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Renato V. Iozzo - One of the best experts on this subject based on the ideXlab platform.
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A current view of Perlecan in physiology and pathology: A mosaic of functions.
Matrix biology : journal of the International Society for Matrix Biology, 2016Co-Authors: Maria A Gubbiotti, Thomas Neill, Renato V. IozzoAbstract:Perlecan, a large basement membrane heparan sulfate proteoglycan, is expressed in a wide array of tissues where it regulates diverse cellular processes including bone formation, inflammation, cardiac development, and angiogenesis. Here we provide a contemporary review germane to the biology of Perlecan encompassing its genetic regulation as well as an analysis of its modular protein structure as it pertains to function. As Perlecan signaling from the extracellular matrix converges on master regulators of autophagy, including AMPK and mTOR, via a specific interaction with vascular endothelial growth factor receptor 2, we specifically focus on the mechanism of action of Perlecan in autophagy and angiogenesis and contrast the role of endorepellin, the C-terminal fragment of Perlecan, in these cellular and morphogenic events.
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the role of vascular derived Perlecan in modulating cell adhesion proliferation and growth factor signaling
Matrix Biology, 2014Co-Authors: Megan S. Lord, James Melrose, Renato V. Iozzo, Christine Y Chuang, Michael J Davies, John M. WhitelockAbstract:Smooth muscle cell proliferation can be inhibited by heparan sulfate proteoglycans whereas the removal or digestion of heparan sulfate from Perlecan promotes their proliferation. In this study we characterized the glycosaminoglycan side chains of Perlecan isolated from either primary human coronary artery smooth muscle or endothelial cells and determined their roles in mediating cell adhesion and proliferation, and in fibroblast growth factor (FGF) binding and signaling. Smooth muscle cell Perlecan was decorated with both heparan sulfate and chondroitin sulfate, whereas endothelial Perlecan contained exclusively heparan sulfate chains. Smooth muscle cells bound to the protein core of Perlecan only when the glycosaminoglycans were removed, and this binding involved a novel site in domain III as well as domain V/endorepellin and the α2β1 integrin. In contrast, endothelial cells adhered to the protein core of Perlecan in the presence of glycosaminoglycans. Smooth muscle cell Perlecan bound both FGF1 and FGF2 via its heparan sulfate chains and promoted the signaling of FGF2 but not FGF1. Also endothelial cell Perlecan bound both FGF1 and FGF2 via its heparan sulfate chains, but in contrast, promoted the signaling of both growth factors. Based on this differential bioactivity, we propose that Perlecan synthesized by smooth muscle cells differs from that synthesized by endothelial cells by possessing different signaling capabilities, primarily, but not exclusively, due to a differential glycanation. The end result is a differential modulation of cell adhesion, proliferation and growth factor signaling in these two key cellular constituents of blood vessels.
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heparan sulfate dependent signaling of fibroblast growth factor 18 by chondrocyte derived Perlecan
Biochemistry, 2010Co-Authors: Christine Y Chuang, Sarah M. Knox, James Melrose, Renato V. Iozzo, Megan S. Lord, Martin D Rees, Craig Freeman, John M. WhitelockAbstract:Perlecan is a large multidomain proteoglycan that is essential for normal cartilage development. In this study, Perlecan was localized in the pericellular matrix of hypertrophic chondrocytes in developing human cartilage rudiments. Perlecan immunopurified from medium conditioned by cultured human fetal chondrocytes was found to be substituted with heparan sulfate (HS), chondroitin sulfate (CS), and keratan sulfate (KS). Ligand and carbohydrate engagement (LACE) assays demonstrated that immunopurified chondrocyte-derived Perlecan formed HS-dependent ternary complexes with fibroblast growth factor (FGF) 2 and either FGF receptors (FGFRs) 1 or 3; however, these complexes were not biologically active in the BaF32 cell system. Chondrocyte-derived Perlecan also formed HS-dependent ternary complexes with FGF18 and FGFR3. The proliferation of BaF32 cells expressing FGFR3 was promoted by chondrocyte-derived Perlecan in the presence of FGF18, and this activity was reduced by digestion of the HS with either heparina...
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Perlecan regulates developmental angiogenesis by modulating the VEGF-VEGFR2 axis.
Matrix Biology, 2009Co-Authors: Jason J. Zoeller, John M. Whitelock, Renato V. IozzoAbstract:Abstract Using the zebrafish, we previously identified a central function for Perlecan during angiogenic blood vessel development. Here, we explored the nature of Perlecan function during developmental angiogenesis. A close examination of individual endothelial cell behavior revealed that Perlecan is required for proper endothelial cell migration and proliferation. Because these events are largely mediated by VEGF-VEGFR2 signaling, we investigated the relationship between Perlecan and the VEGF pathway. We discovered that Perlecan knockdown caused an abnormal increase and redistribution of total VEGF-A protein suggesting that Perlecan is required for the appropriate localization of VEGF-A. Importantly, we linked Perlecan function to the VEGF pathway by efficiently rescuing the Perlecan morphant phenotype by microinjecting VEGF-A165 protein or mRNA. Combining the strategic localization of Perlecan throughout the vascular basement membrane along with its growth factor-binding ability, we hypothesized a major role for Perlecan during the establishment of the VEGF gradient which provides the instructive cues to endothelial cells during angiogenesis. In support of this hypothesis we demonstrated that human Perlecan bound in a heparan sulfate-dependent fashion to VEGF-A165. Moreover, Perlecan enhanced VEGF mediated VEGFR2 activation of human endothelial cells. Collectively, our results indicate that Perlecan coordinates developmental angiogenesis through modulation of VEGF-VEGFR2 signaling events. The identification of angiogenic factors, such as Perlecan, and their role in vertebrate development will not only enhance overall understanding of the molecular basis of angiogenesis, but may also provide new insight into angiogenesis-based therapeutic approaches.
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Novel interactions of Perlecan: unraveling Perlecan's role in angiogenesis.
Microscopy Research and Technique, 2008Co-Authors: Gregory J. Bix, Renato V. IozzoAbstract:Perlecan, a highly conserved and ubiquitous basement membrane heparan sulfate proteoglycan, is essential for life, inasmuch as its absence results in embryonic lethality in mice and C. elegans, and neonatal lethality in humans. Perlecan plays an essential role in vasculogenesis and chondrogenesis, as well as in pathological states where these processes are maladapted. Although a large body of evidence supports a pro-angiogenic role for Perlecan, recent findings suggests that portions of the Perlecan protein core can be antiangiogenic, requiring a further evaluation of the functioning of this complex molecule. This review is focused on the genetics of mammalian and nonmammalian Perlecan, the elucidation of its novel interacting partners and its role in angiogenesis. By more fully understanding Perlecan's functioning in angiogenesis, we may gain invaluable insight that could lead to therapeutic interventions in cancer and other pathologic states. Microsc. Res. Tech., 2008. © 2008 Wiley-Liss, Inc.
John M. Whitelock - One of the best experts on this subject based on the ideXlab platform.
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Perlecan expression influences the keratin 15 positive cell population fate in the epidermis of aging skin
Aging (Albany NY), 2016Co-Authors: Morgan Dos Santos, John M. Whitelock, Anna Michopoulou, Valerie Andrefrei, Sophie Boulesteix, Christine Guicher, Guila Dayan, Odile Damour, Patricia RousselleAbstract:The epidermis is continuously renewed by stem cell proliferation and differentiation. Basal keratinocytes append the dermal-epidermal junction, a cell surface-associated, extracellular matrix that provides structural support and influences their behaviour. It consists of laminins, type IV collagen, nidogens, and Perlecan, which are necessary for tissue organization and structural integrity. Perlecan is a heparan sulfate proteoglycan known to be involved in keratinocyte survival and differentiation. Aging affects the dermal epidermal junction resulting in decreased contact with keratinocytes, thus impacting epidermal renewal and homeostasis. We found that Perlecan expression decreased during chronological skin aging. Our in vitro studies revealed reduced Perlecan transcript levels in aged keratinocytes. The production of in vitro skin models revealed that aged keratinocytes formed a thin and poorly organized epidermis. Supplementing these models with purified Perlecan reversed the phenomenon allowing restoration of a well-differentiated multi-layered epithelium. Perlecan down-regulation in cultured keratinocytes caused depletion of the cell population that expressed keratin 15. This phenomenon depended on the Perlecan heparan sulphate moieties, which suggested the involvement of a growth factor. Finally, we found defects in keratin 15 expression in the epidermis of aging skin. This study highlighted a new role for Perlecan in maintaining the self-renewal capacity of basal keratinocytes.
