Journal: bioRxiv

Article Title: Targeting interactions between the Galectin-3 intrinsically disordered and structured domains based on long time-scale accelerated molecular dynamics

doi: 10.1101/2021.09.27.461147

Figure Lengend Snippet: Representative bright field images of suspensions of pediatric pre-B acute lymphoblastic leukemia LAX56 cells. Cells were incubated for 2 hours (A) with no added protein (B) with control GST, or (C - G) with GST-Gal3. (D) Addition of 100 μM TD139 Galectin-3 inhibitor. (E-H) Peptides as indicated were added together with GST or GST-Gal3. Similar results obtained were obtained with two independently generated batches of GST-Gal3; representative images are shown. (I) Quantification of agglutination expressed as the number of cell aggregates. Agglutination is defined as aggregates containing >10 cells per cluster. Error bars, mean ±SEM of cell cluster counts from 5-13 images from different areas per condition.

Article Snippet: TD139 was purchased from MedChemExpress and used at 100 μM.

Techniques: Incubation, Control, Generated, Agglutination

Journal: The Journal of Biological Chemistry

Article Title: Defining the mechanism of galectin-3–mediated TGF-β1 activation and its role in lung fibrosis

doi: 10.1016/j.jbc.2024.107300

Figure Lengend Snippet: Galectin-3 induces TGF-β1 activation in fibroblasts. Representative Western blot of pSmad2 levels in ( A ) non-IPF HLFs (N = 3) and ( B ) iHBECs (N = 2) pretreated with 50 μM SB-431542 (ALK5 inhibitor) or 1 μM GB0139 (galectin-3 inhibitor) for 20 min prior to 2-h treatment with 10 μg/ml galectin-3 or 2 ng/ml TGF-β1. Western blot bands were quantified using densitometry analysis and presented as a ratio of pSmad2/tSmad2. ALK, activin receptor–like kinase; HLF, human lung fibroblast; iHBEC, immortalized human bronchial epithelial cell line; IPF, idiopathic pulmonary fibrosis.; TGF-β1, transforming growth factor-β1.

Article Snippet: LPA-induced Smad2 phosphorylation in HLFs was also inhibited by pretreatment both with the inhaled galectin-3 inhibitor, GB0139 ( B ), and by the orally active galectin-3 inhibitors, GB1107 and GB1211 ( C ).

Techniques: Activation Assay, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Defining the mechanism of galectin-3–mediated TGF-β1 activation and its role in lung fibrosis

doi: 10.1016/j.jbc.2024.107300

Figure Lengend Snippet: Integrin-mediated TGF-β1 activation requires extracellular galectin-3. Representative Western blots of pSmad2 levels in non-IPF HLFs (N = 3) pre-treated with ( A ) NOTT199SS β1 inhibitor (0.1–100 nM) or ( B – D ) galectin-3 inhibitors GB0139, GB1107, and GB1211 (1 μM) or GB0149 (0.1–10 μM) for 20 min prior to stimulation with 2 ng/ml TGF-β1 (2-h) or 50 μM LPA (4-h). Cells pretreated with 50 μM SB-431542 (ALK5 inhibitor) were included as a control demonstrating maximal inhibition of pSmad2 signaling. Western blot bands were quantified using densitometry analysis and presented as a ratio of pSmad2/tSmad2. ALK, activin receptor–like kinase; HLF, human lung fibroblast; iHBEC, immortalized human bronchial epithelial cell line; IPF, idiopathic pulmonary fibrosis; LPA, lysophosphatidic acid; TGF-β1, transforming growth factor-β1.

Article Snippet: LPA-induced Smad2 phosphorylation in HLFs was also inhibited by pretreatment both with the inhaled galectin-3 inhibitor, GB0139 ( B ), and by the orally active galectin-3 inhibitors, GB1107 and GB1211 ( C ).

Techniques: Activation Assay, Western Blot, Control, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Defining the mechanism of galectin-3–mediated TGF-β1 activation and its role in lung fibrosis

doi: 10.1016/j.jbc.2024.107300

Figure Lengend Snippet: Galectin-3 inhibitors block αv integrin and TGFβRII binding to galectin-3. Solution competition binding assays performed with the galectin-3 inhibitor GB0139 ( blue ) or GB1107 ( black ) for αv integrins: ( A ) αvβ1, ( B ) αvβ5, and ( C ) αvβ6 or ( D ) TGFβRII in the presence of galectin-3 at 625 nM. Response values are normalized with respect to the highest binding response (DMSO control) and competitive inhibition graphs plotted in GraphPad Prism. IC 50 values were calculated by nonlinear regression analysis (binding saturation), and specific binding was calculated with hill slope. DMSO, dimethyl sulfoxide; TGF, transforming growth factor.

Article Snippet: LPA-induced Smad2 phosphorylation in HLFs was also inhibited by pretreatment both with the inhaled galectin-3 inhibitor, GB0139 ( B ), and by the orally active galectin-3 inhibitors, GB1107 and GB1211 ( C ).

