The Experts below are selected from a list of 5454 Experts worldwide ranked by ideXlab platform
Thomas N. Wight - One of the best experts on this subject based on the ideXlab platform.
-
The biochemistry and immunohistochemistry of Versican.
Methods in cell biology, 2017Co-Authors: Stephen P. Evanko, Charles W. Frevert, Christina K. Chan, Pamela Y. Johnson, Thomas N. WightAbstract:Versican is a chondroitin sulfate proteoglycan found in the extracellular matrix that is important for changes in cell phenotype associated with development and disease. Versican has been shown to be involved in cardiovascular disorders, as well as lung disease and fibrosis, inflammatory bowel disease, cancer, and several other diseases that have an inflammatory component. Versican was first identified as a fibroblast proteoglycan and forms large multimolecular complexes with hyaluronan and other components of the provisional matrix during wound healing and inflammation. The biology of Versican has been well studied. Versican plays a major role in embryogenesis, particularly heart formation, where Versican deletion proves lethal. The ability to purify Versican to characterize and to use in experimental systems is vital to defining its role in development and disease. Protein expression systems have proven challenging to obtain milligram quantities of full-length Versican. Here, we describe proteoglycan biochemical purification techniques that have been developed by others, but which we have adapted to use with our source tissues and cells. We also include methods for immunohistochemical localization and quantitation of Versican in tissue sections.
-
Versican: Role in Cancer Tumorigenesis
Extracellular Matrix in Tumor Biology, 2017Co-Authors: Paul A. Keire, Inkyung Kang, Thomas N. WightAbstract:Versican is an extracellular matrix proteoglycan that is expressed in a wide variety of cancers. Several cellular sources for Versican have been identified in a multitude of cancers including tumor cells, stromal cells, myeloid cells, and lymphoid cells. Versican plays a role in five of the six hallmarks of cancer including proliferative signaling, evasion of growth suppressor signaling, promotion of tissue invasion and metastasis, angiogenesis, and resistance to cell death. Versican also interacts with growth factors and cytokines to modify their activity and involvement in the cancer response. The synthesis and accumulation of Versican is regulated by similar pathways that regulate cancer progression, such as the canonical Wnt/β-catenin pathway and receptor tyrosine kinases. The expression and accumulation of Versican are associated with poor prognosis, disease progression, metastasis, and chemoresistance. A detailed analysis of the role of Versican in the disease course of leiomyosarcoma is provided here as an example of the importance of this extracellular matrix component in cancer pathogenesis. Collectively, our results and those from other groups suggest that Versican could serve as a point of control in the management and treatment of many cancers.
-
subepithelial accumulation of Versican in a cockroach antigen induced murine model of allergic asthma
Journal of Histochemistry and Cytochemistry, 2016Co-Authors: Stephen R Reeves, Charles W. Frevert, Mervyn J Merrilees, Gernot Kaber, Alyssa Sheih, Georgiana Cheng, Mark A Aronica, Jason S Debley, Steven F Ziegler, Thomas N. WightAbstract:The extracellular matrix (ECM) is an important contributor to the asthmatic phenotype. Recent studies investigating airway inflammation have demonstrated an association between hyaluronan (HA) accumulation and inflammatory cell infiltration of the airways. The ECM proteoglycan Versican interacts with HA and is important in the recruitment and activation of leukocytes during inflammation. We investigated the role of Versican in the pathogenesis of asthmatic airway inflammation. Using cockroach antigen (CRA)-sensitized murine models of allergic asthma, we demonstrate increased subepithelial Versican in the airways of CRA-treated mice that parallels subepithelial increases in HA and leukocyte infiltration. During the acute phase, CRA-treated mice displayed increased gene expression of the four major Versican isoforms, as well as increased expression of HA synthases. Furthermore, in a murine model that examines both acute and chronic CRA exposure, Versican staining peaked 8 days following CRA challenge and preceded subepithelial leukocyte infiltration. We also assessed Versican and HA expression in differentiated primary human airway epithelial cells from asthmatic and healthy children. Increases in the expression of Versican isoforms and HA synthases in these epithelial cells were similar to those of the murine model. These data indicate an important role for Versican in the establishment of airway inflammation in asthma.