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the role of vascular derived Perlecan in modulating cell adhesion proliferation and growth factor signaling
Matrix Biology, 2014Co-Authors: Megan S. Lord, James Melrose, Renato V. Iozzo, Christine Y Chuang, Michael J Davies, John M. WhitelockAbstract:Smooth muscle cell proliferation can be inhibited by heparan sulfate proteoglycans whereas the removal or digestion of heparan sulfate from Perlecan promotes their proliferation. In this study we characterized the glycosaminoglycan side chains of Perlecan isolated from either primary human coronary artery smooth muscle or endothelial cells and determined their roles in mediating cell adhesion and proliferation, and in fibroblast growth factor (FGF) binding and signaling. Smooth muscle cell Perlecan was decorated with both heparan sulfate and chondroitin sulfate, whereas endothelial Perlecan contained exclusively heparan sulfate chains. Smooth muscle cells bound to the protein core of Perlecan only when the glycosaminoglycans were removed, and this binding involved a novel site in domain III as well as domain V/endorepellin and the α2β1 integrin. In contrast, endothelial cells adhered to the protein core of Perlecan in the presence of glycosaminoglycans. Smooth muscle cell Perlecan bound both FGF1 and FGF2 via its heparan sulfate chains and promoted the signaling of FGF2 but not FGF1. Also endothelial cell Perlecan bound both FGF1 and FGF2 via its heparan sulfate chains, but in contrast, promoted the signaling of both growth factors. Based on this differential bioactivity, we propose that Perlecan synthesized by smooth muscle cells differs from that synthesized by endothelial cells by possessing different signaling capabilities, primarily, but not exclusively, due to a differential glycanation. The end result is a differential modulation of cell adhesion, proliferation and growth factor signaling in these two key cellular constituents of blood vessels.
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Colocalization in vivo and association in vitro of Perlecan and elastin.
Histochemistry and Cell Biology, 2011Co-Authors: Anthony James Hayes, John M. Whitelock, Megan S. Lord, Susan M. Smith, Margaret M. Smith, Anthony S. Weiss, James MelroseAbstract:We have colocalized elastin and fibrillin-1 with Perlecan in extracellular matrix of tensional and weight-bearing connective tissues. Elastin and fibrillin-1 were identified as prominent components of paraspinal blood vessels, and posterior longitudinal ligament in the human fetal spine and outer annulus fibrosus of the fetal intervertebral disc. We also colocalized Perlecan with a synovial elastic basal lamina, where the attached synovial cells were observed to produce Perlecan. Elastin, fibrillin-1 and Perlecan were co-localized in the intima and media of small blood vessels in the synovium and in human fetal paraspinal blood vessels. Elastic fibers were observed at the insertion point of the anterior cruciate ligament to bone in the ovine stifle joint where they colocalized with Perlecan. Elastin has not previously been reported to be spatially associated with Perlecan in these tissues. Interactions between the tropoelastin and Perlecan heparan sulfate chains were demonstrated using quartz crystal microbalance with dissipation solid phase binding studies. Electrostatic interactions through the heparan sulfate chains of Perlecan and core protein mediated the interactions with tropoelastin, and were both important in the coacervation of tropoelastin and deposition of elastin onto Perlecan immobilized on the chip surface. This may help us to understand the interactions which are expected to occur in vivo between the tropoelastin and Perlecan to facilitate the deposition of elastin and formation of elastic microfibrils in situ and would be consistent with the observed distributions of these components in a number of connective tissues.
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peroxynitrite modifies the structure and function of the extracellular matrix proteoglycan Perlecan by reaction with both the protein core and the heparan sulfate chains
Free Radical Biology and Medicine, 2010Co-Authors: Eleanor C Kennett, John M. Whitelock, Michael J Davies, Martin D Rees, Ernst Malle, Astrid HammerAbstract:The heparan sulfate (HS) proteoglycan Perlecan is a major component of basement membranes, plays a key role in extracellular matrix (ECM) structure, interacts with growth factors and adhesion molecules, and regulates the adhesion, differentiation and proliferation of vascular cells. Atherosclerosis is characterized by chronic inflammation and the presence of oxidized materials within lesions, with the majority of protein damage present on ECM, rather than cell, proteins. Weakening of ECM structure plays a key role in lesion rupture, the major cause of heart attacks and strokes. In this study peroxynitrite, a putative lesion oxidant, is shown to damage Perlecan structurally and functionally. Exposure of human Perlecan to peroxynitrite decreases recognition by antibodies raised against both the core protein and heparan sulfate chains; dose-dependent formation of 3-nitrotyrosine was also detected. These effects were modulated by bicarbonate and reaction pH. Oxidant exposure resulted in aggregate formation, consistent with oxidative protein crosslinking. Peroxynitrite treatment modified functional properties of Perlecan that are dependent on both the protein core (decreased binding of human coronary artery endothelial cells), and the HS chains (diminished fibroblast growth factor-2 (FGF-2) receptor-mediated proliferation of Baf-32 cells). The latter is consistent with a decrease in FGF-2 binding to the HS chains of modified Perlecan. Immunofluorescence of advanced human atherosclerotic lesions provided evidence for the presence of Perlecan and extensive formation of 3-nitrotyrosine epitopes within the intimal region; these materials showing marked co-localization. These data indicate that peroxynitrite induces major structural and functional changes to Perlecan and that damage to this material occurs within human atherosclerotic lesions.
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heparan sulfate dependent signaling of fibroblast growth factor 18 by chondrocyte derived Perlecan
Biochemistry, 2010Co-Authors: Christine Y Chuang, Sarah M. Knox, James Melrose, Renato V. Iozzo, Megan S. Lord, Martin D Rees, Craig Freeman, John M. WhitelockAbstract:Perlecan is a large multidomain proteoglycan that is essential for normal cartilage development. In this study, Perlecan was localized in the pericellular matrix of hypertrophic chondrocytes in developing human cartilage rudiments. Perlecan immunopurified from medium conditioned by cultured human fetal chondrocytes was found to be substituted with heparan sulfate (HS), chondroitin sulfate (CS), and keratan sulfate (KS). Ligand and carbohydrate engagement (LACE) assays demonstrated that immunopurified chondrocyte-derived Perlecan formed HS-dependent ternary complexes with fibroblast growth factor (FGF) 2 and either FGF receptors (FGFRs) 1 or 3; however, these complexes were not biologically active in the BaF32 cell system. Chondrocyte-derived Perlecan also formed HS-dependent ternary complexes with FGF18 and FGFR3. The proliferation of BaF32 cells expressing FGFR3 was promoted by chondrocyte-derived Perlecan in the presence of FGF18, and this activity was reduced by digestion of the HS with either heparina...
John R. Hassell - One of the best experts on this subject based on the ideXlab platform.
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Perlecan modulates VEGF signaling and is essential for vascularization in endochondral bone formation
Matrix Biology, 2012Co-Authors: Muneaki Ishijima, Eri Arikawa-hirasawa, John R. Hassell, Nobuharu Suzuki, Kentaro Hozumi, Tomoya Matsunobu, Keisuke Kosaki, Haruka Kaneko, Yoshihiko YamadaAbstract:Perlecan (Hspg2) is a heparan sulfate proteoglycan expressed in basement membranes and cartilage. Perlecan deficiency (Hspg2−/−) in mice and humans causes lethal chondrodysplasia, which indicates that Perlecan is essential for cartilage development. However, the function of Perlecan in endochondral ossification is not clear. Here, we report the critical role of Perlecan in VEGF signaling and angiogenesis in growth plate formation. The Hspg2−/− growth plate was significantly wider but shorter due to severely impaired endochondral bone formation. Hypertrophic chondrocytes were differentiated in Hspg2−/− growth plates; however, removal of the hypertrophic matrix and calcified cartilage was inhibited. Although the expression of MMP-13, CTGF, and VEGFA was significantly upregulated in Hspg2−/− growth plates, vascular invasion into the hypertrophic zone was impaired, which resulted in an almost complete lack of bone marrow and trabecular bone. We demonstrated that cartilage Perlecan promoted activation of VEGF/VEGFR by binding to the VEGFR of endothelial cells. Expression of the Perlecan transgene specific to the cartilage of Hspg2−/− mice rescued their perinatal lethality and growth plate abnormalities, and vascularization into the growth plate was restored, indicating that Perlecan in the growth plate, not in endothelial cells, is critical in this process. These results suggest that Perlecan in cartilage is required for activating VEGFR signaling of endothelial cells for vascular invasion and for osteoblast migration into the growth plate. Thus, Perlecan in cartilage plays a critical role in endochondral bone formation by promoting angiogenesis essential for cartilage matrix remodeling and subsequent endochondral bone formation.
