Targeting lung fibrosis: Dissecting molecular crosstalk mechanisms that drive disease pathogenesis and therapeutic response

针对肺纤维化：剖析驱动疾病发病机制和治疗反应的分子串扰机制

• 批准号: 2508690
• 金额: --
• 依托单位: University of Liverpool
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2508690
• 关键词: Targeting; lung; fibrosis; Dissecting; molecular

Idiopathic pulmonary fibrosis (IPF) is chronic, progressive, lung disease with very high morbidity. Transforming growth factor-B (TGF-B) and aVB6 integrin are key drivers of IPF; aVB6 applies mechanical force on latent TGF-B to potently release active TGF-B.We have found: 1) aVB6 signalling complexes recruit a mucin-galectin-3 regulatory module; 2) MUC1-galectin-3 interaction regulates growth factor receptor (GFR) clustering and trafficking. In vivo, MUC1 and galectin-3 promote lung fibrosis and galectin-3-targeting drugs inhibit TGF-B activity to suppress late-stage progression.MUC1 spatially-constrains integrin activation; the bulky glycoprotein funnels receptors into adhesions and applies compressive tension to promote integrin activation.This new paradigm of integrin activation leads to our hypothesis: Galectin-3 and MUC1 co-ordinate aVB6 clustering to drive TGF-B activation during IPF pathogenesis.Objectives & Experimental Approach1) Determine impact of MUC1-galectin-3 interaction on aVB6 trafficking and ligand engagement:aVB6 trafficking will be analysed using biochemical endocytosis/recycling assays. aVB6 clustering and ligand-binding will be analysed by immunofluorescence using activation-specific anti-aVB6 antibodies.In all objectives, contributions of MUC1 compressive tension or signalling will be analysed using mechanically-tuned glycoprotein-mimetics and MUC1 ectodomain/cytodomain mutants in lung epithelial cells. Involvement of MUC1-galectin-3 interaction, will be assessed in presence/absence of galectin-3 or galectin-3-targeting drugs.2) Analyse whether MUC1-galectin-3 interaction regulates aVB6-medicated force application and TGF-B activation:Quantitative traction force microscopy and TGF-B1 activity luciferse reporter assays will demonstrate whether biophysical and signalling properties of MUC1-galectin-3 regulate aVB6-mediated mechanical forces and activate TGF-B.3) Test whether MUC1-galectin-3 interaction co-ordinates aVB6 funnelling to drive TGF-B activation:Student will visit Paszek Lab to perform Topographical-Scanning Angle Interference Microscopy to analyse membrane deformation using FRET reporters of MUC1 compression and aVB6 activation-specific antibodies.4) Determine whether MUC1-galectin-3-mediated regulation of aVB6/TGF-B modulates response to galectin-3- and aVB6-targeting drugs:Organotypic IPF models will be used to test involvement of MUC1-galectin-3 interaction on aVB6-dependent disease initiation/progression. Preclinical IPF models will be utilised following administration of galectin-3- and aVB6-targeting drugs.Novelty & TimelinessIPF incidence is rising and in critical need of novel avenues for therapeutic management. This multidisciplinary project will reveal how aVB6, MUC1 and galectin-3 co-operate to activate TGF-B; illuminating the mechanisms driving IPF pathogenesis and drug responses.

特发性肺纤维化（IPF）是慢性，进行性肺部疾病，发病率很高。转化生长因子-B（TGF-B）和AVB6整合素是IPF的关键驱动因素； AVB6在潜在TGF-B上施加机械力来有效释放活跃的TGF-B。我们发现：1）AVB6信号复合物募集粘蛋白 - galectin-3调节模块； 2）MUC1-GALECTIN-3相互作用调节生长因子受体（GFR）聚类和运输。在体内，MUC1和GALECTIN-3促进肺纤维化和甘油-3靶向药物抑制TGF-B活性以抑制后期的进展。笨重的糖蛋白会导致受体进入粘附中，并应用压缩张力来促进整联蛋白的激活。整联蛋白激活的新范式可导致我们的假设：Galectin-3和MUC1共同辅助AVB6聚类以驱动IPF病原体中的TGF-B激活，并在IPF病原体中驱动TGF-B激活。配体参与：将使用生化内吞作用/回收测定法对AVB6运输进行分析。 aVB6 clustering and ligand-binding will be analysed by immunofluorescence using activation-specific anti-aVB6 antibodies.In all objectives, contributions of MUC1 compressive tension or signalling will be analysed using mechanically-tuned glycoprotein-mimetics and MUC1 ectodomain/cytodomain mutants in lung epithelial cells. Involvement of MUC1-galectin-3 interaction, will be assessed in presence/absence of galectin-3 or galectin-3-targeting drugs.2) Analyse whether MUC1-galectin-3 interaction regulates aVB6-medicated force application and TGF-B activation:Quantitative traction force microscopy and TGF-B1 activity luciferse reporter assays will demonstrate whether biophysical and signalling properties of MUC1-galectin-3 regulate aVB6-mediated mechanical forces and activate TGF-B.3) Test whether MUC1-galectin-3 interaction co-ordinates aVB6 funnelling to drive TGF-B activation:Student will visit Paszek Lab to perform Topographical-Scanning Angle Interference Microscopy to analyse membrane deformation using FRET reporters of MUC1 compression and aVB6 activation-specific 4）确定MUC1-GALECTIN-3介导的AVB6/TGF-B调节是否调节对半乳糖素3-和AVB6靶向药物的反应：细胞型IPF模型将用于测试MUC1-Galectin-3相互作用的MUC1-Galectin-3相互作用。临床前IPF模型将在给药后进行半乳糖素-3-和AVB6靶向药物后使用。Novelty＆TimelinessIPF发病率正在上升，并且迫切需要用于治疗管理的新型途径。这个多学科的项目将揭示AVB6，MUC1和Galectin-3合作量如何激活TGF-B；照亮驱动IPF发病机理和药物反应的机制。