-
inhibition of Versican expression by sirna facilitates tropoelastin synthesis and elastic fiber formation by human sk lms 1 leiomyosarcoma smooth muscle cells in vitro and in vivo
Matrix Biology, 2016Co-Authors: Paul Keire, Thomas N. Wight, Steven L Bressler, Inkyung Kang, Eileen R Mulvihill, Barry StarcherAbstract:Versican is an extracellular matrix (ECM) molecule that interacts with other ECM components to influence ECM organization, stability, composition, and cell behavior. Versican is known to increase in a number of cancers, but little is known about how Versican influences the amount and organization of the ECM components in the tumor microenvironment. In the present study, we modulated Versican expression using siRNAs in the human leiomyosarcoma (LMS) smooth muscle cell line SK-LMS-1, and observed the formation of elastin and elastic fibers in vitro and also in vivo in a nude mouse tumor model. Constitutive siRNA-directed knockdown of Versican in LMS cells resulted in increased levels of elastin, as shown by immunohistochemical staining of the cells in vitro, and by mRNA and protein analyses. Moreover, Versican siRNA LMS cells, when injected into nude mice, generated smaller tumors that had significantly greater immunohistochemical and histochemical staining for elastin when compared to control tumors. Additionally, microarray analyses were used to determine the influence of Versican isoform modulation on gene expression profiles, and to identify genes that influence and relate to the process of elastogenesis. cDNA microarray analysis and TaqMan low density array validation identified previously unreported genes associated with downregulation of Versican and increased elastogenesis. These results highlight an important role for the proteoglycan Versican in regulating the expression and assembly of elastin and the phenotype of LMS cells.
-
a role for Versican in the development of leiomyosarcoma
Journal of Biological Chemistry, 2014Co-Authors: Paul Keire, Steven L Bressler, Joan M Lemire, Badreddin Edris, Brian P Rubin, Maziar Rahmani, Bruce M Mcmanus, Matt Van De Rijn, Thomas N. WightAbstract:Leiomyosarcoma (LMS) is a mesenchymal cancer that occurs throughout the body. Although LMS is easily recognized histopathologically, the cause of the disease remains unknown. Versican, an extracellular matrix proteoglycan, increases in LMS. Microarray analyses of 80 LMSs and 24 leiomyomas showed a significant elevated expression of Versican in human LMS versus benign leiomyomas. To explore the importance of Versican in this smooth muscle cell tumor, we used Versican-directed siRNA to knock down Versican expression in a LMS human cell line, SK-LMS-1. Decreased Versican expression was accompanied by slower rates of LMS cell proliferation and migration, increased adhesion, and decreased accumulation of the extracellular matrix macromolecule hyaluronan. Addition of purified Versican to cells expressing Versican siRNA restored cell proliferation to the level of LMS controls, increased the pericellular coat and the retention of hyaluronan, and decreased cell adhesion in a dose-dependent manner. The presence of Versican was not only synergistic with hyaluronan in increasing cell proliferation, but the depletion of Versican decreased hyaluronan synthase expression and decreased the retention of hyaluronan. When LMS cells stably expressing Versican siRNA were injected into nude mice, the resulting tumors displayed significantly less Versican and hyaluronan staining, had lower volumes, and had reduced levels of mitosis as compared with controls. Collectively, these results suggest a role for using Versican as a point of control in the management and treatment of LMS.
Suneel S. Apte - One of the best experts on this subject based on the ideXlab platform.
-
Stromal Versican Regulates Tumor Growth by Promoting Angiogenesis
Scientific reports, 2017Co-Authors: Keiichi Asano, Courtney M. Nelson, Sumeda Nandadasa, Noriko Aramaki-hattori, Daniel J. Lindner, Tyler J. Alban, Junko Inagaki, Takashi Ohtsuki, Toshitaka Oohashi, Suneel S. ApteAbstract:The proteoglycan Versican is implicated in growth and metastases of several cancers. Here we investigated a potential contribution of stromal Versican to tumor growth and angiogenesis. We initially determined Versican expression by several cancer cell lines. Among these, MDA-MB231 and B16F10 had none to minimal expression in contrast to Lewis lung carcinoma (LLC). Notably, tumors arising from these cell lines had higher Versican levels than the cell lines themselves suggesting a contribution from the host-derived tumor stroma. In LLC-derived tumors, both the tumor and stroma expressed Versican at high levels. Thus, tumor stroma can make a significant contribution to tumor Versican content. Versican localized preferentially to the vicinity of tumor vasculature and macrophages in the tumor. However, an ADAMTS protease-generated Versican fragment uniquely localized to vascular endothelium. To specifically determine the impact of host/stroma-derived Versican we therefore compared growth of tumors from B16F10 cells, which produced littleVersican, in Vcan hdf/+ mice and wild-type littermates. Tumors in Vcan hdf/+ mice had reduced growth with a lower capillary density and accumulation of capillaries at the tumor periphery. These findings illustrate the variability of tumor cell line expression of Versican, and demonstrate that Versican is consistently contributed by the stromal tissue, where it contributes to tumor angiogenesis.
-
Isolation and purification of Versican and analysis of Versican proteolysis.
Methods in molecular biology (Clifton N.J.), 2014Co-Authors: Simon J. Foulcer, Anthony J. Day, Suneel S. ApteAbstract:Versican is a widely distributed chondroitin sulfate proteoglycan that forms large complexes with the glycosaminoglycan hyaluronan (HA). As a consequence of HA binding to its receptor CD44 and interactions of the Versican C-terminal globular (G3) domain with a variety of extracellular matrix proteins, Versican is a key component of well-defined networks in pericellular matrix and extracellular matrix. It is crucial for several developmental processes in the embryo and there is increasing interest in its roles in cancer and inflammation. Versican proteolysis by ADAMTS proteases is highly regulated, occurs at specific peptide bonds, and is relevant to several physiological and disease mechanisms. In this chapter, methods are described for the isolation and detection of intact and cleaved Versican in tissues using morphologic and biochemical techniques. These, together with the methodologies for purification and analysis of recombinant Versican and a Versican fragment provided here, are likely to facilitate further progress on the biology of Versican and its proteolysis.