Techniques: Blocking Assay, Binding Assay, Control, Inhibition

Journal: The Journal of Biological Chemistry

Article Title: Defining the mechanism of galectin-3–mediated TGF-β1 activation and its role in lung fibrosis

doi: 10.1016/j.jbc.2024.107300

Figure Lengend Snippet: Galectin-3 inhibitors reduce secretion of fibrotic markers in IPF lung tissue. The daily average change in the level of fibrotic markers secreted by IPF patient–derived PCLuS was measured in response to treatment with inhibitor. PCLuS were treated with galectin-3 inhibitor GB0139 (0.3–10 μM), 10 μM SB-525334, 2.5 mM pirfenidone, or 2.5 μM nintedanib for 4 days with media replaced every 24 h. Levels of secreted galectin-3, Col1a1 ( A ), TIMP1 ( B ), hyaluronan ( C ), and MMP7 ( D ) were measured daily, and the average value for each mediator (N = 3 donors) is presented as % change in drug treated samples compared with vehicle control. Comparative levels of galectin-3 are shown in each graph. IPF, idiopathic pulmonary fibrosis; PCLuS, precision cut lung slice.

Article Snippet: LPA-induced Smad2 phosphorylation in HLFs was also inhibited by pretreatment both with the inhaled galectin-3 inhibitor, GB0139 ( B ), and by the orally active galectin-3 inhibitors, GB1107 and GB1211 ( C ).

Techniques: Derivative Assay, Control

Journal: The Journal of Biological Chemistry

Article Title: Defining the mechanism of galectin-3–mediated TGF-β1 activation and its role in lung fibrosis

doi: 10.1016/j.jbc.2024.107300

Figure Lengend Snippet: Diagram of the proposed mechanism of galectin-3–mediated TGF-β1 activation in lung fibroblasts. Activation of αvβ1 integrins leads to a conformational change, which promotes release of the active TGFβ molecule from the large latent TGFβ complex and triggers downstream signaling following binding to TGF-β1 receptors on neighboring cells. In the absence of galectin-3 ( A ), the αvβ1 integrin and TGF-β1 receptors may not always be found in close proximity on adjacent cells, limiting the degree of TGF-β1 receptor mediated cell signaling initiated. B , in the presence of galectin-3, the galectin-3 carbohydrate-binding domain binds to the glycosylation sites on both the αv integrins and the TGF-β1 receptor forming a galectin lattice at the cell surface. This galectin lattice facilitates receptor clustering, bringing the integrin and TGF-β1 receptor into close proximity, and ensuring that the active TGFβ1 molecule can interact with its receptor potentiating TGF-β1 downstream signaling and phosphorylation of Smad2 and Smad3. Galectin inhibitors including GB0139 bind to the galectin-3 carbohydrate recognition domain, block the protein–glycan interactions and prevent galectin lattice–induced clustering. TGF-β1, transforming growth factor-β1.

Article Snippet: LPA-induced Smad2 phosphorylation in HLFs was also inhibited by pretreatment both with the inhaled galectin-3 inhibitor, GB0139 ( B ), and by the orally active galectin-3 inhibitors, GB1107 and GB1211 ( C ).

Techniques: Activation Assay, Binding Assay, Glycoproteomics, Phospho-proteomics, Blocking Assay

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Patient Demographics and Disease Characteristics at Study Entry

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques:

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Adverse Events in the Safety Population

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques:

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Trough (Before Dose) Plasma GB0139 Concentrations (ng/ml) in Patients with COVID-19

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques: Clinical Proteomics

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Individual patient serum galectin-3 concentrations from all patients receiving GB0139 + SoC (standard of care) (red) or SoC (blue). Linear regression analysis showing the line of best fit with a 95% confidence interval (shaded area). Of note, overall treatment effect, using all available data to Day 16 was also significant for GB0139 + SoC versus SoC ( post hoc analysis of covariance [ANCOVA] over days 2–16: P = 0.0001); however, the last day for data availability in the SoC arm was Day 14.

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques:

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Absolute change from baseline in markers of inflammation in the overall population: ( A ) C-reactive protein, ( B ) lactate dehydrogenase, ( C ) neutrophil/lymphocyte ratio, and ( D ) CXCL10. Individual patient-degree data from all those who received GB0139 + SoC (standard of care) (red) or SoC (blue): linear regression analysis showing the line of best fit with 95% confidence interval (shaded area). CXCL10 = C-X-C motif chemokine ligand 10.

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques:

Journal: American Journal of Respiratory and Critical Care Medicine

Article Title: An Inhaled Galectin-3 Inhibitor in COVID-19 Pneumonitis: A Phase Ib/IIa Randomized Controlled Clinical Trial (DEFINE)

doi: 10.1164/rccm.202203-0477OC

Figure Lengend Snippet: Effect of GB0139 + SoC (standard of care) (blue) and SoC (red) on coagulopathy: individual patient-degree data for ( A ) D-dimer concentrations, ( B ) fibrinogen/platelet ratio, ( C ) platelets, and ( D ) activated partial thromboplastin time. ( A ) and ( B ) display linear regression analysis showing the line of best fit with a 95% confidence interval (shaded area).

Article Snippet: GB0139 is being developed by Galecto Biotech.

Techniques: