Discoidin Domain Receptor 2 Inhibits Fibrillogenesis of Collagen Type 1

Mihai, Cosmin; Iscru, Daniel F.; Druhan, Lawrence J.; Elton, Terry S.; Agarwal, Gunjan

doi:10.1016/j.jmb.2006.06.067

Cited by 43 publications

(61 citation statements)

References 40 publications

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“…15 We have demonstrated that the exogenous addition of recombinant DDR2 protein increased DDR2 density by fluorescence microscope (supplemental Figure I, available at http:/atvb.ahajournals.org). The increased DDR2 density may increase the collagen-DDR2 binding activity.…”

Section: Selection and Characterization Of Effective Anti-ddr2 Sirnasmentioning

confidence: 84%

RNA Interference for Discoidin Domain Receptor 2 Attenuates Neointimal Formation in Balloon Injured Rat Carotid Artery

Shyu¹,

Wang

Kuan

et al. 2008

ATVB

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Objectives-Discoidin domain receptor 2 (DDR2) plays potential roles in the regulation of collagen turnover mediated by smooth muscle cells (SMCs) in atherosclerosis. Little is known about the function of DDR2 in vascular system. We investigated whether inhibition of DDR2 by small interfering RNA (siRNA) can reduce neointimal formation after arterial injury. Key Words: discoidin domain receptor 2 Ⅲ smooth muscle cell Ⅲ RNA interference Ⅲ vascular injury D evelopment of neointimal thickening is the major cause of atherosclerosis and restensois. Smooth muscle cells (SMCs) are the predominant cell types in the media of blood vessel walls and contribute to neointimal formation after arterial injury. After arterial injury, SMCs synthesize collagens, 1,2 which act as important signaling molecule regulating SMC responses to arterial repair. 3 Discoidin domain receptor 1 (DDR1) and DDR2 have recently emerged as nonintegrintype receptors for collagen. 4 DDR1 is mainly expressed in epithelial cells, whereas DDR2 is found in mesenchymal cells. 5 DDR1 and DDR2 play potential roles in the regulation of collagen turnover mediated by vascular smooth muscle cells (VSMCs) in obstructive diseases of blood vessel. 6 Both DDRs were found to be highly expressed by SMCs within the fibrous cap. 6 DDR1 has been found to play an important role in mediating intimal thickening after arterial injury 3 ; however, little is known about the function of DDR2 in the vascular system. We have previously demonstrated that stretching of VSMCs in vitro resulted in upregulation of DDR2 expression. 7 In this study we hypothesize that DDR2 plays an important role in vascular injury in vivo. Methods and Results-SMCsRNA interference mediated by small interfering RNA (siRNA) can knock down gene expression at a translational level through interactions with its target messenger RNA 8 and shows great promise for therapeutic applications. 9 Here, we applied RNA interference to study the role of DDR2 in atherosclerosis in vitro and in vivo. Methods siRNA DesignedThe siRNA duplexes targeting the mouse DDR2 mRNA (GenBank accession no. NM_022563) were designed. For the initial screening of the most effective siRNA duplexes, the 21-nucleotide RNAs were synthesized by in vitro transcription (Dharmacon Inc). Further experiments were performed with the most efficient siRNA-duplex. The most efficient siRNA-duplex does have 1 mismatch to the rat sequence NM_031764. The mismatch 1 is located at the 6th base of the duplex, U for mouse and C for rat. No significant similarities to any genes other than DDR2 were found using BLAST against the rat reference sequence database. Vascular Smooth Muscle Cell CulturePrimary cultures of VSMCs were grown by the explant technique from the thoracic aorta of 220-to 260-g male Wistar rats, as Original

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Section: Selection and Characterization Of Effective Anti-ddr2 Sirnasmentioning

confidence: 84%

RNA Interference for Discoidin Domain Receptor 2 Attenuates Neointimal Formation in Balloon Injured Rat Carotid Artery

Shyu¹,

Wang

Kuan

et al. 2008

ATVB

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“…The stress/strain data obtained from each strip during the tensile ramp tests was fit to Fung's standard exponential model 32 :…”

Section: Mechanical Testingmentioning

confidence: 99%

“…27,32 The wet weight of the scleral strips was measured by first immersing the specimens in 0.9% saline for 5 minutes and then blotting dry the surfaces. Each specimen was measured three times using this protocol and the average was taken for further analysis.…”

Section: Biochemical Analysis Of Scleral Collagen Content and Solubilmentioning

confidence: 99%

“…27,32 Briefly, a small aliquot (10-30 lL) of each of the four sets of collagen (NSC, ASC, PSC, and ISC) extracted from each sclera specimen was hydrolyzed in a 4 N NaOH at 1218C for 30 minutes by autoclave. In addition to the tissue samples, four sets of hydroxyproline standards made from known increasing amounts of hydroxyproline solubilized in respective solvent also were hydrolyzed.…”

Section: Biochemical Analysis Of Scleral Collagen Content and Solubilmentioning

confidence: 99%

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Biomechanical Properties and Correlation With Collagen Solubility Profile in the Posterior Sclera of Canine Eyes With anADAMTS10Mutation