-
Determinants of Versican-V1 Proteoglycan Processing by the Metalloproteinase ADAMTS5
The Journal of biological chemistry, 2014Co-Authors: Simon J. Foulcer, Dieter R. Zimmermann, María T. Dours-zimmermann, Courtney M. Nelson, Maritza V. Quintero, Balagurunathan Kuberan, J. Larkin, Suneel S. ApteAbstract:Proteolysis of the Glu(441)-Ala(442) bond in the glycosaminoglycan (GAG) β domain of the Versican-V1 variant by a disintegrin-like and metalloproteinase domain with thrombospondin type 1 motif (ADAMTS) proteases is required for proper embryo morphogenesis. However, the processing mechanism and the possibility of additional ADAMTS-cleaved processing sites are unknown. We demonstrate here that if Glu(441) is mutated, ADAMTS5 cleaves inefficiently at a proximate upstream site but normally does not cleave elsewhere within the GAGβ domain. Chondroitin sulfate (CS) modification of Versican is a prerequisite for cleavage at the Glu(441)-Ala(442) site, as demonstrated by reduced processing of CS-deficient or chondroitinase ABC-treated Versican-V1. Site-directed mutagenesis identified the N-terminal CS attachment sites Ser(507) and Ser(525) as essential for processing of the Glu(441)-Ala(442) bond by ADAMTS5. A construct including only these two GAG chains, but not downstream GAG attachment sites, was cleaved efficiently. Therefore, CS chain attachment to Ser(507) and Ser(525) is necessary and sufficient for Versican proteolysis by ADAMTS5. Mutagenesis of Glu(441) and an antibody to a peptide spanning Thr(432)-Gly(445) (i.e. containing the scissile bond) reduced Versican-V1 processing. ADAMTS5 lacking the C-terminal ancillary domain did not cleave Versican, and an ADAMTS5 ancillary domain construct bound Versican-V1 via the CS chains. We conclude that docking of ADAMTS5 with two N-terminal GAG chains of Versican-V1 via its ancillary domain is required for Versican processing at Glu(441)-Ala(442). V1 proteolysis by ADAMTS1 demonstrated a similar requirement for the N-terminal GAG chains and Glu(441). Therefore, Versican cleavage can be inhibited substantially by mutation of Glu(441), Ser(507), and Ser(525) or by an antibody to the region of the scissile bond.
-
the multiple complex roles of Versican and its proteolytic turnover by adamts proteases during embryogenesis
Matrix Biology, 2014Co-Authors: Sumeda Nandadasa, Simon J. Foulcer, Suneel S. ApteAbstract:Embryonic development is an exceptionally dynamic process, requiring a provisional extracellular matrix that is amenable to rapid remodeling, and proteolytic or non-proteolytic mechanisms that can remodel the major components of this matrix. Versican is a chondroitin-sulfate proteoglycan that forms highly hydrated complexes with hyaluronan and is widely distributed in the provisional matrix of mammalian embryos. It has been extensively studied in the context of cardiovascular morphogenesis, neural crest cell migration and skeletal development. Analysis of Vcan transgenic mice has established the requirement for Versican in cardiac development and its role in skeletogenesis. The ADAMTS family includes several Versican-degrading proteases that are active during remodeling of the embryonic provisional matrix, especially during sculpting of Versican-rich tissues. Versican is cleaved at specific peptide bonds by ADAMTS proteases, and the cleavage products are detectable by neo-epitope antibodies. Myocardial compaction, closure of the secondary palate (in which neural crest derived cells participate), endocardial cushion remodeling, myogenesis and interdigital web regression are developmental contexts in which ADAMTS-mediated Versican proteolysis has been identified as a crucial requirement. ADAMTS proteases are expressed coordinately and function cooperatively in many of these contexts. In addition to Versican clearance, ADAMTS proteases generate a bioactive Versican fragment containing the N-terminal G1 domain, which we have named versikine. This review promotes the view that the embryonic extracellular matrix has evolved not only to provide a permissive environment for embryo growth and morphogenesis, but through its dissolution to influence and regulate cellular processes.
