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1.
Am J Physiol Cell Physiol ; 323(2): C505-C519, 2022 08 01.
Article in English | MEDLINE | ID: mdl-35759431

ABSTRACT

One of the main components of the extracellular matrix (ECM) of blood vessels is hyaluronic acid or hyaluronan (HA). It is a ubiquitous polysaccharide belonging to the family of glycosaminoglycans, but, differently from other proteoglycan-associated glycosaminoglycans, it is synthesized on the plasma membrane by a family of three HA synthases (HAS). HA can be released as a free polymer in the extracellular space or remain associated with the plasma membrane in the pericellular space via HAS or HA-binding proteins. Several cell surface proteins can interact with HA working as HA receptors, like CD44, RHAMM, and LYVE-1. In physiological conditions, HA is localized in the glycocalyx and the adventitia where it is responsible for the loose and hydrated vascular structure favoring flexibility and allowing the stretching of vessels in response to mechanical forces. During atherogenesis, ECM undergoes dramatic alterations that have a crucial role in lipoprotein retention and in triggering multiple signaling cascades that induce the cells to exit from their quiescent status. HA becomes highly present in the media and neointima favoring smooth muscle cells dedifferentiation, migration, and proliferation that strongly contribute to vessel wall thickening. Furthermore, HA is able to modulate immune cell recruitment both within the vessel wall and on the endothelial cell layer. This review is focused on deeply analyzing the effects of HA on vascular cell behavior.


Subject(s)
Atherosclerosis , Hyaluronic Acid , Atherosclerosis/metabolism , Endothelial Cells/metabolism , Humans , Hyaluronan Receptors/metabolism , Hyaluronic Acid/metabolism , Macrophages/metabolism , Myocytes, Smooth Muscle/metabolism
2.
Semin Cancer Biol ; 62: 9-19, 2020 05.
Article in English | MEDLINE | ID: mdl-31319162

ABSTRACT

Extracellular matrix (ECM) is a complex network of macromolecules such as proteoglycans (PGs), glycosaminoglycans (GAGs) and fibrous proteins present within all tissues and organs. The main role of ECM is not only to provide an essential mechanical scaffold for the cells but also to mediate crucial biochemical cues that are required for tissue homeostasis. Dysregulations in ECM deposition alter cell microenvironment, triggering the onset or the rapid progression of several diseases, including cancer. Hyaluronan (HA) is a ubiquitous component of ECM considered as one of the main players of cancer initiation and progression. This review discusses how HA participate in and regulate several aspects of tumorigenesis, with particular attention to the hallmarks of cancer proposed by Hanahan and Weinberg such as sustaining of the proliferative signaling, evasion of apoptosis, angiogenesis, activation of invasion and metastases, reprogramming of energy metabolism and evasion of immune response.


Subject(s)
Disease Susceptibility , Hyaluronic Acid/metabolism , Neoplasms/etiology , Neoplasms/metabolism , Animals , Cell Movement , Cell Proliferation , Cell Transformation, Neoplastic/metabolism , Energy Metabolism , Extracellular Matrix/metabolism , Humans , Neoplasm Invasiveness , Neoplasm Metastasis , Neoplasms/pathology , Neovascularization, Pathologic/metabolism , Signal Transduction , Tumor Escape , Tumor Microenvironment
3.
J Biol Chem ; 295(11): 3485-3496, 2020 03 13.
Article in English | MEDLINE | ID: mdl-31932306

ABSTRACT

Hyaluronan (HA) is one of the most prevalent glycosaminoglycans of the vascular extracellular matrix (ECM). Abnormal HA accumulation within blood vessel walls is associated with tissue inflammation and is prominent in most vascular pathological conditions such as atherosclerosis and restenosis. Hyaluronan synthase 2 (HAS2) is the main hyaluronan synthase enzyme involved in HA synthesis and uses cytosolic UDP-glucuronic acid and UDP-GlcNAc as substrates. The synthesis of UDP-glucuronic acid can alter the NAD+/NADH ratio via the enzyme UDP-glucose dehydrogenase, which oxidizes the alcohol group at C6 to the COO- group. Here, we show that HAS2 expression can be modulated by sirtuin 1 (SIRT1), the master metabolic sensor of the cell, belonging to the class of NAD+-dependent deacetylases. Our results revealed the following. 1) Treatments of human aortic smooth muscle cells (AoSMCs) with SIRT1 activators (SRT1720 and resveratrol) inhibit both HAS2 expression and accumulation of pericellular HA coats. 2) Tumor necrosis factor α (TNFα) induced HA-mediated monocyte adhesion and AoSMC migration, whereas SIRT1 activation prevented immune cell recruitment and cell motility by reducing the expression levels of the receptor for HA-mediated motility, RHAMM, and the HA-binding protein TNF-stimulated gene 6 protein (TSG6). 3) SIRT1 activation prevented nuclear translocation of NF-κB (p65), which, in turn, reduced the levels of HAS2-AS1, a long-noncoding RNA that epigenetically controls HAS2 mRNA expression. In conclusion, we demonstrate that both HAS2 expression and HA accumulation by AoSMCs are down-regulated by the metabolic sensor SIRT1.


