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

Joeh, Eugene; O’Leary, Timothy R.; Li, Weichao; Hawkins, Richard L.; Hung, Jonathan R.; Parker, Christopher G.; Huang, Mia L.

doi:10.1073/pnas.2009206117

Cited by 44 publications

(40 citation statements)

References 63 publications

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“…Gal3 has been implicated in numerous pathologies [15,31], is present at different subcellular locations [32][33][34][35][36][37] and has almost 250 cell-surface binding partners [38]. Because of this, the exact mechanism by which stromal produced extracellular Gal3 protects BCP-ALL cells against the conventional chemotherapy drugs vincristine and nilotinib, as found in our study, is difficult to determine.…”

Section: Discussionmentioning

confidence: 75%

Overcoming Microenvironment-Mediated Chemoprotection through Stromal Galectin-3 Inhibition in Acute Lymphoblastic Leukemia

Tarighat

Fei

Joo

et al. 2021

IJMS

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Environmentally-mediated drug resistance in B-cell precursor acute lymphoblastic leukemia (BCP-ALL) significantly contributes to relapse. Stromal cells in the bone marrow environment protect leukemia cells by secretion of chemokines as cues for BCP-ALL migration towards, and adhesion to, stroma. Stromal cells and BCP-ALL cells communicate through stromal galectin-3. Here, we investigated the significance of stromal galectin-3 to BCP-ALL cells. We used CRISPR/Cas9 genome editing to ablate galectin-3 in stromal cells and found that galectin-3 is dispensable for steady-state BCP-ALL proliferation and viability. However, efficient leukemia migration and adhesion to stromal cells are significantly dependent on stromal galectin-3. Importantly, the loss of stromal galectin-3 production sensitized BCP-ALL cells to conventional chemotherapy. We therefore tested novel carbohydrate-based small molecule compounds (Cpd14 and Cpd17) with high specificity for galectin-3. Consistent with results obtained using galectin-3-knockout stromal cells, treatment of stromal-BCP-ALL co-cultures inhibited BCP-ALL migration and adhesion. Moreover, these compounds induced anti-leukemic responses in BCP-ALL cells, including a dose-dependent reduction of viability and proliferation, the induction of apoptosis and, importantly, the inhibition of drug resistance. Collectively, these findings indicate galectin-3 regulates BCP-ALL cell responses to chemotherapy through the interactions between leukemia cells and the stroma, and show that a combination of galectin-3 inhibition with conventional drugs can sensitize the leukemia cells to chemotherapy.

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Section: Discussionmentioning

confidence: 75%

Overcoming Microenvironment-Mediated Chemoprotection through Stromal Galectin-3 Inhibition in Acute Lymphoblastic Leukemia

Tarighat

Fei

Joo

et al. 2021

IJMS

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“…As such, attention to the physiological context under which the function of sugar-binding proteins are studied is crucial. A recent study from the Huang group examined the glycosylation-dependent interactions of galectins with intracellular and surface expressed ligands through proximal labeling of cells with galectin-fusion proteins (59). The application of this technique to ECs exposed to inflammatory mediators and/or culture under fluid shear stress could be extremely valuable for understanding the mechanisms and regulators of galectin-glycan interactions in context.…”

Section: Discussionmentioning

confidence: 99%

Vascular Endothelial Galectins in Leukocyte Trafficking

2021

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Leukocyte recruitment to the site of injury is a crucial event in the regulation of an inflammatory response. Tight regulation of interactions between the endothelium and circulating leukocytes is necessary to ensure a protective response to injury does not result in inflammatory disease. Rising interest in the broad immunoregulatory roles displayed by members of the glycan-binding galectin family suggests that these proteins could be an attractive target for therapeutic intervention, since their expression is significantly altered in disease. The focus of this review is to summarize current knowledge on the role of galectins in leukocyte trafficking during inflammation and the clinical approaches being taken to target these interactions for treatment of inflammatory disease.

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“…Live cell proximity labeling. Proximity tagging was adapted from previously published procedures (44,65). mESCs seeded on gelatin were treated with cholPEGNTA (10 μM, 1 hr, 37°C) and further incubated with APEX2 fused proteoglycans (10 μM, 1 hr, 37°C) after washing 2X DPBS.…”

Section: Methodsmentioning

confidence: 99%

“…The biotinylated proteins can then be probed using fluorophore-conjugated streptavidin and imaged using fluorescence microscopy or enriched for mass spectrometry-based identification. The APEX2 approach is especially apt for glycan-protein interactions that are heavily influenced by three-dimensional presentations of ligands, and which may be dynamic or weak (44).…”

Section: Metabolic Engineering With Azido Xylosides To Hijack Gag Production To Generatementioning

confidence: 99%

Chemical Editing of Proteoglycan Architecture

O’Leary¹,

Critcher²,

Stephenson³

et al. 2021

Preprint

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Proteoglycans are heterogeneous macromolecular glycoconjugates that orchestrate many important cellular processes. While much attention has focused on the poly-sulfated glycosaminoglycan chains that decorate proteoglycans, other important elements of proteoglycan architecture, such as their core proteins and cell surface localization, have garnered less emphasis. Hence, comprehensive structure-function relationships that consider the replete proteoglycan architecture as glycoconjugates are limited. Here, we present a comprehensive approach to study proteoglycan structure and biology by fabricating defined semi-synthetic modular proteoglycans that can be tailored for cell surface display. To do so, we integrate amber codon reassignment in the expression of sequence-fined proteoglycan core proteins, metabolic oligosaccharide engineering to produce functionalizable glycosaminoglycans, and bioorthogonal click chemistry to covalently tether the two components. These materials permit the methodical dissection of the parameters required for optimal binding and function of various proteoglycan-binding proteins, and they can be modularly displayed on the surface of any living cell. We demonstrate that these sophisticated materials can recapitulate the functions of native proteoglycans in mouse embryonic stem cell differentiation and cancer cell spreading, while permitting the identification of the most important contributing elements of proteoglycan architecture toward function. This technology platform will confer structural resolution toward the investigation of proteoglycan structure-function relationships in cell biology.

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

Cited by 44 publications

References 63 publications

Overcoming Microenvironment-Mediated Chemoprotection through Stromal Galectin-3 Inhibition in Acute Lymphoblastic Leukemia

Overcoming Microenvironment-Mediated Chemoprotection through Stromal Galectin-3 Inhibition in Acute Lymphoblastic Leukemia

Vascular Endothelial Galectins in Leukocyte Trafficking

Chemical Editing of Proteoglycan Architecture

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