-
pericellular Versican regulates the fibroblast myofibroblast transition a role for adamts5 protease mediated proteolysis
Journal of Biological Chemistry, 2011Co-Authors: Noriko Hattori, David A. Carrino, Courtney M. Nelson, James D. Wylie, Mark E Lauer, Amit Vasanji, Suneel S. ApteAbstract:The cell and its glycosaminoglycan-rich pericellular matrix (PCM) comprise a functional unit. Because modification of PCM influences cell behavior, we investigated molecular mechanisms that regulate PCM volume and composition. In fibroblasts and other cells, aggregates of hyaluronan and Versican are found in the PCM. Dermal fibroblasts from Adamts5−/− mice, which lack a Versican-degrading protease, ADAMTS5, had reduced Versican proteolysis, increased PCM, altered cell shape, enhanced α-smooth muscle actin (SMA) expression and increased contractility within three-dimensional collagen gels. The myofibroblast-like phenotype was associated with activation of TGFβ signaling. We tested the hypothesis that fibroblast-myofibroblast transition in Adamts5−/− cells resulted from Versican accumulation in PCM. First, we noted that Versican overexpression in human dermal fibroblasts led to increased SMA expression, enhanced contractility, and increased Smad2 phosphorylation. In contrast, dermal fibroblasts from Vcan haploinsufficient (Vcanhdf/+) mice had reduced contractility relative to wild type fibroblasts. Using a genetic approach to directly test if myofibroblast transition in Adamts5−/− cells resulted from increased PCM Versican content, we generated Adamts5−/−;Vcanhdf/+ mice and isolated their dermal fibroblasts for comparison with dermal fibroblasts from Adamts5−/− mice. In Adamts5−/− fibroblasts, Vcan haploinsufficiency or exogenous ADAMTS5 restored normal fibroblast contractility. These findings demonstrate that altering PCM Versican content through proteolytic activity of ADAMTS5 profoundly influenced the dermal fibroblast phenotype and may regulate a phenotypic continuum between the fibroblast and its alter ego, the myofibroblast. We propose that a physiological function of ADAMTS5 in dermal fibroblasts is to maintain optimal Versican content and PCM volume by continually trimming Versican in hyaluronan-Versican aggregates.
Albert Yee - One of the best experts on this subject based on the ideXlab platform.
-
Versican 3 untranslated region 3 utr promotes dermal wound repair and fibroblast migration by regulating mirna activity
Biochimica et Biophysica Acta, 2014Co-Authors: Weining Yang, Albert YeeAbstract:Versican is an extracellular chondroitin sulfate proteoglycan which functions as a structural molecule but can also regulate a variety of cellular activities. This study was designed to explore the roles of Versican in the process of dermal wound repair. To elevate levels of Versican, we ectopically expressed the Versican 3'-untranslated region (3'UTR) as a competitive endogenous RNA to modulate expression of Versican. We demonstrated that wounds closed faster in transgenic mice expressing the Versican 3'UTR, as compared to those in wildtype mice. We stably expressed Versican 3'UTR in NIH3T3 fibroblasts and found that the 3'UTR-transfected cells showed increased migratory capacity relative to vector-transfected cells. Interestingly, we found that the 3'UTRs of Versican and β-catenin shared common microRNAs (miRNAs) including miR-185, miR-203*, miR-690, miR-680, and miR-434-3p. Luciferase assays showed that all of these miRNAs could target the 3'UTRs of both Versican and β-catenin, when the luciferase constructs contained fragments harboring the miRNA binding sites. As a consequence, expression of both Versican and β-catenin was up-regulated, which was confirmed in vitro and in vivo. Transfection with small interfering RNAs (siRNAs) targeting the Versican 3'UTR abolished the 3'UTR's effects on cell migration and invasion. Taken together, these results demonstrate that Versican plays important roles in wound repair and that Versican messenger RNAs (mRNAs) could compete with endogenous RNAs for regulating miRNA functions.
-
Roles of Versican in cancer biology -tumorigenesis, progression and metastasis
Histology and histopathology, 2013Co-Authors: Weining Yang, Albert YeeAbstract:Versican, a large extracellular matrix proteoglycan accumulates in tumor stroma and plays a key role in both malignant transformation and tumor progression. Increased Versican expression has been observed in a wide range of malignant tumors, and has been associated with both cancer relapse and poor patient outcomes in breast, prostate, and many other cancer types. Through negatively-charged chondroitin and dermatan sulfate side chains or interactions of the G1 and G3 domains, Versican is able to regulate many cellular processes including cell adhesion, proliferation, apoptosis, migration, angiogenesis, invasion and metastasis. In this review, the biological roles that Versican plays in cancer development are presented. Therapeutic targeting of Versican in malignant tumors is also discussed.
