Galectin binding to cells and glycoproteins with genetically modified glycosylation reveals galectin–glycan specificities in a natural context

Nielsen, Mathias I.; Stegmayr, John; Grant, Oliver C.; Yang, Zifeng; Nilsson, Ulf J.; Boos, Irene; Carlsson, Michael C.; Woods, Robert J.; Unverzagt, Carlo; Leffler, Hakon; Wandall, Hans H.

doi:10.1074/jbc.ra118.004636

Cited by 73 publications

(65 citation statements)

References 62 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…35 The N-and C-terminal CRDs of Gal-9 have both been shown to bind with sub-to low-micromolar affinity to glycans. 39,40 We propose a model in which Gal-9 binds bivalently to TIM-3 in several different modes, only some of which M6903 can compete with. The tandem CRDs of Gal-9 can bind to two sites in one TIM-3 molecule, as shown in Figure 6, or act as a bridge between TIM-3 molecules to form multimeric complexes.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of M6903, an antagonistic anti–TIM-3 monoclonal antibody

et al. 2020

View full text Add to dashboard Cite

T cell immunoglobulin and mucin domain-3 (TIM-3) is an immune checkpoint that regulates normal immune responses but can be exploited by tumor cells to evade immune surveillance. TIM-3 is primarily expressed on immune cells, particularly on dysfunctional and exhausted T cells, and engagement of TIM-3 with its ligands promotes TIM-3-mediated T cell inhibition. Antagonistic ligand-blocking anti-TIM-3 antibodies have the potential to abrogate T cell inhibition, activate antigen-specific T cells, and enhance anti-tumor immunity. Here we describe M6903, a fully human anti-TIM-3 antibody without effector function and with high affinity and selectivity to TIM-3. We demonstrate that M6903 blocks the binding of TIM-3 to three of its ligands, phosphatidylserine (PtdSer), carcinoembryonic antigen cell adhesionrelated molecule 1 (CEACAM1), and galectin 9 (Gal-9). These results are supported by an atomic resolution crystal structure and functional assays, which demonstrate that M6903 monotherapy enhanced T cell activation. This activation was further enhanced by the combination of M6903 with bintrafusp alfa, a bifunctional fusion protein that simultaneously blocks the transforming growth factorβ (TGF-β) and programmed death ligand 1 (PD-L1) pathways. M6903 and bintrafusp alfa combination therapy also enhanced anti-tumor efficacy in huTIM-3 knock-in mice, relative to either monotherapy. These in vitro and in vivo data, along with favorable pharmacokinetics in marmoset monkeys, suggest that M6903 as a monotherapy warrants further pre-clinical assessment and that M6903 and bintrafusp alfa may be a promising combination therapy in the clinic. ARTICLE HISTORY

show abstract

Section: Discussionmentioning

confidence: 99%

Identification and characterization of M6903, an antagonistic anti–TIM-3 monoclonal antibody

et al. 2020

View full text Add to dashboard Cite

show abstract

“…(I) There were 122 overlapping proteins between the exogenous and transfection protocols (SI Appendix, Table S1). (37,38). Thus, we next focused our investigation on profiling N-glycans, with particular attention on assessing the abundance of N-glycans bearing the high-affinity terminal galactose monosaccharides (39).…”

Section: Validation Of Interactions Between Galectin-3 and Identifiedmentioning

confidence: 99%

Mapping glycan-mediated galectin-3 interactions by live cell proximity labeling

Joeh

O’Leary

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Galectin-3 is a glycan-binding protein (GBP) that binds β-galactoside glycan structures to orchestrate a variety of important biological events, including the activation of hepatic stellate cells and regulation of immune responses. While the requisite glycan epitopes needed to bind galectin-3 have long been elucidated, the cellular glycoproteins that bear these glycan signatures remain unknown. Given the importance of the three-dimensional (3D) arrangement of glycans in dictating GBP interactions, strategies that allow the identification of GBP receptors in live cells, where the native glycan presentation and glycoprotein expression are preserved, have significant advantages over static and artificial systems. Here we describe the integration of a proximity labeling method and quantitative mass spectrometry to map the glycan and glycoprotein interactors for galectin-3 in live human hepatic stellate cells and peripheral blood mononuclear cells. Understanding the identity of the glycoproteins and defining the structures of the glycans will empower efforts to design and develop selective therapeutics to mitigate galectin-3–mediated biological events.

show abstract

“…Our data on SF lubricin O-glycan truncation in OA also bears implications on binding to galectin-3. Previous studies have indicated that galectin-3 exhibits only weak affinity to Oglycoproteins and O-glycopeptides expressing single core 1 O-glycans (Galb1-3GalNAc or NeuAcα2-3Galb1-3GalNAc) (35,45).…”

Section: Functional Implications Of Lubricin Glycosylationmentioning

confidence: 99%

Decrease of core 2 O-glycans on synovial lubricin in osteoarthritis reduces galectin-3 mediated crosslinking

Flowers

Thomsson

Ali

et al. 2020

Journal of Biological Chemistry

View full text Add to dashboard Cite

The synovial fluid glycoprotein lubricin (also known as proteoglycan 4) is a mucin-type O-linked glycosylated biological lubricant implicated to be involved in osteoarthritis (OA) development. Lubricin’s ability to reduce friction is related to its glycosylation consisting of sialylated and unsialylated Tn-antigens, core-1 and core-2 structures. The glycans on lubricin have also been suggested to be involved in crosslinking and stabilization of the lubricating superficial layer of cartilage by mediating interaction between lubricin and galectin-3. However, with the spectrum of glycans being found on lubricin, the glycan candidates involved in this interaction were unknown. Here, we confirm that the core-2 O-linked glycans mediate this lubricin-galectin 3 interaction, shown by surface plasmon resonance data indicating that recombinant lubricin (rhPRG4) devoid of core-2 structures did not bind to recombinant galectin-3. Conversely, transfection of Chinese hamster ovary (CHO) cells with the core 2 GlcNAc transferase acting on a mucin type O-glycoprotein displayed increased galectin-3 binding. Both the level of galectin-3 and the galectin-3 interactions with synovial lubricin were found to be decreased in late-stage OA patients, coinciding with an increase in truncated and less sialylated core 1 O-glycans and Tn-antigens. These data suggest a defect in cross-linking of surface-active molecules in OA and provides novel insights into OA molecular pathology.

show abstract

Galectin binding to cells and glycoproteins with genetically modified glycosylation reveals galectin–glycan specificities in a natural context

Cited by 73 publications

References 62 publications

Identification and characterization of M6903, an antagonistic anti–TIM-3 monoclonal antibody

Identification and characterization of M6903, an antagonistic anti–TIM-3 monoclonal antibody

Mapping glycan-mediated galectin-3 interactions by live cell proximity labeling

Decrease of core 2 O-glycans on synovial lubricin in osteoarthritis reduces galectin-3 mediated crosslinking

Contact Info

Product

Resources

About