Subject(s)
Cell Nucleus/metabolism , Gene Expression Regulation , Hyaluronan Synthases/genetics , NF-kappa B/metabolism , RNA, Long Noncoding/genetics , Sirtuin 1/metabolism , Aorta/cytology , Cell Nucleus/drug effects , Cells, Cultured , Cytoprotection/drug effects , Extracellular Matrix/metabolism , Gene Expression Regulation/drug effects , Heterocyclic Compounds, 4 or More Rings/pharmacology , Humans , Hyaluronan Synthases/metabolism , Hyaluronic Acid/metabolism , Inflammation/pathology , Models, Biological , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Protein Transport/drug effects , Resveratrol/pharmacology , Tumor Necrosis Factor-alpha
4.
Int J Mol Sci ; 22(15)2021 Jul 28.
Article in English | MEDLINE | ID: mdl-34360868

ABSTRACT

Cancer is a multifaceted and complex pathology characterized by uncontrolled cell proliferation and decreased apoptosis. Most cancers are recognized by an inflammatory environment rich in a myriad of factors produced by immune infiltrate cells that induce host cells to differentiate and to produce a matrix that is more favorable to tumor cells' survival and metastasis. As a result, the extracellular matrix (ECM) is changed in terms of macromolecules content, degrading enzymes, and proteins. Altered ECM components, derived from remodeling processes, interact with a variety of surface receptors triggering intracellular signaling that, in turn, cancer cells exploit to their own benefit. This review aims to present the role of different aspects of ECM components in the tumor microenvironment. Particularly, we highlight the effect of pro- and inflammatory factors on ECM degrading enzymes, such as metalloproteases, and in a more detailed manner on hyaluronan metabolism and the signaling pathways triggered by the binding of hyaluronan with its receptors. In addition, we sought to explore the role of extracellular chaperones, especially of clusterin which is one of the most prominent in the extracellular space, in proteostasis and signaling transduction in the tumor microenvironment. Although the described tumor microenvironment components have different biological roles, they may engage common signaling pathways that favor tumor growth and metastasis.


Subject(s)
Extracellular Matrix/metabolism , Inflammation , Neoplasms , Proteostasis , Tumor Microenvironment , Humans , Inflammation/metabolism , Inflammation/pathology , Neoplasms/metabolism , Neoplasms/pathology
5.
Chem Rev ; 118(18): 9152-9232, 2018 09 26.
Article in English | MEDLINE | ID: mdl-30204432

ABSTRACT

The extracellular matrix (ECM) constitutes a highly dynamic three-dimensional structural network comprised of macromolecules, such as proteoglycans/glycosaminoglycans (PGs/GAGs), collagens, laminins, fibronectin, elastin, other glycoproteins and proteinases. In recent years, the field of PGs has expanded rapidly. Due to their high structural complexity and heterogeneity, PGs mediate several homeostatic and pathological processes. PGs consist of a protein core and one or more covalently attached GAG chains, which provide the protein cores with the ability to interact with several proteins. The GAG building blocks of PGs significantly influence the chemical and functional properties of PGs. The primary goal of this comprehensive review is to summarize major achievements and paradigm-shifting discoveries made on the PG/GAG chemistry-biology axis, focusing on structural variability, structure-function relationships, metabolic, molecular, and epigenetic mechanisms underlying their synthesis. Recent insights related to exosome biogenesis, degradation, and cell signaling, their status as diagnostic tools and potential pharmacological targets in diseases as well as current applications in nanotechnology and biotechnology are addressed. Moreover, issues related to docking studies, molecular modeling, GAG/PG interaction networks, and their integration are discussed.


Subject(s)
Glycosaminoglycans/chemistry , Glycosaminoglycans/physiology , Proteoglycans/chemistry , Proteoglycans/physiology , Animals , Cell Line, Tumor , Epigenesis, Genetic , Extracellular Matrix/metabolism , Glycosaminoglycans/genetics , Humans , Neoplasms/physiopathology , Neurodegenerative Diseases/physiopathology , Protein Domains , Proteoglycans/genetics , Signal Transduction/physiology
6.
Adv Exp Med Biol ; 1245: 147-161, 2020.
Article in English | MEDLINE | ID: mdl-32266657

ABSTRACT

The biology of tumor cells strictly depends on their microenvironment architecture and composition, which controls the availability of growth factors and signaling molecules. Thus, the network of glycosaminoglycans, proteoglycans, and proteins known as extracellular matrix (ECM) that surrounds the cells plays a central role in the regulation of tumor fate. Heparan sulfate (HS) and heparan sulfate proteoglycans (HSPGs) are highly versatile ECM components that bind and regulate the activity of growth factors, cell membrane receptors, and other ECM molecules. These HS binding partners modulate cell adhesion, motility, and proliferation that are processes altered during tumor progression. Modification in the expression and activity of HS, HSPGs, and the respective metabolic enzymes results unavoidably in alteration of tumor cell microenvironment. In this light, the targeting of HS structure and metabolism is potentially a new tool in the treatment of different cancer types.


Subject(s)
Heparitin Sulfate , Neoplasms , Tumor Microenvironment , Extracellular Matrix/metabolism , Heparan Sulfate Proteoglycans/metabolism , Heparitin Sulfate/metabolism , Humans , Neoplasms/metabolism , Neoplasms/pathology
7.
J Biol Chem ; 293(43): 16951-16952, 2018 10 26.
Article in English | MEDLINE | ID: mdl-30366972

ABSTRACT

Nutrient sensing is a critical cell function that regulates survival and growth by adjusting metabolism. During nutrient shortage, autophagy enables the recycling of major cellular components to prevent cell death. Understanding the mechanisms that trigger and control autophagy is of fundamental importance, as this degradative pathway plays a pivotal role in many diseases. Gubbiotti et al. report the identification of a new player, the proteoglycan decorin, which functions as a nutrient sensor in the extracellular matrix and controls autophagy in the heart.


Subject(s)
Autophagy , Decorin/physiology , Extracellular Matrix/metabolism , Metabolome , Myocytes, Cardiac/pathology , Nutrients/metabolism , Animals , Cellular Reprogramming , Humans , Myocytes, Cardiac/metabolism
8.
J Biol Chem ; 293(52): 20214-20226, 2018 12 28.
Article in English | MEDLINE | ID: mdl-30377255

ABSTRACT

Conophylline is a Vinca alkaloid from leaves of the tropical plant Ervatamia microphylla and has been shown to mimic the effect of the growth and differentiation factor activin A on pancreatic progenitor cells. However, activin A stimulates fibrosis of pancreatic stellate cells, whereas conophylline inhibits it, suggesting that this compound may serve as an antifibrotic drug. Here we investigated the effects of conophylline on human foreskin fibroblasts, especially focusing on extracellular matrix (ECM) proteins. A gene microarray analysis revealed that conophylline remarkably suppressed expression of the gene for hyaluronan synthase 2 (HAS2) and of its antisense RNA, whereas the expression of collagen genes was unaffected. Of note, immunostaining experiments revealed that conophylline substantially inhibits incorporation of versican and collagens into the ECM in cells treated with transforming growth factor ß (TGFß), which promotes collagen synthesis, but not in cells not treated with TGFß. Moreover, a protein biosynthesis assay disclosed that conophylline decreases collagen biosynthesis, concomitant with a decrease in total protein biosynthesis, indicating that conophylline-mediated inhibition of fibrosis is not specific to collagen synthesis. Conophylline affected neither TGFß-induced nuclear translocation of SMAD family member 2/3 (SMAD2/3) nor phosphorylation of SMAD2. However, conophylline substantially inhibited phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), suggesting that conophylline inhibits HAS2 expression via TGFß-mediated activation of the ERK1/2 pathway. Taken together, our results indicate that conophylline may be a useful inhibitor of ECM formation in fibrosis.


Subject(s)
Extracellular Matrix/metabolism , MAP Kinase Signaling System/drug effects , Vinca Alkaloids/pharmacology , Cells, Cultured , Collagen/metabolism , Fibroblasts , Gene Expression Regulation, Enzymologic/drug effects , Humans , Hyaluronan Synthases/biosynthesis , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/metabolism , Phosphorylation/drug effects , Protein Biosynthesis/drug effects , Smad2 Protein/metabolism , Smad3 Protein/metabolism , Versicans/metabolism
9.
Glycoconj J ; 34(3): 411-420, 2017 06.
Article in English | MEDLINE | ID: mdl-27744520