-
Versican 3 untranslated region 3 utr functions as a cerna in inducing the development of hepatocellular carcinoma by regulating mirna activity
The FASEB Journal, 2013Co-Authors: Ling Fang, Weining Yang, Albert Yee, Xiangling Yang, Kui Chen, Anand Ghanekar, Gary A Levy, Jim W Xuan, Zhongli Gao, Feng XieAbstract:This study was designed to explore the role of Versican in the development of hepatocellular carcinoma (HCC). Ectopic expression of the Versican 3'-untranslated region (3'-UTR) was studied as a competitive endogenous RNA for regulating miRNA functions. We used this approach to modulate the expression of Versican and its related proteins in 3'-UTR transgenic mice and in the liver cancer cell line HepG2, stably transfected with the 3'-UTR or a control vector. We demonstrated that transgenic mice expressing the Versican 3'-UTR developed HCC and increased expression of Versican isoforms V0 and V1. HepG2 cells transfected with Versican 3'-UTR displayed increased proliferation, survival, migration, invasion, colony formation, and enhanced endothelial cell growth, but decreased apoptosis. We found that Versican 3'-UTR could bind to miRNAs miR-133a, miR-199a*, miR-144, and miR-431 and also interacted with CD34 and fibronectin. As a consequence, expression of Versican, CD34, and fibronectin was up-regulated by ectopic transfection of the Versican 3'-UTR, which was confirmed in HepG2 cells and in transgenic mice as compared with wild-type controls. Transfection with siRNAs targeting the Versican 3'-UTR abolished the effects of the 3'-UTR. Taken together, these results demonstrate that Versican V0 and V1 isoforms play important roles in HCC development and that Versican mRNAs compete with endogenous RNAs in regulating miRNA functions.
-
The Role of Versican in Modulating Breast Cancer Cell Self-renewal
Molecular cancer research : MCR, 2013Co-Authors: Ling Fang, Bing L. Yang, Yaou Zhang, Burton B Yang, Wang Sheng, Xiangling Yang, Arun Seth, Albert YeeAbstract:Versican is highly expressed during the early stages of tissue development and its expression is elevated during wound repair and tumor growth. There is little literature on the potential role of breast cancer stem cells on the cellular-extracellular matrix interactions involving Versican. An anti-Versican short hairpin RNA (shRNA) was used to observe the effect of reduction of Versican on breast cancer self-renewal. A Versican G3 construct was exogenously expressed in breast cancer cell lines. Colony formation and mammosphere formation assays were conducted; flow cytometry was applied to analyze the prevalence of side population cells. The Versican G3- and vector-transfected 66c14 cells were injected transdermally into BALB/c mice as a 10-fold dilution series from 1 × 10(5) to 1 × 10(2) cells per mouse. Versican G3 domain enhanced breast cancer self-renewal in both experimental in vitro and in vivo models. Versican G3-transfected cells contained high levels of side population cells, formed more mammospheres when cultured in the serum-free medium, and formed a greater number and larger colonies. Reduction of Versican's functionality through anti-Versican shRNA or knocking out the EGF-like motifs reduced the effect of Versican on enhancing mammosphere and colony formation. Versican-enhanced self-renewal played a role in enhanced chemotherapeutic drug resistance, relating partly to the upregulated expression of EGF receptor (EGFR) signaling. Versican is highly expressed in breast cancer progenitor cells and was maintained at high levels before cell differentiation. Overexpression of Versican enhanced breast cancer self-renewal through EGFR/AKT/GSK-3β (S9P) signaling and conferred resistant to chemotherapeutic drugs tested.
Bruce M Mcmanus - One of the best experts on this subject based on the ideXlab platform.
-
Versican localizes to the nucleus in proliferating mesenchymal cells.
Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology, 2015Co-Authors: Jon M Carthy, Thomas Abraham, Anna Meredith, Seti Boroomand, Bruce M McmanusAbstract:Abstract Objective Versican is a versatile and highly interactive chondroitin sulfate proteoglycan that is found in the extracellular matrix (ECM) of many tissues and is a major component of developing and developed lesions in atherosclerotic vascular disease. In this paper, we present data to indicate that Versican may have important intracellular functions in addition to its better known roles in the ECM. Methods and Results Rat aortic smooth muscle cells were fixed and immunostained for Versican and images of fluorescently labeled cells were obtained by confocal microscopy. Intracellular Versican was detected in the nucleus and cytosol of vascular smooth muscle cells. The use of a synthetic neutralizing peptide eliminated Versican immunostaining, demonstrating the specificity of the antibody used in this study. Western blot of pure nuclear extracts confirmed the presence of Versican in the nucleus, and multifluorescent immunostaining showed strong colocalization of Versican and nucleolin, suggesting a nucleolar localization of Versican in nondividing cells. In dividing valve interstitial cells, a strong signal for Versican was observed in and around the condensed chromosomes during the various stages of mitosis. Multifluorescent immunostaining for Versican and tubulin revealed Versican aggregated at opposing poles of the mitotic spindle during metaphase. Knockdown of Versican expression using siRNA disrupted the organization of the mitotic spindle and led to the formation of multipolar spindles during metaphase. Conclusions Collectively, these data suggest an intracellular function for Versican in vascular cells where it appears to play a role in mitotic spindle organization during cell division. These observations open a new avenue for studies of Versican, suggesting even more diverse roles in vascular health and disease.
-
Versican V1 Overexpression Induces a Myofibroblast-Like Phenotype in Cultured Fibroblasts.