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The core protein of growth plate Perlecan binds FGF-18 and alters its mitogenic effect on chondrocytes
Archives of Biochemistry and Biophysics, 2007Co-Authors: Simone M. Smith, Leigh A. West, John R. HassellAbstract:Abstract Fibroblast growth factor-18 (FGF-18) has been shown to regulate the growth plate chondrocyte proliferation, hypertrophy and cartilage vascularization necessary for endochondral ossification. The heparan sulfate proteoglycan Perlecan is also critical for growth plate chondrocyte proliferation. FGF-18 null mice exhibit a skeletal dwarfism similar to that of Perlecan null mice. Growth plate Perlecan contains chondroitin sulfate (CS) and heparan sulfate (HS) chains and FGF-18 is known to bind to heparin and to heparan sulfate from some sources. We used cationic filtration and immunoprecipitation assays to investigate the binding of FGF-18 to Perlecan purified from the growth plate and to recombinant Perlecan domains expressed in COS-7 cells. FGF-18 bound to Perlecan with a Kd of 145 nM. Near saturation, ∼103 molecules of FGF-18 bound per molecule of Perlecan. At the lower concentrations used, FGF-18 bound with a Kd of 27.8 nM. This binding was not significantly altered by chondroitinase nor heparitinase digestion of Perlecan, but was substantially and significantly reduced by reduction and alkylation of the Perlecan core protein. This indicates that the Perlecan core protein (and not the CS nor HS chains) is involved in FGF-18 binding. FGF-18 bound equally to full-length Perlecan purified from the growth plate and to recombinant domains I–III and III of Perlecan. These data indicate that low affinity binding sites for FGF-18 are present in cysteine-rich regions of domain III of Perlecan. FGF-18 stimulated 3H-thymidine incorporation in growth plate chondrocyte cultures derived from the lower and upper proliferating zones by 9- and 14-fold, respectively. The addition of Perlecan reversed this increased incorporation in the lower proliferating chondrocytes by 74% and in the upper proliferating cells by 37%. These results suggest that Perlecan can bind FGF-18 and alter the mitogenic effect of FGF-18 on growth plate chondrocytes.
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heparan and chondroitin sulfate on growth plate Perlecan mediate binding and delivery of fgf 2 to fgf receptors
Matrix Biology, 2007Co-Authors: Simone M. Smith, Leigh A. West, John R. Hassell, Prasanthi Govindraj, Xiuqin Zhang, David M OrnitzAbstract:Fibroblast growth factor (FGF)-2 regulates chondrocyte proliferation in the growth plate. Heparan sulfate (HS) proteoglycans bind FGF-2. Perlecan, a heparan sulfate proteoglycan (HSPG) in the developing growth plate, however, contains both HS and chondroitin sulfate (CS) chains. The binding of FGF-2 to Perlecan isolated from the growth plate was evaluated using cationic filtration (CAF) and immunoprecipitation (IP) assays. FGF-2 bound to Perlecan in both the CAF and IP assays primarily via the HS chains on Perlecan. A maximum of 123 molecules of FGF-2 was calculated to bind per molecule of Perlecan. When digested with chondroitinase ABC to remove its CS chains, Perlecan augmented binding of FGF-2 to the FGFR-1 and FGFR-3 receptors and also increased FGF-2 stimulation of [ 3 H]-thymidine incorporation in BaF3 cells expressing these FGF receptors. These data show that growth plate Perlecan binds to FGF-2 by its HS chains but can only deliver FGF-2 to FGF receptors when its CS chains are removed.
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Changes in Perlecan during chondrocyte differentiation in the fetal bovine rib growth plate.
Journal of Orthopaedic Research, 2006Co-Authors: Leigh A. West, Prasanthi Govindraj, Thomas J. Koob, John R. HassellAbstract:Perlecan is a heparan sulfate proteoglycan present in the growth plate and essential for endochondral ossification. We evaluated the synthesis and structure of Perlecan in the different zones of the growth plate. The growth plates from fetal bovine ribs were isolated and sequentially sliced into 1-mm sections containing the hypertrophic zone, lower proliferative zone, upper proliferative zone, intermediate zone, and resting zone, respectively. The slices were then either incubated in culture medium with 35SO4 to measure total sulfated proteoglycan synthesis and Perlecan synthesis, extracted for Perlecan core protein analysis by Western blot, or extracted for Perlecan isolation and subsequent characterization of glycosaminoglycan size and disaccharide composition. 35SO4 incorporation into Perlecan was three–fourfold higher in the proliferating/hypertrophic zone than the resting zone. Western blot showed Perlecan content was greatest in the lower and upper proliferating zones and that a Perlecan fragment lacking portions of the N- and C-terminal domains containing heparan sulfate was also present in all zones. Purified Perlecan from the hypertrophic/lower proliferative zone had larger chondroitin sulfate chains and a different composition of CS and HS disaccharides than the Perlecan isolated from the resting zone. These results indicate Perlecan deposition is increased and is turned over during proliferation to be replaced by a Perlecan with a different sulfation pattern. © 2006 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 24:1317–1326, 2006
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structural and functional mutations of the Perlecan gene cause schwartz jampel syndrome with myotonic myopathy and chondrodysplasia
American Journal of Human Genetics, 2002Co-Authors: Eri Arikawahirasawa, John R. Hassell, Prasanthi Govindraj, Ichizo Nishino, Ikuya Nonaka, Clair A Francomano, Joseph M Devaney, Jurgen W Spranger, Roger E Stevenson, Susan T IannacconeAbstract:Perlecan, a large heparan sulfate proteoglycan, is a component of the basement membrane and other extracellular matrices and has been implicated in multiple biological functions. Mutations in the Perlecan gene (HSPG2) cause two classes of skeletal disorders: the relatively mild Schwartz-Jampel syndrome (SJS) and severe neonatal lethal dyssegmental dysplasia, Silverman-Handmaker type (DDSH). SJS is an autosomal recessive skeletal dysplasia characterized by varying degrees of myotonia and chondrodysplasia, and patients with SJS survive. The molecular mechanism underlying the chondrodystrophic myotonia phenotype of SJS is unknown. In the present report, we identify five different mutations that resulted in various forms of Perlecan in three unrelated patients with SJS. Heterozygous mutations in two patients with SJS either produced truncated Perlecan that lacked domain V or significantly reduced levels of wild-type Perlecan. The third patient had a homozygous 7-kb deletion that resulted in reduced amounts of nearly full-length Perlecan. Unlike DDSH, the SJS mutations result in different forms of Perlecan in reduced levels that are secreted to the extracellular matrix and are likely partially functional. These findings suggest that Perlecan has an important role in neuromuscular function and cartilage formation, and they define the molecular basis involved in the difference in the phenotypic severity between DDSH and SJS.
Eri Arikawahirasawa - One of the best experts on this subject based on the ideXlab platform.
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Perlecan a heparan sulfate proteoglycan regulates systemic metabolism with dynamic changes in adipose tissue and skeletal muscle
Scientific Reports, 2018Co-Authors: Yuri Yamashita, Satoshi Nakada, Toshinori Yoshihara, Takeshi Nara, Norihiko Furuya, Takashi Miida, Nobutaka Hattori, Eri ArikawahirasawaAbstract:Perlecan (HSPG2), a heparan sulfate proteoglycan, is a component of basement membranes and participates in a variety of biological activities. Here, we show physiological roles of Perlecan in both obesity and the onset of metabolic syndrome. The perinatal lethality-rescued Perlecan knockout (Hspg2-/--Tg) mice showed a smaller mass and cell size of white adipose tissues than control (WT-Tg) mice. Abnormal lipid deposition, such as fatty liver, was not detected in the Hspg2-/--Tg mice, and those mice also consumed more fat as an energy source, likely due to their activated fatty acid oxidation. In addition, the Hspg2-/--Tg mice demonstrated increased insulin sensitivity. Molecular analysis revealed the significantly relatively increased amount of the muscle fiber type IIA (X) isoform and a larger quantity of mitochondria in the skeletal muscle of Hspg2-/--Tg mice. Furthermore, the Perlecan-deficient skeletal muscle also had elevated levels of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α) protein. PGC1α expression is activated by exercise, and induces mitochondrial biosynthesis. Thus, Perlecan may act as a mechano-regulator of catabolism of both lipids and glucose by shifting the muscle fiber composition to oxidative fibers. Our data suggest that downregulation of Perlecan is a promising strategy to control metabolic syndrome.
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structural and functional mutations of the Perlecan gene cause schwartz jampel syndrome with myotonic myopathy and chondrodysplasia
American Journal of Human Genetics, 2002Co-Authors: Eri Arikawahirasawa, John R. Hassell, Prasanthi Govindraj, Ichizo Nishino, Ikuya Nonaka, Clair A Francomano, Joseph M Devaney, Jurgen W Spranger, Roger E Stevenson, Susan T IannacconeAbstract:Perlecan, a large heparan sulfate proteoglycan, is a component of the basement membrane and other extracellular matrices and has been implicated in multiple biological functions. Mutations in the Perlecan gene (HSPG2) cause two classes of skeletal disorders: the relatively mild Schwartz-Jampel syndrome (SJS) and severe neonatal lethal dyssegmental dysplasia, Silverman-Handmaker type (DDSH). SJS is an autosomal recessive skeletal dysplasia characterized by varying degrees of myotonia and chondrodysplasia, and patients with SJS survive. The molecular mechanism underlying the chondrodystrophic myotonia phenotype of SJS is unknown. In the present report, we identify five different mutations that resulted in various forms of Perlecan in three unrelated patients with SJS. Heterozygous mutations in two patients with SJS either produced truncated Perlecan that lacked domain V or significantly reduced levels of wild-type Perlecan. The third patient had a homozygous 7-kb deletion that resulted in reduced amounts of nearly full-length Perlecan. Unlike DDSH, the SJS mutations result in different forms of Perlecan in reduced levels that are secreted to the extracellular matrix and are likely partially functional. These findings suggest that Perlecan has an important role in neuromuscular function and cartilage formation, and they define the molecular basis involved in the difference in the phenotypic severity between DDSH and SJS.
Yoshihiko Yamada - One of the best experts on this subject based on the ideXlab platform.
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abstract wmp117 Perlecan regulates pericyte dynamics in the repair process of the blood brain barrier against ischemic stroke
Stroke, 2018Co-Authors: Kuniyuki Nakamura, Tomoko Ikeuchi, Peipei Zhang, Craig Rhodes, Yuta Chiba, Tetsuro Ago, Yohsuke Mukouyama, Yoshihiko YamadaAbstract:Introduction: The blood-brain barrier (BBB) breakdown occurs when the integrity of BBB components is lost as a consequence of ischemic stroke. Perlecan, a major heparan sulfate proteoglycan of basement membranes, is expressed by endothelial cells (ECs) and is adjacent to pericytes (PCs), suggesting supportive functions in the BBB. Recent studies highlight the importance of PCs in the process of repairing BBB functions, which are triggered by the upregulation of platelet-derived growth factor receptor β (PDGFRβ). We hypothesized that Perlecan may play a protective role in BBB maintenance through the interaction with PCs during the repair process of the BBB disruption. Methods: We induced a 60-minute middle cerebral artery occlusion (MCAO) in adult conditional Perlecan-deficient (Perlecan-/--Tg) mice in a C57BL/6 background, which express the Perlecan transgene only in cartilage to rescue the perinatal lethality of Perlecan-/- mice. Recombinant Perlecan C-terminal domain V (DV) was used for in vitro assays....
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Perlecan modulates VEGF signaling and is essential for vascularization in endochondral bone formation
Matrix Biology, 2012Co-Authors: Muneaki Ishijima, Eri Arikawa-hirasawa, John R. Hassell, Nobuharu Suzuki, Kentaro Hozumi, Tomoya Matsunobu, Keisuke Kosaki, Haruka Kaneko, Yoshihiko YamadaAbstract:Perlecan (Hspg2) is a heparan sulfate proteoglycan expressed in basement membranes and cartilage. Perlecan deficiency (Hspg2−/−) in mice and humans causes lethal chondrodysplasia, which indicates that Perlecan is essential for cartilage development. However, the function of Perlecan in endochondral ossification is not clear. Here, we report the critical role of Perlecan in VEGF signaling and angiogenesis in growth plate formation. The Hspg2−/− growth plate was significantly wider but shorter due to severely impaired endochondral bone formation. Hypertrophic chondrocytes were differentiated in Hspg2−/− growth plates; however, removal of the hypertrophic matrix and calcified cartilage was inhibited. Although the expression of MMP-13, CTGF, and VEGFA was significantly upregulated in Hspg2−/− growth plates, vascular invasion into the hypertrophic zone was impaired, which resulted in an almost complete lack of bone marrow and trabecular bone. We demonstrated that cartilage Perlecan promoted activation of VEGF/VEGFR by binding to the VEGFR of endothelial cells. Expression of the Perlecan transgene specific to the cartilage of Hspg2−/− mice rescued their perinatal lethality and growth plate abnormalities, and vascularization into the growth plate was restored, indicating that Perlecan in the growth plate, not in endothelial cells, is critical in this process. These results suggest that Perlecan in cartilage is required for activating VEGFR signaling of endothelial cells for vascular invasion and for osteoblast migration into the growth plate. Thus, Perlecan in cartilage plays a critical role in endochondral bone formation by promoting angiogenesis essential for cartilage matrix remodeling and subsequent endochondral bone formation.