ABSTRACT

Proteoglycans and glycosaminoglycans modulate numerous cellular processes relevant to tumour progression, including cell proliferation, cell-matrix interactions, cell motility and invasive growth. Among the glycosaminoglycans with a well-documented role in tumour progression are heparan sulphate, chondroitin/dermatan sulphate and hyaluronic acid/hyaluronan. While the mode of biosynthesis differs for sulphated glycosaminoglycans, which are synthesised in the ER and Golgi compartments, and hyaluronan, which is synthesized at the plasma membrane, these polysaccharides partially compete for common substrates. In this study, we employed a siRNA knockdown approach for heparan sulphate (EXT1) and heparan/chondroitin/dermatan sulphate-biosynthetic enzymes (ß4GalT7) in the aggressive human breast cancer cell line MDA-MB-231 to study the impact on cell behaviour and hyaluronan biosynthesis. Knockdown of ß4GalT7 expression resulted in a decrease in cell viability, motility and adhesion to fibronectin, while these parameters were unchanged in EXT1-silenced cells. Importantly, these changes were associated with a decreased expression of syndecan-1, decreased signalling response to HGF and an increase in the synthesis of hyaluronan, due to an upregulation of the hyaluronan synthases HAS2 and HAS3. Interestingly, EXT1-depleted cells showed a downregulation of the UDP-sugar transporter SLC35D1, whereas SLC35D2 was downregulated in ß4GalT7-depleted cells, indicating an intricate regulatory network that connects all glycosaminoglycans synthesis. The results of our in vitro study suggest that a modulation of breast cancer cell behaviour via interference with heparan sulphate biosynthesis may result in a compensatory upregulation of hyaluronan biosynthesis. These findings have important implications for the development of glycosaminoglycan-targeted therapeutic approaches for malignant diseases.


Subject(s)
Chondroitin Sulfates/biosynthesis , Dermatan Sulfate/analogs & derivatives , Epithelial Cells/metabolism , Gene Expression Regulation, Neoplastic , Heparitin Sulfate/biosynthesis , Hyaluronic Acid/biosynthesis , Cell Adhesion , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cell Survival , Chondroitin Sulfates/antagonists & inhibitors , Chondroitin Sulfates/genetics , Dermatan Sulfate/antagonists & inhibitors , Dermatan Sulfate/biosynthesis , Dermatan Sulfate/genetics , Epithelial Cells/pathology , Extracellular Matrix/chemistry , Extracellular Matrix/metabolism , Heparitin Sulfate/antagonists & inhibitors , Heparitin Sulfate/genetics , Humans , Hyaluronan Synthases/antagonists & inhibitors , Hyaluronan Synthases/genetics , Hyaluronan Synthases/metabolism , Hyaluronic Acid/antagonists & inhibitors , Hyaluronic Acid/genetics , Isoenzymes/antagonists & inhibitors , Isoenzymes/genetics , Isoenzymes/metabolism , Mammary Glands, Human/metabolism , Mammary Glands, Human/pathology , Monosaccharide Transport Proteins/antagonists & inhibitors , Monosaccharide Transport Proteins/genetics , Monosaccharide Transport Proteins/metabolism , N-Acetylglucosaminyltransferases/antagonists & inhibitors , N-Acetylglucosaminyltransferases/genetics , N-Acetylglucosaminyltransferases/metabolism , N-Acetyllactosamine Synthase/antagonists & inhibitors , N-Acetyllactosamine Synthase/genetics , N-Acetyllactosamine Synthase/metabolism , Nucleotide Transport Proteins/antagonists & inhibitors , Nucleotide Transport Proteins/genetics , Nucleotide Transport Proteins/metabolism , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Signal Transduction
10.
J Biol Chem ; 289(42): 28816-26, 2014 Oct 17.
Article in English | MEDLINE | ID: mdl-25183006

ABSTRACT

Changes in the microenvironment organization within vascular walls are critical events in the pathogenesis of vascular pathologies, including atherosclerosis and restenosis. Hyaluronan (HA) accumulation into artery walls supports vessel thickening and is involved in many cardiocirculatory diseases. Excessive cytosolic glucose can enter the hexosamine biosynthetic pathway, increase UDP-N-acetylglucosamine (UDP-GlcNAc) availability, and lead to modification of cytosolic proteins via O-linked attachment of the monosaccharide ß-N-GlcNAc (O-GlcNAcylation) from UDP-GlcNAc by the enzyme O-GlcNAc transferase. As many cytoplasmic and nuclear proteins can be glycosylated by O-GlcNAc, we studied whether the expression of the HA synthases that synthesize HA could be controlled by O-GlcNAcylation in human aortic smooth muscle cells. Among the three HAS isoenzymes, only HAS2 mRNA increased after O-GlcNAcylation induced by glucosamine treatments or by inhibiting O-GlcNAc transferase with PUGNAC (O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate). We found that the natural antisense transcript of HAS2 (HAS2-AS1) was absolutely necessary to induce the transcription of the HAS2 gene. Moreover, we found that O-GlcNAcylation modulated HAS2-AS1 promoter activation by recruiting the NF-κB subunit p65, but not the HAS2 promoter, whereas HAS2-AS1 natural antisense transcript, working in cis, regulated HAS2 transcription by altering the chromatin structure around the HAS2 proximal promoter via O-GlcNAcylation and acetylation. These results indicate that HAS2 transcription can be finely regulated not only by recruiting transcription factors to the promoter as previously described but also by modulating chromatin accessibility by epigenetic modifications.