PloS one, 2015Co-Authors: Jon M Carthy, Zongshu Luo, Thomas Abraham, Anna Meredith, Seti Boroomand, Darryl A. Knight, Bruce M McmanusAbstract:Background Versican, a chondroitin sulphate proteoglycan, is one of the key components of the provisional extracellular matrix expressed after injury. The current study evaluated the hypothesis that a Versican-rich matrix alters the phenotype of cultured fibroblasts. Methods and Results The full-length cDNA for the V1 isoform of human Versican was cloned and the recombinant proteoglycan was expressed in murine fibroblasts. Versican expression induced a marked change in fibroblast phenotype. Functionally, the Versican-expressing fibroblasts proliferated faster and displayed enhanced cell adhesion, but migrated slower than control cells. These changes in cell function were associated with greater N-cadherin and integrin β1 expression, along with increased FAK phosphorylation. The Versican-expressing fibroblasts also displayed expression of smooth muscle α-actin, a marker of myofibroblast differentiation. Consistent with this observation, the Versican fibroblasts displayed increased synthetic activity, as measured by collagen III mRNA expression, as well as a greater capacity to contract a collagen lattice. These changes appear to be mediated, at least in part, by an increase in active TGF-β signaling in the Versican expressing fibroblasts, and this was measured by phosphorylation and nuclear accumulation of SMAD2. Conclusions Collectively, these data indicate Versican expression induces a myofibroblast-like phenotype in cultured fibroblasts.
-
a role for Versican in the development of leiomyosarcoma
Journal of Biological Chemistry, 2014Co-Authors: Paul Keire, Steven L Bressler, Joan M Lemire, Badreddin Edris, Brian P Rubin, Maziar Rahmani, Bruce M Mcmanus, Matt Van De Rijn, Thomas N. WightAbstract:Leiomyosarcoma (LMS) is a mesenchymal cancer that occurs throughout the body. Although LMS is easily recognized histopathologically, the cause of the disease remains unknown. Versican, an extracellular matrix proteoglycan, increases in LMS. Microarray analyses of 80 LMSs and 24 leiomyomas showed a significant elevated expression of Versican in human LMS versus benign leiomyomas. To explore the importance of Versican in this smooth muscle cell tumor, we used Versican-directed siRNA to knock down Versican expression in a LMS human cell line, SK-LMS-1. Decreased Versican expression was accompanied by slower rates of LMS cell proliferation and migration, increased adhesion, and decreased accumulation of the extracellular matrix macromolecule hyaluronan. Addition of purified Versican to cells expressing Versican siRNA restored cell proliferation to the level of LMS controls, increased the pericellular coat and the retention of hyaluronan, and decreased cell adhesion in a dose-dependent manner. The presence of Versican was not only synergistic with hyaluronan in increasing cell proliferation, but the depletion of Versican decreased hyaluronan synthase expression and decreased the retention of hyaluronan. When LMS cells stably expressing Versican siRNA were injected into nude mice, the resulting tumors displayed significantly less Versican and hyaluronan staining, had lower volumes, and had reduced levels of mitosis as compared with controls. Collectively, these results suggest a role for using Versican as a point of control in the management and treatment of LMS.
-
Versican and CD44 in in vitro valvular interstitial cell injury and repair.
Cardiovascular pathology : the official journal of the Society for Cardiovascular Pathology, 2011Co-Authors: Jon M Carthy, Seti Boroomand, Bruce M McmanusAbstract:Abstract Background Versican is one of the key components of the extracellular matrix (ECM) that is expressed during injury, inflammatory, and repair processes. The current study evaluated the relationship between Versican and the membrane receptor CD44 during in vitro valvular interstitial cell (VIC) injury and repair. Methods Subconfluent, confluent, and wounded cultures of human VICs were fixed and immunostained to detect Versican and the membrane receptor CD44. To examine the relationship between Versican and CD44, a blocking antibody to CD44 was added to cultured VICs, and in vitro wound repair along with pericellular Versican organization and stress fiber formation were examined. Results Immunohistochemistry demonstrated that Versican is prominent intracellularly, as well as extracellularly, in actively proliferating VICs. In contrast, Versican was only localized to fibrils in the extracellular space in between cells in confluent (quiescent) cultures. Following wounding, Versican expression was up-regulated, and it was secreted as ECM at the trailing edge of migrating cells. The staining for CD44 was similarly localized to the trailing edge of migrating VICs in wounded cultures. Treatment of VICs with a CD44-blocking antibody disrupted the organization of Versican in the pericellular matrix and inhibited stress fiber formation in these cells. Functionally, blocking CD44 significantly inhibited VIC-mediated contraction of type I collagen gels (35.7%±0.7% vs. 23.3%±1.4% of initial gel area, P Conclusions Versican is a key component of the provisional wound repair ECM that is expressed following injury to VICs. The receptor CD44 plays an important role in organizing the provisional ECM. Summary Our data suggests VICs synthesize and secrete Versican following injury. These cells also up-regulate CD44, a receptor that binds Versican. Blocking CD44 disrupted the organization of Versican and inhibited stress fiber formation. Functionally, blocking CD44 inhibited cell-mediated contraction of a collagen matrix. Collectively, these data suggest Versican expression and organization are important to valve cell injury and repair.
