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Key Points Glycosylation is a key cellular mechanism regulating several physiological and pathological functions. Alterations in glycoproteins, glycosphingolipids and proteoglycans are common features of cancer cells. The most-widely occurring cancer-associated changes in protein glycosylation are increased sialylation, increased branched-glycan structures and overexpression of 'core' fucosylation. The overexpression of branched- N -glycan structures interferes with epithelial cadherin-mediated cell–cell adhesion, promoting tumour cell dissociation and invasion. Modifications of integrins with branched N -glycans, truncated O -glycans and/or sialylated structures modulate tumour cell–matrix interactions, fostering the process of tumour cell migration. Altered expression of proteoglycans and their glycosaminoglycan chains interfere with extracellular signalling molecules and modulate the activation of tyrosine kinase protein receptors. Altered glycosylation of growth factor receptors and the modified expression of gangliosides affect cancer cell signal transduction pathways, modulating tumour cell growth and proliferation. Glycans and their corresponding endogenous carbohydrate-recognition lectins are key regulators of the inflammation and immune response towards the tumour cells. Several serological markers currently used in the clinic are based on the detection of circulating glycoproteins or glycoconjugates with altered glycosylation. Glycans have major potential applications in improving early diagnosis, determination of prognosis and risk stratification, as well as in serving as markers of specific therapeutic targets. This Review discusses the importance of glycobiology in cancer research, given its role in cancer development and progression, and provides an overview of possible targets for diagnostic application and therapeutic strategies. Despite recent progress in understanding the cancer genome, there is still a relative delay in understanding the full aspects of the glycome and glycoproteome of cancer. Glycobiology has been instrumental in relevant discoveries in various biological and medical fields, and has contributed to the deciphering of several human diseases. Glycans are involved in fundamental molecular and cell biology processes occurring in cancer, such as cell signalling and communication, tumour cell dissociation and invasion, cell–matrix interactions, tumour angiogenesis, immune modulation and metastasis formation. The roles of glycans in cancer have been highlighted by the fact that alterations in glycosylation regulate the development and progression of cancer, serving as important biomarkers and providing a set of specific targets for therapeutic intervention. This Review discusses the role of glycans in fundamental mechanisms controlling cancer development and progression, and their applications in oncology.

Aspergillus fumigatus is the most common cause of invasive mold disease in humans. The mechanisms underlying the adherence of this mold to host cells and macromolecules have remained elusive. Using mutants with different adhesive properties and comparative transcriptomics, we discovered that the gene uge3, encoding a fungal epimerase, is required for adherence through mediating the synthesis of galactosaminogalactan. Galactosaminogalactan functions as the dominant adhesin of A. fumigatus and mediates adherence to plastic, fibronectin, and epithelial cells. In addition, galactosaminogalactan suppresses host inflammatory responses in vitro and in vivo, in part through masking cell wall β-glucans from recognition by dectin-1. Finally, galactosaminogalactan is essential for full virulence in two murine models of invasive aspergillosis. Collectively these data establish a role for galactosaminogalactan as a pivotal bifunctional virulence factor in the pathogenesis of invasive aspergillosis.

The galactosaminogalactan (GAG) is a cell wall component of Aspergillus fumigatus that has potent anti-inflammatory effects in mice. However, the mechanisms responsible for the anti-inflammatory property of GAG remain to be elucidated. In the present study we used in vitro PBMC stimulation assays to demonstrate, that GAG inhibits proinflammatory T-helper (Th)1 and Th17 cytokine production in human PBMCs by inducing Interleukin-1 receptor antagonist (IL-1Ra), a potent anti-inflammatory cytokine that blocks IL-1 signalling. GAG cannot suppress human T-helper cytokine production in the presence of neutralizing antibodies against IL-1Ra. In a mouse model of invasive aspergillosis, GAG induces IL-1Ra in vivo, and the increased susceptibility to invasive aspergillosis in the presence of GAG in wild type mice is not observed in mice deficient for IL-1Ra. Additionally, we demonstrate that the capacity of GAG to induce IL-1Ra could also be used for treatment of inflammatory diseases, as GAG was able to reduce severity of an experimental model of allergic aspergillosis, and in a murine DSS-induced colitis model. In the setting of invasive aspergillosis, GAG has a significant immunomodulatory function by inducing IL-1Ra and notably IL-1Ra knockout mice are completely protected to invasive pulmonary aspergillosis. This opens new treatment strategies that target IL-1Ra in the setting of acute invasive fungal infection. However, the observation that GAG can also protect mice from allergy and colitis makes GAG or a derivative structure of GAG a potential treatment compound for IL-1 driven inflammatory diseases.

Existing technologies employed to generate antibodies against bacterial polysaccharides and proteins rely on the availability of purified or synthetic antigens. Here, we present a genetics-based platform that utilises Citrobacter rodentium (CR), an enteric mouse pathogen, to both produce and present complex heterologous polysaccharides and protein antigen complexes during natural infection. As proof of concept, we use lipopolysaccharides (O), capsular polysaccharides (K) and type 3 fimbrial (T3F) antigens expressed by the WHO critical priority pathogens Klebsiella pneumoniae (KP) and Escherichia coli (EC). Following one infection cycle (28 days), CR induces specific IgG antibodies against KPO1, ECO25b, KPK2 and KPT3F. We demonstrate that the antibodies are functional in downstream applications, including protection against pathogenic KP challenge, KP capsular serotyping and KP biofilm inhibition. Whilst KP and EC antigens were used as prototypical examples, this modular platform is now readily adaptable to generate antibodies against diverse polysaccharide and protein antigens, with basic science, public health and therapeutic applications. Bacterial antigens, such as lipopolysaccharides, are complex structures which remain difficult to synthesise or purify for antibody generation. Here, authors present a platform technology using Citrobacter rodentium - an enteric mouse pathogen - to both produce and present complex antigens for antibody generation.