Subject(s)
Gene Expression Regulation, Enzymologic , Glucuronosyltransferase/genetics , Acetylglucosamine/chemistry , Animals , Aorta/enzymology , Base Sequence , Cell Nucleus/enzymology , Chromatin/chemistry , Cytoplasm/enzymology , Epigenesis, Genetic , Gene Silencing , Glucuronosyltransferase/physiology , Humans , Hyaluronan Synthases , Male , Mice , Mice, Knockout , Models, Genetic , Molecular Sequence Data , Monosaccharides/chemistry , Myocytes, Smooth Muscle/enzymology , N-Acetylglucosaminyltransferases/chemistry , Promoter Regions, Genetic , RNA, Messenger/metabolism , Transcription, Genetic
11.
Biochim Biophys Acta ; 1840(8): 2452-9, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24513306

ABSTRACT

BACKGROUND: Hyaluronan is a critical component of extracellular matrix with several different roles. Besides the contribution to the tissue hydration, mechanical properties and correct architecture, hyaluronan plays important biological functions interacting with different molecules and receptors. SCOPE OF REVIEW: The review addresses the control of hyaluronan synthesis highlighting the critical role of hyaluronan synthase 2 in this context as well as discussing the recent findings related to covalent modifications which influence the enzyme activity. Moreover, the interactions with specific receptors and hyaluronan are described focusing on the importance of polymer size in the modulation of hyaluronan signaling. MAJOR CONCLUSIONS: Due to its biological effects on cells recently described, it is evident how hyaluronan is to be considered not only a passive component of extracellular matrix but also an actor involved in several scenarios of cell behavior. GENERAL SIGNIFICANCE: The effects of metabolism on the control of hyaluronan synthesis both in healthy and pathologic conditions are critical and still not completely understood. The hyaluronan capacity to bind several receptors triggering specific pathways may represent a valid target for new approach in several therapeutic strategies. This article is part of a Special Issue entitled Matrix-mediated cell behaviour and properties.


Subject(s)
Hyaluronic Acid/biosynthesis , Signal Transduction , Animals , Biosynthetic Pathways , Humans , Hyaluronan Receptors/chemistry , Hyaluronan Receptors/metabolism , Hyaluronic Acid/chemistry
12.
Glycoconj J ; 32(3-4): 93-103, 2015 May.
Article in English | MEDLINE | ID: mdl-25971701

ABSTRACT

The hyaluronan (HA) polymer is a critical component of extracellular matrix with a remarkable structure: is a linear and unbranched polymer without sulphate or phosphate groups. It is ubiquitous in mammals showing several biological functions, ranging from cell proliferation and migration to angiogenesis and inflammation. For its critical biological functions the amount of HA in tissues is carefully controlled by different mechanisms including covalent modification of the synthetic enzymes and epigenetic control of their gene expression. The concentration of HA is also critical in several pathologies including cancer, diabetes and inflammation. Beside these biological roles, the structural properties of HA allow it to take advantage of its capacity to form gels even at concentration of 1 % producing scaffolds with very promising applications in regenerative medicine as biocompatible material for advanced therapeutic uses. In this review we highlight the biological aspects of HA addressing the mechanisms controlling the HA content in tissues as well as its role in important human pathologies. In the second part of the review we highlight the different use of HA polymers in the modern biotechnology.


Subject(s)
Biotechnology/methods , Hyaluronic Acid/chemistry , Hyaluronic Acid/metabolism , Animals , Dietary Supplements , Drug Delivery Systems , Extracellular Matrix/metabolism , Humans , Hyaluronic Acid/administration & dosage , Inflammation/metabolism , Neoplasms/metabolism
13.
J Biol Chem ; 288(41): 29595-603, 2013 Oct 11.
Article in English | MEDLINE | ID: mdl-23979132