-
Androgen Receptor Regulation of the Versican Gene through an Androgen Response Element in the Proximal Promoter
The Journal of biological chemistry, 2007Co-Authors: Jason T Read, Maziar Rahmani, Bruce M Mcmanus, Seti Boroomand, Sima Allahverdian, Paul S RennieAbstract:Abstract Versican, one of the key components of prostatic stroma, plays a central role in tumor initiation and progression. Here, we investigated promoter elements and mechanisms of androgen receptor (AR)-mediated regulation of the Versican gene in prostate cancer cells. Using transient transfection assays in prostate cancer LNCaP and cervical cancer HeLa cells engineered to express the AR, we demonstrate that the synthetic androgen R1881 and dihydrotestosterone stimulate expression of a Versican promoter-driven luciferase reporter vector (Versican-Luc). Further, both basal and androgen-stimulated Versican-Luc activities were significantly diminished in LNCaP cells, when AR gene expression was knocked down using a short hairpin RNA. Methylation-protection footprinting analysis revealed an AR-protected element between positions +75 and +102 of the proximal Versican promoter, which strongly resembled a consensus steroid receptor element. Electrophoretic mobility shift and supershift assays revealed strong and specific binding of the recombinant AR DNA binding domain to oligonucleotides corresponding to this protected DNA sequence. Site-directed mutagenesis of the steroid receptor element site markedly diminished R1881-stimulated Versican-Luc activity. In contrast to the response seen using LNCaP cells, R1881 did not significantly induce Versican promoter activity and mRNA levels in AR-positive prostate stromal fibroblasts. Interestingly, overexpression of β-catenin in the presence of androgen augmented Versican promoter activity 10- and 30-fold and enhanced Versican mRNA levels 2.8-fold in fibroblasts. In conclusion, we demonstrate that AR transactivates Versican expression, which may augment tumor-stromal interactions and may contribute to prostate cancer progression.
Burton B Yang - One of the best experts on this subject based on the ideXlab platform.
-
The Role of Versican in Modulating Breast Cancer Cell Self-renewal
Molecular cancer research : MCR, 2013Co-Authors: Ling Fang, Bing L. Yang, Yaou Zhang, Burton B Yang, Wang Sheng, Xiangling Yang, Arun Seth, Albert YeeAbstract:Versican is highly expressed during the early stages of tissue development and its expression is elevated during wound repair and tumor growth. There is little literature on the potential role of breast cancer stem cells on the cellular-extracellular matrix interactions involving Versican. An anti-Versican short hairpin RNA (shRNA) was used to observe the effect of reduction of Versican on breast cancer self-renewal. A Versican G3 construct was exogenously expressed in breast cancer cell lines. Colony formation and mammosphere formation assays were conducted; flow cytometry was applied to analyze the prevalence of side population cells. The Versican G3- and vector-transfected 66c14 cells were injected transdermally into BALB/c mice as a 10-fold dilution series from 1 × 10(5) to 1 × 10(2) cells per mouse. Versican G3 domain enhanced breast cancer self-renewal in both experimental in vitro and in vivo models. Versican G3-transfected cells contained high levels of side population cells, formed more mammospheres when cultured in the serum-free medium, and formed a greater number and larger colonies. Reduction of Versican's functionality through anti-Versican shRNA or knocking out the EGF-like motifs reduced the effect of Versican on enhancing mammosphere and colony formation. Versican-enhanced self-renewal played a role in enhanced chemotherapeutic drug resistance, relating partly to the upregulated expression of EGF receptor (EGFR) signaling. Versican is highly expressed in breast cancer progenitor cells and was maintained at high levels before cell differentiation. Overexpression of Versican enhanced breast cancer self-renewal through EGFR/AKT/GSK-3β (S9P) signaling and conferred resistant to chemotherapeutic drugs tested.
-
Versican modulates gap junction intercellular communication.
Journal of cellular physiology, 2007Co-Authors: Wang Sheng, Daniel Y. Lee, Haiheng Dong, Burton B YangAbstract:Versican is a large chondroitin sulfate proteoglycan and belongs to the family of lecticans. Versican possesses two globular domains, G1 and G3 domain, separated by a CS-attachment region. The CS-attachment region present in the middle region is divided into two spliced domains named CSalpha and beta. Alternative splicing of Versican generates at least four Versican isoforms named V0, V1, V2, and V3. We have successfully cloned the full-length cDNA of chick Versican isoforms V1 and V2 and found that Versican isoform V1 induced mesenchymal-epithelial transition in NIH3T3 cells. Mesenchymal-epithelial transition induced by V1 in NIH3T3 cells is characterized by expression of E-cadherin and occludin, two epithelial markers, and reduced expression of fibroblastic marker vimentin (Sheng et al., 2006, Mol Biol Cell. 17, 2009-2020). In the present studies, we found that Versican V1 isoform not only induced cell transition, but also increased intercellular communication via gap junction channels composed of connexin proteins. Our results showed that V1 induces plasma membrane localization of connexin 43, resulting in increased cell communication. This was further confirmed by blocking assays. Gap junctions mediated the transfer of small cytoplasmic molecules and the diffusion of second messenger molecules between adjacent cells. The ability of Versican in regulating gap junction implied a potential role of Versican in coordinating functions.