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morphogenetic roles of Perlecan in the tooth enamel organ an analysis of overexpression using transgenic mice
Matrix Biology, 2011Co-Authors: Hiroko Idayonemochi, Yoshihiko Yamada, Ichiro Satokata, Hayato Ohshima, Toshiya Sato, Minesuke Yokoyama, Takashi SakuAbstract:Perlecan, a heparan sulfate proteoglycan, is enriched in the intercellular space of the enamel organ. To understand the role of Perlecan in tooth morphogenesis, we used a keratin 5 promoter to generate transgenic (Tg) mice that over-express Perlecan in epithelial cells, and examined their tooth germs at tissue and cellular levels. Immunohistochemistry showed that Perlecan was more strongly expressed in the enamel organ cells of Tg mice than in wild-type mice. Histopathology showed wider intercellular spaces in the stellate reticulum of the Tg molars and loss of cellular polarity in the enamel organ, especially in its cervical region. Hertwig's epithelial root sheath (HERS) cells in Tg mice were irregularly aligned due to excessive deposits of Perlecan along the inner, as well as on the outer sides of the HERS. Tg molars had dull-ended crowns and outward-curved tooth roots and their enamel was poorly crystallized, resulting in pronounced attrition of molar cusp areas. In Tg mice, expression of integrin β1 mRNA was remarkably higher at E18, while expression of bFGF, TGF-β1, DSPP and Shh was more elevated at P1. The overexpression of Perlecan in the enamel organ resulted in irregular morphology of teeth, suggesting that the expression of Perlecan regulates growth factor signaling in a stage-dependent manner during each step of the interaction between ameloblast-lineage cells and mesenchymal cells.
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Targeting Perlecan in Human Keratinocytes Reveals Novel Roles for Perlecan in Epidermal Formation
Journal of Biological Chemistry, 2005Co-Authors: Ifat Sher, John M. Whitelock, Simona Zisman-rozen, Liat Eliahu, Nicole Maas-szabowski, Yoshihiko Yamada, Dirk Breitkreutz, Norbert E. Fusenig, Eri Arikawa-hirasawa, Renato V. IozzoAbstract:Abstract Heparin-binding growth factors are crucial for the formation of human epidermis, but little is known about the role of heparan sulfate proteoglycans in this process. Here we investigated the role of the heparan sulfate proteoglycan, Perlecan, in the formation of human epidermis, by utilizing in vitro engineered human skin. By disrupting Perlecan expression either in the dermis or the epidermis, we found that epidermally derived Perlecan is essential for epidermal formation. Perlecan-deficient keratinocytes formed a strikingly thin and poorly organized epidermis because of premature apoptosis and failure to complete their stratification program. Exogenous Perlecan fully restored epidermal formation. Perlecan deposition in the basement membrane zone correlated with formation of multilayered epidermis. Perlecan deficiency, however, had no effect on the lining and deposition of major basement membrane components as was evident by a continuous linear staining of laminin and collagen IV. Similarly, Perlecan deficiency did not affect the distribution of β1 integrin. Addition of the Perlecan ligand, fibroblast growth factor 7, protected Perlecan-deficient keratinocytes from cell death and improved the thickness of the epidermis. Taken together, our results revealed novel roles for Perlecan in epidermal formation. Perlecan regulates both the survival and terminal differentiation steps of keratinocytes. Our results suggested a model whereby Perlecan regulates these processes via controlling the bioavailability of Perlecan-binding soluble factors involved in epidermal morphogenesis.
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Atherosclerosis in Perlecan heterozygous mice.
Journal of Lipid Research, 2004Co-Authors: Reeba K. Vikramadithyan, Yoshihiko Yamada, Eri Arikawa-hirasawa, Yuko Kako, Guangping Chen, Ira J. GoldbergAbstract:The hypothesis that lipoprotein association with Perlecan is atherogenic was tested by studying atherosclerosis in mice that had a heterozygous deletion of Perlecan, the primary extracellular heparan sulfate proteoglycan in arteries. We first studied the expression of Perlecan in mouse lesions and noted that this proteoglycan in aorta was found in the subendothelial matrix. Perlecan was also a major component of the lesional extracellular matrix. Mice with a heterozygous deletion had a reduction in arterial wall Perlecan expression. Atherosclerosis in these mice was studied after crossing the defect into the apolipoprotein E (apoE) and LDL receptor knockout backgrounds. At 12 weeks, chow-fed apoE null mice with a heterozygous deletion had less atherosclerosis. However, at 24 weeks and in the LDL receptor heterozygous background, the presence of a Perlecan knockout allele did not significantly alter lesion size. Thus, it appears that loss of Perlecan leads to less atherosclerosis in early lesions. Although this might be attributable to a decrease in lipoprotein retention, it should be noted that Perlecan might mediate multiple other processes that could, in sum, accelerate atherosclerosis.