Héparanes sulfate : Structure, fonctions, régulation

Les héparanes sulfate (HS) sont des polysaccharides complexes appartenant à la famille des glycosaminoglycanes (GAGs), présents en abondance à la surface cellulaire et dans les matrices interstitielles. De par leur positionnement stratégique à l'interface entre la cellule et son micro-environnement et leur capacité à fixer et moduler l'activité d'un très grand nombre de protéines, les HS sont impliqués dans de nombreux processus physiologiques et pathologiques. Les travaux présentés viseront à illustrer l'importance biologique de ces polysaccharides et de leurs interactions avec les protéines, principalement dans les domaines des relations hôtes-pathogènes et des processus inflammatoires. Ils s'attacheront également à montrer les difficultés que présente l'étude de ces molécules, ainsi que les méthodologies développées afin de faciliter leur caractérisation structurale et fonctionnelle. Enfin, des perspectives de recherche seront proposées, visant à intégrer les études en cours dans un contexte physiologique, par l'analyse des HS exprimés par les cellules. La structure des HS, et donc leurs propriétés biologiques, varie en effet de manière considérable selon le type cellulaire étudié, son niveau d'activation ou de différenciation. La réponse cellulaire à des stimuli externes (protéines de signalisation, cytokines pro-inflammatoires, molécules intervenant dans la reconnaissance hôte/pathogènes...) est donc intimement liée au " paysage glycanique" présent à sa surface et à sa capacité à moduler ce paysage. La compréhension de ces mécanismes représente donc un enjeu évident pour l'étude de nombreuses fonctions cellulaires

A microscale double labelling of GAG oligosaccharides compatible with enzymatic treatment and mass spectrometry

International audienceA novel double labelling of glycosaminoglycans (GAG) oligosaccharides by thia-Michael addition and deuterium incorporation at the non-reducing and reducing ends, respectively, was introduced. This was demonstrated to be both compatible with the heparin microgram scale and amenable for mass spectrometry analysis, without impairing enzymatic activities such as heparinase I and sulfatase HSulf-2

Post-Synthetic Regulation of HS Structure: The Yin and Yang of the Sulfs in Cancer.

International audienceHeparan sulfate (HS) is a complex polysaccharide that takes part in most major cellular processes, through its ability to bind and modulate a very large array of proteins. These interactions involve saccharide domains of specific sulfation pattern (S-domains), the assembly of which is tightly orchestrated by a highly regulated biosynthesis machinery. Another level of structural control does also take place at the cell surface, where degrading enzymes further modify HS post-synthetically. Amongst them are the Sulfs, a family of extracellular sulfatases (two isoforms in human) that catalyze the specific 6-O-desulfation of HS. By targeting HS functional sulfated domains, Sulfs dramatically alter its ligand binding properties, thereby modulating a broad range of signaling pathways. Consequently, Sulfs play major roles during development, as well as in tissue homeostasis and repair. Sulfs have also been associated with many pathologies including cancer, but despite increasing interest, the role of Sulfs in tumor development still remains unclear. Studies have been hindered by a poor understanding of the Sulf enzymatic activities and conflicting data have shown either anti-oncogenic or tumor-promoting effects of these enzymes, depending on the tumor models analyzed. These opposite effects clearly illustrate the fine tuning of HS functions by the Sulfs, and the need to clarify the mechanisms involved. In this review, we will detail the present knowledge on the structural and functional properties of the Sulfs, with a special focus on their implication during tumor progression. Finally, we will discuss attempts and perspectives of using the Sulfs as a biomarker of cancer prognosis and diagnostic and as a target for anti-cancer therapies

Heparan sulphate proteoglycans and viral vectors : ally or foe?

International audienceThe attachment of viruses to the host cell surface is a critical stage that will largely condition cell permissivity and productive infection. The understanding of such mechanisms is therefore essential for gene therapy applications involving viruses, as this step will influence both targeting and delivery efficiency of the gene of interest. Viral attachment depends upon the recognition and binding of viral envelope/capsid proteins to specific cellular receptors that can be from very diverse origins. Amongst them are heparan sulphate proteoglycans (HSPGs), a family of glycoproteins which, through the large binding properties of their heparan sulphate (HS) polysaccharide chains, serve as attachment receptor for a great number of viruses. The aim of this review is to provide an update on the multiple roles of HSPGs during viral infection, with a special focus on viruses used as gene delivery vectors. Consequences of HS binding for gene therapy applications will be assessed, as well as the various strategies that have been developed to potentiate the advantages or to overcome the drawbacks resulting from viral vector interaction with HS

Endocan in cancers: a lesson from a circulating dermatan sulfate proteoglycan.