Palko

Iwabe

Pan

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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Citation: Palko JR, Iwabe S, Pan X, Agarwal G, Komáromy AM, Liu J. Biomechanical properties and correlation with collagen solubility profile in the posterior sclera of canine eyes with an ADAMTS10 mutation. Invest Ophthalmol Vis Sci. 2013;54:268554: -269554: . DOI:10.1167 PURPOSE. We examined the biomechanical properties and correlation with the collagen solubility profile of the posterior sclera in a canine model of primary open-angle glaucoma caused by the G661R missense mutation in the ADAMTS10 gene. METHODS.Scleral strips from ADAMTS10-mutant (affected) dogs and age-matched controls were collected. Viscoelastic properties (i.e., complex modulus and tan[d]) were measured using dynamic mechanical analysis (DMA) with a 0.15% sinusoidal strain at different frequencies superimposed upon different preloads. A tensile ramp was performed following DMA. The collagen solubility profile was examined using a colorimetric hydroxyproline assay to determine the amount of soluble and insoluble collagen. The viscoelastic properties were compared between groups using linear mixed models for repeated measures at different preloads and frequencies. The correlation between the biomechanical properties and collagen content were evaluated using Pearson correlations. RESULTS.Complex modulus and tan(d) were significantly lower in the affected group (P < 0.001), and the differences were consistent at different preloads and frequencies. The B value from the tensile ramp test also was significantly lower in the affected group (P ¼ 0.02). The insoluble collagen was significantly lower in the affected group (P < 0.05) and correlated positively with the complex modulus (R ¼ 0.88, P < 0.005).CONCLUSIONS. An inherently weaker and biochemically distinct posterior sclera was observed in dogs with the G661R missense mutation in ADAMTS10 before clinical indications of optic nerve damage. It remains to be shown whether and how the altered scleral biomechanics may affect the rate of glaucoma progression following intraocular pressure elevation.

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“…(33) Similar bone defects also were observed in a mouse colony with spontaneous autosomal-recessive mutation in the Ddr2 locus. (34) Of note, DDR2 was reported to exert effects on collagen fibrillogenesis in an osteoblast cell line, (35,36) suggesting the important role of DDR2 in this cell type. Thus our study sought to investigate DDR2 function and signaling in osteogenesis and chondrogenesis.…”

Section: Introductionmentioning

confidence: 99%

An essential role of discoidin domain receptor 2 (DDR2) in osteoblast differentiation and chondrocyte maturation via modulation of Runx2 activation

Zhang

et al. 2010

Journal of Bone and Mineral Research

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Discoidin domain receptor 2 (DDR2) belongs to receptor tyrosine kinase (RTK) family and is activated by collagen binding. Although the bone defects in Ddr2 null mice have been reported for a decade, the molecular mechanism remains unclear. This study sought to investigate the function and detailed mechanism of DDR2 in osteogenic and chondrogenic differentiation. Herein we found that in preosteoblastic cells, DDR2 activation was enhanced by osteogenic induction but was not paralleled with the alteration of DDR2 expression. Under differentiated condition, downregulation of endogenous DDR2 through specific shRNA dramatically repressed osteoblastic marker gene expression and osteogenic differentiation. Enforced expression of constitutively activated DDR2 increased the expression of bone markers in both undifferentiated and differentiated osteoblasts. Importantly, molecular evidence showed that DDR2 regulated the transactivity of Runx2, a master transcription factor involved in skeletal development, by modulating its phosphorylation. Analysis of candidate protein kinases indicated that extracellular signal-regulated kinase (ERK) activation is responsive to DDR2 signaling and involved in DDR2 regulation of Runx2 phosphorylation and transcriptional activity. Notably, a gain-of-function mutant of Runx2 with enhanced ERK-independent phosphorylation rescued the impaired osteogenic phenotypes observed in Ddr2-silenced cells, whereas a Runx2 mutant devoid of phosphorylation regulation by ERK inhibited DDR2 induction of osteogenesis. In addition, DDR2 facilitated Runx2 transactivation and type X collagen expression in hypertrophic chondrocytes. Thus this study reveals for the first time that DDR2 plays an essential role in osteoblast and chondrocyte differentiation. The mechanism disclosure may provide therapeutic targets for human genetic disorders caused by DDR2 deficiency. ß

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Discoidin Domain Receptor 2 Inhibits Fibrillogenesis of Collagen Type 1

Cited by 43 publications

References 40 publications

RNA Interference for Discoidin Domain Receptor 2 Attenuates Neointimal Formation in Balloon Injured Rat Carotid Artery

RNA Interference for Discoidin Domain Receptor 2 Attenuates Neointimal Formation in Balloon Injured Rat Carotid Artery

Biomechanical Properties and Correlation With Collagen Solubility Profile in the Posterior Sclera of Canine Eyes With anADAMTS10Mutation

An essential role of discoidin domain receptor 2 (DDR2) in osteoblast differentiation and chondrocyte maturation via modulation of Runx2 activation

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