-
Versican Mediates Mesenchymal-Epithelial Transition
Molecular biology of the cell, 2006Co-Authors: Wang Sheng, Guizhi Wang, David P. La Pierre, Jianping Wen, Zhaoqun Deng, Chung-kwun Amy Wong, Daniel Y. Lee, Burton B YangAbstract:Versican is a large extracellular chondroitin sulfate proteoglycan that belongs to the family of lecticans. Alternative splicing of Versican generates at least four isoforms named V0, V1, V2, and V3. We show here that ectopic expression of Versican V1 isoform induced mesenchymal-epithelial transition (MET) in NIH3T3 fibroblasts, and inhibition of endogenous Versican expression abolished the MET in metanephric mesenchyme. MET in NIH3T3 cells was demonstrated by morphological changes and dramatic alterations in both membrane and cytoskeleton architecture. Molecular analysis showed that V1 promoted a "switch" in cadherin expression from N- to E-cadherin, resulting in epithelial specific adhesion junctions. V1 expression reduced vimentin levels and induced expression of occludin, an epithelial-specific marker, resulting in polarization of V1-transfected cells. Furthermore, an MSP (methylation-specific PCR) assay showed that N-cadherin expression was suppressed through methylation of its DNA promoter. Exogenous expression of N-cadherin in V1-transfected cells reversed V1's effect on cell aggregation. Reduction of E-cadherin expression by Snail transfection and siRNA targeting E-cadherin abolished V1-induced morphological alteration. Transfection of an siRNA construct targeting Versican also reversed the changed morphology induced by V1 expression. Silencing of endogenous Versican prevented MET of metanephric mesenchyme. Taken together, our results demonstrate the involvement of Versican in MET: expression of Versican is sufficient to induce MET in NIH3T3 fibroblasts and reduction of Versican expression decreased MET in metanephric mesenchyme.
-
Promotion of chondrocyte proliferation by Versican mediated by G1 domain and EGF-like motifs
Journal of cellular biochemistry, 1999Co-Authors: Yaou Zhang, Bing L. Yang, Chris Kiani, Liu Cao, Burton B YangAbstract:We have previously demonstrated that Versican stimulated NIH3T3 fibroblast proliferation. Since Versican is expressed in cartilage, we investigated whether Versican plays a role in chondrocyte proliferation. We developed a technique to stably express a recombinant Versican mini-gene in chicken chondrocytes, and its effect on chondrocyte proliferation was analyzed by the increase in cell number. The effect of cell adhesion on cell proliferation was tested. Finally, the Versican mini-gene was truncated to assess the role of EGF-like motifs in cell proliferation. Expression of the recombinant Versican mini-gene stimulated chondrocyte proliferation. Antisense oligonucleotides complementary to Versican inhibited chondrocyte proliferation. The G1 domain of Versican stimulated chondrocyte proliferation by destabilizing chondrocyte adhesion. Furthermore, deletion of the two EGF-like motifs from the G3 domain also reduced the function of Versican in stimulating cell proliferation. Versican enhances chondrocyte proliferation through a mechanism involving its G1 and G3 domains. This finding may have implications for our understanding of the pathogenesis of various joint diseases. J. Cell. Biochem. 73:445–457, 1999. © 1999 Wiley-Liss, Inc.
-
Versican enhances locomotion of astrocytoma cells and reduces cell adhesion through its G1 domain
Journal of Neuropathology and Experimental Neurology, 1999Co-Authors: Yaou Zhang, Bing L. Yang, Chris Kiani, Katherine S Allan, Beverley Young, Burton B YangAbstract:Versican is a large extracellular proteoglycan and is expressed in a variety of tissues including the central nervous system. A malignant astrocytoma cell line U87 with high motility expressed a higher level of Versican than another malignant astrocytoma cell line U343 with lower motility. We observed that the U87 cells were less adherent to tissue culture plates than the U343 cells. To investigate the role of Versican in astrocytoma cell migration, we generated recombinant products of a mini-Versican construct expressed in COS-7 cells. We found that the mini-Versican products enhanced astrocytoma cell migration. Furthermore, enhanced migration was promoted by the G1 domain but not the G3 domain of Versican. We introduced culture medium containing products of the mini-Versican, the G1, and the G3 constructs separately into the astrocytoma cell lines U87 and U343. The mini-Versican and the G1 construct, but not the G3 construct, were shown to reduce astrocytoma cell adhesion. The present data suggest that Versican exerts its effect on astrocytoma cell migration and adhesion through the G1 domain.