International audienceAs most proteoglycans exert their biological activities in the pericellular region, circulating Endocan has appeared since its discovery as an atypical dermatan sulfate proteoglycan, with distinctive structural and functional properties. Endocan is naturally expressed by endothelial cells, highly regulated in presence of proinflammatory and proangiogenic molecules, binds to matrix proteins, growth factors, integrin, and cells, and may be then considered as an accurate marker of endothelial activation. Consequently, Endocan expression has been associated with a growing number of pathological conditions where endothelium gets challenged and notably in highly vascularized cancers. In this context, Endocan has indeed been rapidly emerging as a promising tissue- and blood-based marker of the vascular growth and neoangiogenesis during cancer progression. Furthermore, very recent studies have reported an expression of Endocan by the tumor cells themselves. This highlights Endocan as a multifaceted molecule with a great interest for researchers and clinicians to better understand tumor development, from the bench to the clinics. With promising perspectives of clinical applications, Endocan thus appears as an exciting model for on going and future developments of proteoglycan-based approaches in cancer diagnostics and/or therapy

Sweet but Challenging: Tackling the Complexity of GAGs with Engineered Tailor‐Made Biomaterials

Glycosaminoglycans (GAGs) play a crucial role in tissue homeostasis by regulating the activity and diffusion of bioactive molecules. Incorporating GAGs into biomaterials has emerged as a widely adopted strategy in medical applications, owing to their biocompatibility and ability to control the release of bioactive molecules. Nevertheless, immobilized GAGs on biomaterials can elicit distinct cellular responses compared to their soluble forms, underscoring the need to understand the This article is protected by copyright. All rights reserved. 2 interactions between GAG and bioactive molecules within engineered functional biomaterials. By controlling critical parameters such as GAG type, density, and sulfation, it becomes possible to precisely delineate GAG functions within a biomaterial context and to better mimic specific tissue properties, enabling tailored design of GAG-based biomaterials for specific medical applications. However, this requires access to pure and well-characterized GAG compounds, which remains challenging. This review focuses on different strategies for producing well-defined GAGs and explores high-throughput approaches employed to investigate GAG-growth factor interactions and to quantify cellular responses on GAG-based biomaterials. These automated methods hold considerable promise for improving our understanding of the diverse functions of GAGs. In perspectives, we encourage the scientific community to adopt a rational approach in designing GAG-based biomaterials, taking into account the in vivo properties of the targeted tissue for medical applications

Heparan sulfate proteoglycan mediates the selective attachment and internalization of serotype 3 human adenovirus dodecahedron.

International audienceDuring adenovirus type 3 (Ad3) infection cycle, the penton (Pt) of the viral capsid, a noncovalent complex of fiber and penton base proteins, is produced in large excess and self-assembles to form a highly organized dodecahedral structure, termed dodecahedron (Dd). The physiological role of these particles is poorly understood, but we have recently reported that they can penetrate cells with high efficiency and thus may constitute an attractive tool for gene or protein delivery approaches. Surprisingly, Dd displayed the ability to enter cells non-permissive to Ad3, suggesting the existence of additional internalization modes. In this study, we show that Ad3 Dd binds to cell surface heparan sulfate (HS) through high affinity interaction with the penton base. Furthermore, binding to HS was found to be the prerequisite for a novel and Dd specific entry pathway that could not be used by Ad3. Overall, these data provide new insights in the possible role of Dd during viral infection and potential therapeutic applications

HS and Inflammation: A Potential Playground for the Sulfs?

International audienceHeparan sulfate (HS) is a complex polysaccharide abundantly found in extracellular matrices and cell surfaces. HS participates in major cellular processes, through its ability to bind and modulate a wide array of signaling proteins. HS/ligand interactions involve saccharide domains of specific sulfation pattern. Assembly of such domains is orchestrated by a complex biosynthesis machinery and their structure is further regulated at the cell surface by post-synthetic modifying enzymes. Amongst them, extracellular sulfatases of the Sulf family catalyze the selective removal of 6-O-sulfate groups, which participate in the binding of many proteins. As such, increasing interest arose on the regulation of HS biological properties by the Sulfs. However, studies of the Sulfs have so far been essentially restricted to the fields of development and tumor progression. The aim of this review is to survey recent data of the literature on the still poorly documented role of the Sulfs during inflammation, and to widen the perspectives for the study of this intriguing regulatory mechanism toward new physiopathological processes

The "in and out" of glucosamine 6-O-sulfation: the 6th sense of heparan sulfate.

International audienceThe biological properties of Heparan sulfate (HS) polysaccharides essentially rely on their ability to bind and modulate a multitude of protein ligands. These interactions involve internal oligosaccharide sequences defined by their sulfation patterns. Amongst these, the 6-O-sulfation of HS contributes significantly to the polysaccharide structural diversity and is critically involved in the binding of many proteins. HS 6-O-sulfation is catalyzed by 6-O-sulfotransferases (6OSTs) during biosynthesis, and it is further modified by the post-synthetic action of 6-O-endosulfatases (Sulfs), two enzyme families that remain poorly characterized. The aim of the present review is to summarize the contribution of 6-O-sulfates in HS structure/function relationships and to discuss the present knowledge on the complex mechanisms regulating HS 6-O-sulfation