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）是一种慢性、进行性肺部疾病，发病率极高。转化生长因子-B（TGF-β）和aVB 6整联蛋白是IPF的关键驱动因子; aVB 6对潜伏TGF-β施加机械力以有效地释放活性TGF-β。我们已经发现：1）aVB 6信号传导复合物募集粘蛋白-半乳糖凝集素-3调节模块; 2）MUC 1-半乳糖凝集素-3相互作用调节生长因子受体（GFR）聚集和运输。在体内，MUC 1和半乳糖凝集素-3促进肺纤维化，半乳糖凝集素-3靶向药物抑制TGF-β活性以抑制晚期进展。MUC 1在空间上限制整合素激活;大的糖蛋白漏斗受体进入粘连并施加压缩张力以促进整合素激活。1）确定MUC 1-半乳糖凝集素-3相互作用对aVB 6运输和配体接合的影响：aVB 6运输将使用生化内吞/再循环测定法进行分析。在所有目标中，将使用肺上皮细胞中的机械调节的糖蛋白模拟物和MUC 1胞外域/胞外域突变体来分析MUC 1压缩张力或信号传导的贡献。将在存在/不存在半乳糖凝集素-3或半乳糖凝集素-3靶向药物的情况下评估MUC 1-半乳糖凝集素-3相互作用的参与。2）分析MUC 1-半乳糖凝集素-3相互作用是否调节aVB 6介导的力施加和TGF-β激活：定量牵引力显微镜和TGF-β 1活性检测报告基因测定将证明MUC 1-半乳糖凝集素-3的生物物理和信号传导性质是否调节aVB 6介导的机械力并激活TGF-β。3）测试MUC 1-半乳糖凝集素-3相互作用是否协调aVB 6漏斗化以驱动TGF-β激活：学生将参观Paszek实验室，进行地形扫描角干涉显微镜检查，使用MUC 1压缩和aVB 6激活特异性抗体的FRET报告基因分析膜变形。4）确定MUC 1-半乳糖凝集素-3介导的aVB 6/TGF-B调节是否调节对半乳糖凝集素-3和aVB 6靶向药物的反应：器官型IPF模型将用于测试MUC 1-半乳糖凝集素-3相互作用对aVB 6依赖性疾病起始/进展的影响。临床前IPF模型将在给予半乳糖凝集素-3和aVB 6靶向药物后使用。新奇和及时性IPF发病率正在上升，迫切需要新的治疗管理途径。这个多学科项目将揭示aVB 6，MUC 1和galectin-3如何合作激活TGF-B;阐明驱动IPF发病机制和药物反应的机制。