ABSTRACT

Thickening of the vessel in response to high low density lipoprotein(s) (LDL) levels is a hallmark of atherosclerosis, characterized by increased hyaluronan (HA) deposition in the neointima. Human native LDL trapped within the arterial wall undergoes modifications such as oxidation (oxLDL). The aim of our study is to elucidate the link between internalization of oxLDL and HA production in vitro, using human aortic smooth muscle cells. LDL were used at an effective protein concentration of 20-50 µg/ml, which allowed 80% cell viability. HA content in the medium of untreated cells was 28.9 ± 3.7 nmol HA-disaccharide/cell and increased after oxLDL treatment to 53.9 ± 5.6. OxLDL treatments doubled the transcripts of HA synthase HAS2 and HAS3. Accumulated HA stimulated migration of aortic smooth muscle cells and monocyte adhesiveness to extracellular matrix. The effects induced by oxLDL were inhibited by blocking LOX-1 scavenger receptor with a specific antibody (10 µg/ml). The cholesterol moiety of LDL has an important role in HA accumulation because cholesterol-free oxLDL failed to induce HA synthesis. Nevertheless, cholesterol-free oxLDL and unmodified cholesterol (20 µg/ml) induce only HAS3 transcription, whereas 22,oxysterol affects both HAS2 and HAS3. Moreover, HA deposition was associated with higher expression of endoplasmic reticulum stress markers (CHOP and GRP78). Our data suggest that HA synthesis can be induced in response to specific oxidized sterol-related species delivered through oxLDL.


Subject(s)
Cell Movement/drug effects , Hyaluronic Acid/biosynthesis , Lipoproteins, LDL/pharmacology , Myocytes, Smooth Muscle/drug effects , Antibodies/immunology , Antibodies/pharmacology , Aorta/cytology , Cell Adhesion/drug effects , Cell Survival/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Endoplasmic Reticulum Chaperone BiP , Extracellular Matrix/metabolism , Gene Expression/drug effects , Glucuronosyltransferase/genetics , Humans , Hyaluronan Synthases , Lipoproteins, LDL/metabolism , Monocytes/drug effects , Monocytes/metabolism , Myocytes, Smooth Muscle/metabolism , Oxidation-Reduction , Reverse Transcriptase Polymerase Chain Reaction , Scavenger Receptors, Class E/antagonists & inhibitors , Scavenger Receptors, Class E/immunology , Scavenger Receptors, Class E/metabolism , U937 Cells
14.
Proc Natl Acad Sci U S A ; 108(3): 1104-9, 2011 Jan 18.
Article in English | MEDLINE | ID: mdl-21189302

ABSTRACT

A recent body of evidence indicates an active role for stromal (mis)-regulation in the progression of neoplasias. Within this conceptual framework, genes belonging to the growing but still poorly characterized class of tumor antagonizing/malignancy suppressor genes (TAG/MSG) seem to play a crucial role in the regulation of the cross-talk between stromal and epithelial cells by controlling malignant growth in vivo without affecting any cancer-related phenotype in vitro. Here, we have functionally characterized the human RNASET2 gene, which encodes the first human member of the widespread Rh/T2/S family of extracellular RNases and was recently found to be down-regulated at the transcript level in several primary ovarian tumors or cell lines and in melanoma cell lines. Although we could not detect any activity for RNASET2 in several functional in vitro assays, a remarkable control of ovarian tumorigenesis could be detected in vivo. Moreover, the control of ovarian tumorigenesis mediated by this unique tumor suppressor gene occurs through modification of the cellular microenvironment and the induction of immunocompetent cells of the monocyte/macrophage lineage. Taken together, the data presented in this work strongly indicate RNASET2 as a previously unexplored member of the growing family of tumor-antagonizing genes.


Subject(s)
Macrophages/immunology , Ovarian Neoplasms/genetics , Ribonucleases/immunology , Tumor Suppressor Proteins/immunology , Analysis of Variance , Animals , Cell Line, Tumor , Female , Humans , Immunohistochemistry , In Situ Hybridization , In Vitro Techniques , Mice , Mice, Nude , Ovarian Neoplasms/pathology , Ribonucleases/genetics , Tumor Suppressor Proteins/genetics , Xenograft Model Antitumor Assays
15.
J Biol Chem ; 287(42): 35544-35555, 2012 Oct 12.
Article in English | MEDLINE | ID: mdl-22887999

