinase-mediated CRT knockdown in isolated, floxed VSMCs decreased the CRT transcript and protein, and attenuated the induction of collagen I protein in response to TGF-β. TGF-β stimulation of collagen I was partly blocked by the NFAT inhibitor 11R-VIVIT. Following carotid artery ligation, CRT staining was upregulated with enhanced expression in the neointima 14-21 days after injury. Furthermore, Cre-recombinase-IRES-GFP plasmid delivered by targeted US reduced CRT expression in the neointima of CRT-floxed mice and led to a significant reduction in neointima formation and collagen deposition. The neointimal cell number was also reduced in mice, with a local, tissue-specific knockdown of CRT. CONCLUSIONS: This work establishes a novel role for CRT in mediating VSMC responses to injury through the regulation of collagen deposition and neointima formation.
Authors: T Kanzaki; K Tamura; K Takahashi; Y Saito; B Akikusa; H Oohashi; N Kasayuki; M Ueda; N Morisaki Journal: Arterioscler Thromb Vasc Biol Date: 1995-11 Impact factor: 8.311
Authors: Shirling Tsai; Scott T Hollenbeck; Evan J Ryer; Rachel Edlin; Dai Yamanouchi; Rishi Kundi; Chunjie Wang; Bo Liu; K Craig Kent Journal: Am J Physiol Heart Circ Physiol Date: 2009-06-12 Impact factor: 4.733
Authors: Lauren A Biwer; Miranda E Good; Kwangseok Hong; Rahul K Patel; Neha Agrawal; Robin Looft-Wilson; Swapnil K Sonkusare; Brant E Isakson Journal: Arterioscler Thromb Vasc Biol Date: 2017-11-09 Impact factor: 8.311
Authors: Claire Molony; Damien King; Mariana Di Luca; Michael Kitching; Abidemi Olayinka; Roya Hakimjavadi; Lourdes A N Julius; Emma Fitzpatrick; Yusof Gusti; Denise Burtenshaw; Killian Healy; Emma K Finlay; David Kernan; Andreu Llobera; Weimin Liu; David Morrow; Eileen M Redmond; Jens Ducrée; Paul A Cahill Journal: Stem Cell Rev Rep Date: 2021-03-17 Impact factor: 5.739