Streptococcus uberis is a causative pathogen of bovine mastitis with high genetic diversity. Rhamnose-rich polysaccharides (RPS) are abundant surface structures covalently anchored to peptidoglycan and represent promising vaccine candidates for several streptococcal pathogens. It was previously reported that the RPS of S. uberis strain 233 is composed of a repeating → 2)-α- l -Rha p -(1 → 3)-α- l -Rha p -(1 → disaccharide backbone decorated with α- d -Glc p side-chains. In this study, we identified a hitherto unknown glycerol phosphate (GroP) modification at the 6-OH of the Glc residue in S. uberis 233 RPS using nuclear magnetic resonance analysis. Comparative genomic analysis of 592 S. uberis genomes revealed significant diversity in the RPS biosynthesis gene cluster with six major RPS genotypes. RPS genotypes 1–4, representing 97.5% of the analyzed strains, all contained the rhamnan backbone biosynthesis genes shared between several streptococcal species, as well as a putative GroP transferase gene. Using rhamnan-reactive immune serum, we further demonstrated that rhamnan is a conserved and accessible glycan motif in S. uberis RPS genotype 1 and 2 strains, but this motif is inferred to be shielded by side-chains in genotype 4 strains. Importantly, experiments with sera from cattle, challenged intramammarily with S. uberis , revealed that the rhamnan backbone of S. uberis RPS is an immunogenic glycan motif and remained accessible to bovine IgG antibodies in the presence of single residue RPS side-chains. Overall, this study suggests that S. uberis RPS are modified with GroP and reports that RPS in most strains contain a conserved, immunogenic and antibody accessible rhamnan glycan motif.

Carbohydrates are commonly found in conjunction with lipids or proteins, resulting in the formation of glycoconjugates such as glycoproteins, glycolipids, and proteoglycans. These glycoconjugates are essential in various biological activities, including inflammation, cell-cell recognition, bacterial infections, and immune response. Nonetheless, the isolation of naturally occurring glycoconjugates presents challenges due to their typically heterogeneous nature, resulting in variations between batches in structure and function, impeding a comprehensive understanding of their mechanisms of action. Consequently, there is a strong need for the efficient synthesis of artificial glycoconjugates with precisely described compositions and consistent biological properties. The chemical and enzymatic approaches discussed in this paper present numerous research opportunities to develop customised glycoconjugate vaccines.

Glycoconjugate vaccines provide effective prophylaxis against bacterial infections. To date, however, no commercial vaccine has been available in which the key carbohydrate antigens are produced synthetically. We describe the large-scale synthesis, pharmaceutical development, and clinical evaluation of a conjugate vaccine composed of a synthetic capsular polysaccharide antigen of Haemophilus influenzae type b (Hib). The vaccine was evaluated in clinical trials in Cuba and showed long-term protective antibody titers that compared favorably to licensed products prepared with the Hib polysaccharide extracted from bacteria. This demonstrates that access to synthetic complex carbohydrate-based vaccines is feasible and provides a basis for further development of similar approaches for other human pathogens.

Severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS) coronaviruses (CoVs) are zoonotic pathogens with high fatality rates and pandemic potential. Vaccine development focuses on the principal target of the neutralizing humoral immune response, the spike (S) glycoprotein. Coronavirus S proteins are extensively glycosylated, encoding around 66–87 N-linked glycosylation sites per trimeric spike. Here, we reveal a specific area of high glycan density on MERS S that results in the formation of oligomannose-type glycan clusters, which were absent on SARS and HKU1 CoVs. We provide a comparison of the global glycan density of coronavirus spikes with other viral proteins including HIV-1 envelope, Lassa virus glycoprotein complex, and influenza hemagglutinin, where glycosylation plays a known role in shielding immunogenic epitopes. Overall, our data reveal how organisation of glycosylation across class I viral fusion proteins influence not only individual glycan compositions but also the immunological pressure across the protein surface. Glycosylation plays a key role in shielding of immunogenic epitopes on viral spike (S) proteins. Here Watanabe et al. report that glycans of coronavirus SARS and MERS S proteins are heterogeneously distributed and do not form an efficacious high-density global shield which would ensure efficient immune evasion.

Scientists are unpicking the life cycle of SARS-CoV-2 and how the virus uses tricks to evade detection. Scientists are unpicking the life cycle of SARS-CoV-2 and how the virus uses tricks to evade detection. Credit: Janet Iwasa, University of Utah Rendered impression of SARS-CoV-2

In vaccine design, antigens are often arrayed in a multivalent nanoparticle form, but in vivo mechanisms underlying the enhanced immunity elicited by such vaccines remain poorly understood. We compared the fates of two different heavily glycosylated HIV antigens, a gp120-derived mini-protein and a large, stabilized envelope trimer, in protein nanoparticle or “free” forms after primary immunization. Unlike monomeric antigens, nanoparticles were rapidly shuttled to the follicular dendritic cell (FDC) network and then concentrated in germinal centers in a complement-, mannose-binding lectin (MBL)–, and immunogen glycan–dependent manner. Loss of FDC localization in MBL-deficient mice or via immunogen deglycosylation significantly affected antibody responses. These findings identify an innate immune–mediated recognition pathway promoting antibody responses to particulate antigens, with broad implications for humoral immunity and vaccine design.