ABSTRACT

Hyaluronan (HA) is a glycosaminoglycan present in most tissue microenvironments that can modulate many cell behaviors, including proliferation, migration, and adhesive proprieties. In contrast with other glycosaminoglycans, which are synthesized in the Golgi, HA is synthesized at the plasma membrane by one or more of the three HA synthases (HAS1-3), which use cytoplasmic UDP-glucuronic acid and UDP-N-acetylglucosamine as substrates. Previous studies revealed the importance of UDP-sugars for regulating HA synthesis. Therefore, we analyzed the effect of UDP-GlcNAc availability and protein glycosylation with O-linked N-acetylglucosamine (O-GlcNAcylation) on HA and chondroitin sulfate synthesis in primary human aortic smooth muscle cells. Glucosamine treatment, which increases UDP-GlcNAc availability and protein O-GlcNAcylation, increased synthesis of both HA and chondroitin sulfate. However, increasing O-GlcNAcylation by stimulation with O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate without a concomitant increase of UDP-GlcNAc increased only HA synthesis. We found that HAS2, the main synthase in aortic smooth muscle cells, can be O-GlcNAcylated on serine 221, which strongly increased its activity and its stability (t(½) >5 h versus ∼17 min without O-GlcNAcylation). S221A mutation prevented HAS2 O-GlcNAcylation, which maintained the rapid turnover rate even in the presence of GlcN and increased UDP-GlcNAc. These findings could explain the elevated matrix HA observed in diabetic vessels that, in turn, could mediate cell dedifferentiation processes critical in vascular pathologies.


Subject(s)
Aorta/metabolism , Chondroitin Sulfates/biosynthesis , Diabetic Angiopathies/metabolism , Glucuronosyltransferase/metabolism , Hyaluronic Acid/biosynthesis , Muscle Proteins/metabolism , Muscle, Smooth, Vascular/metabolism , Uridine Diphosphate N-Acetylglucosamine/metabolism , Amino Acid Substitution , Aorta/pathology , Cell Line , Chondroitin Sulfates/genetics , Diabetic Angiopathies/genetics , Diabetic Angiopathies/pathology , Glucuronosyltransferase/genetics , Glycosylation , Humans , Hyaluronan Synthases , Hyaluronic Acid/genetics , Muscle Proteins/genetics , Muscle, Smooth, Vascular/pathology , Mutation, Missense , Uridine Diphosphate N-Acetylglucosamine/genetics
16.
Cancers (Basel) ; 15(3)2023 Jan 28.
Article in English | MEDLINE | ID: mdl-36765756

ABSTRACT

The presence of the glycosaminoglycan hyaluronan in the extracellular matrix of tissues is the result of the cooperative synthesis of several resident cells, that is, macrophages and tumor and stromal cells. Any change in hyaluronan concentration or dimension leads to a modification in stiffness and cellular response through receptors on the plasma membrane. Hyaluronan has an effect on all cancer cell behaviors, such as evasion of apoptosis, limitless replicative potential, sustained angiogenesis, and metastasis. It is noteworthy that hyaluronan metabolism can be dramatically altered by growth factors and matrikines during inflammation, as well as by the metabolic homeostasis of cells. The regulation of HA deposition and its dimensions are pivotal for tumor progression and cancer patient prognosis. Nevertheless, because of all the factors involved, modulating hyaluronan metabolism could be tough. Several commercial drugs have already been described as potential or effective modulators; however, deeper investigations are needed to study their possible side effects. Moreover, other matrix molecules could be identified and targeted as upstream regulators of synthetic or degrading enzymes. Finally, co-cultures of cancer, fibroblasts, and immune cells could reveal potential new targets among secreted factors.

17.
Cancers (Basel) ; 15(15)2023 Jul 27.
Article in English | MEDLINE | ID: mdl-37568628

ABSTRACT

The expression of the estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2) in breast cancer cells is critical for determining tumor aggressiveness and targeting therapies. The presence of such receptors allows for the use of antagonists that effectively reduce breast cancer growth and dissemination. However, the absence of such receptors in triple-negative breast cancer (TNBC) reduces the possibility of targeted therapy, making these tumors very aggressive with a poor outcome. Cancers are not solely composed of tumor cells, but also include several types of infiltrating cells, such as fibroblasts, macrophages, and other immune cells that have critical functions in regulating cancer cell behaviors. In addition to these cells, the extracellular matrix (ECM) has become an important player in many aspects of breast cancer biology, including cell growth, motility, metabolism, and chemoresistance. Hyaluronan (HA) is a key ECM component that promotes cell proliferation and migration in several malignancies. Notably, HA accumulation in the tumor stroma is a negative prognostic factor in breast cancer. HA metabolism depends on the fine balance between HA synthesis by HA synthases and degradation yielded by hyaluronidases. All the different cell types present in the tumor can release HA in the ECM, and in this review, we will describe the role of HA and HA metabolism in different breast cancer subtypes.

18.
Methods Mol Biol ; 2619: 53-60, 2023.
Article in English | MEDLINE | ID: mdl-36662461

ABSTRACT

Hyaluronan (HA) is the most abundant glycosaminoglycan in the extracellular matrix, and its deposition is strictly related to changes in cellular behaviors, such as cell migration, proliferation, and adhesion. Pericellular HA is abundant in a variety of cell types, and its amount could reflect specific conditions, thus suggesting a particular cellular status.Particle exclusion assay is a useful tool to visualize pericellular matrices with a high HA content, simply employing microscope image analysis. This approach is quick and allows to visualize the presence of a clear pericellular region around single cells, where fixed red blood cells are excluded if the pericellular matrix has been deposited.


Subject(s)
Extracellular Matrix , Hyaluronic Acid , Hyaluronic Acid/metabolism , Extracellular Matrix/metabolism , Cell Movement , Hyaluronan Receptors/metabolism , Hyaluronan Synthases/metabolism
19.
Biomolecules ; 14(1)2023 Dec 24.
Article in English | MEDLINE | ID: mdl-38254629

ABSTRACT

Vascular calcification (VC) is an age-related complication characterised by calcium-phosphate deposition in the arterial wall driven by the osteogenic transformation of vascular smooth muscle cells (VSMCs). The JAK-STAT pathway is an emerging target in inflammation. Considering the relationship between VC and inflammation, we investigated the role of JAK-STAT signalling during VSMC calcification. Human aortic smooth muscle cells (HASMCs) were cultured in high-inorganic phosphate (Pi) medium for up to 7 days; calcium deposition was determined via Alizarin staining and colorimetric assay. Inflammatory factor secretion was evaluated via ELISA and JAK-STAT members' activation using Western blot or immunohistochemistry on HASMCs or calcified aortas of Vitamin D-treated C57BL6/J mice, respectively. The JAK-STAT pathway was blocked by JAK Inhibitor I and Von Kossa staining was used for calcium deposits in murine aortic rings. During Pi-induced calcification, HASMCs released IL-6, IL-8, and MCP-1 and activated JAK1-JAK3 proteins and STAT1. Phospho-STAT1 was detected in murine calcified aortas. Blocking of the JAK-STAT cascade reduced HASMC proliferation and pro-inflammatory factor expression and release while increasing calcium deposition and osteogenic transcription factor RUNX2 expression. Consistently, JAK-STAT pathway inhibition exacerbates mouse aortic ring calcification ex vivo. Intriguingly, our results suggest an alternative link between VSMC inflammation and VC.


Subject(s)
Muscle, Smooth, Vascular , Vascular Calcification , Humans , Animals , Mice , Calcium , Janus Kinases , STAT Transcription Factors , Signal Transduction , Vascular Calcification/chemically induced , Inflammation
20.
J Biol Chem ; 286(10): 7917-7924, 2011 Mar 11.
Article in English | MEDLINE | ID: mdl-21228273

ABSTRACT

Hyaluronan (HA) is an extracellular matrix glycosaminoglycan (GAG) involved in cell motility, proliferation, tissue remodeling, development, differentiation, inflammation, tumor progression, and invasion and controls vessel thickening in cardiovascular diseases. Therefore, the control of HA synthesis could permit the fine-tuning of cell behavior, but the mechanisms that regulate HA synthesis are largely unknown. Recent studies suggest that the availability of the nucleotide-sugar precursors has a critical role. Because the formation of UDP-sugars is a highly energetically demanding process, we have analyzed whether the energy status of the cell could control GAG production. AMP-activated protein kinase (AMPK) is the main ATP/AMP sensor of mammalian cells, and we mimicked an energy stress by treating human aortic smooth muscle cells (AoSMCs) with the AMPK activators 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside and metformin. Under these conditions, HA synthesis, but not that of the other GAGs, was greatly reduced. We confirmed the inhibitory effect of AMPK using a specific inhibitor and knock-out cell lines. We found that AMPK phosphorylated Thr-110 of human HAS2, which inhibits its enzymatic activity. In contrast, the other two HAS isoenzymes (HAS1 and HAS3) were not modified by the kinase. The reduction of HA decreased the ability of AoSMCs to proliferate, migrate, and recruit immune cells, thereby reducing the pro-atherosclerotic AoSMC phenotype. Interestingly, such effects were not recovered by treatment with exogenous HA, suggesting that AMPK can block the pro-atherosclerotic signals driven by HA by interaction with its receptors.


Subject(s)
Aorta/metabolism , Glucuronosyltransferase/metabolism , Hyaluronic Acid/biosynthesis , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , AMP-Activated Protein Kinases , Aorta/cytology , Cell Line , Cell Movement/physiology , Cell Proliferation , Gene Knockdown Techniques , Glucuronosyltransferase/genetics , Humans , Hyaluronan Synthases , Hyaluronic Acid/genetics , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytology , Signal Transduction/physiology , Stress, Physiological/physiology
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