Mucin sialylation drives epithelial cell senescence and severe asthma

粘蛋白唾液酸化导致上皮细胞衰老和严重哮喘

基本信息

批准号：
10026633
负责人：
Richard Charles Boucher
金额：
$ 74.68万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10026633
关键词：
Agonist Asthma Biological Biological Markers CCND1 gene Carbohydrates Cell Aging Cell Communication Cell Count Cell Culture Techniques Cell Differentiation process Cell surface Cells Chronic Data Disease ERBB2 gene Epidermal Growth Factor Receptor Epithelial Epithelial Cell Proliferation Epithelial Cells Epithelium Evolution Family Family member Functional disorder Galactosides Gene Expression Genes Glutathione Glutathione Disulfide Goblet Cells Human Human Biology Hyperplasia In Vitro Inflammation Interleukin-13 Link Lung Measures Metabolic Metabolic dysfunction Metabolic stress Metabolism Mitochondria Mucins Mus Oxidative Stress Pathway interactions Phenotype Post-Translational Protein Processing Process Proteins Proto-Oncogene Proteins c-akt Publishing Pyroglyphidae Reporting Role Severities Severity of illness Sialyltransferases Sputum Stimulus TNF gene Testing Time Tissues Transgenic Mice Up-Regulation airway epithelium asthma exacerbation asthmatic asthmatic airway asthmatic patient cell type driving force epithelial wound extracellular glycosylation improved in vivo mTOR inhibition mouse model new therapeutic target novel oxidation particle protein expression receptor senescence sialylation wound healing

项目摘要

This application explores the paradigm shifting hypothesis that post-translational modification (sialylation) of a cell surface (tethered) mucin, MUC4, drives terminal differentiation and senescence of airway epithelial cells (AECs) through inhibition of epidermal growth factor receptor (EGFR) family pathways, worsening epithelial wound repair and asthma severity. Our proposed studies will be the first to specifically test tethered mucins and their post-translational N-glycosylation/sialylation for a role in AEC terminal differentiation and senescence. Our published data (Zhou et al) demonstrate that sialylation of the tethered mucin, MUC4β, i.e., MUC4α βSA, controls goblet cell terminal differentiation as a function of β-galactoside α2,6-sialyltransferase-1 (ST6GAL1) activity. Both ST6GAL1 and MUC4β critically lower AEC proliferative capacity, with additional published data (Inoue et al) linking diminished AEC proliferation to abnormal wound repair in vitro. Reduced AEC proliferative capacity may reflect reduced activation of the EGFR family member, ErbB2, a known receptor for MUC4β. New data confirm that elevations in MUC4 and ST6GAL1 are present in severe exacerbation-prone asthma and in house dust mite challenged mice. Elevations in both biologically associate with low intracellular glutathione (GSH) to oxidized glutathione (GSSG) ratios (oxidative stress) and lower mitochondrial and senescence gene expression. Parallel in vitro AEC data show T2 inflammation/IL-13 alters intracellular metabolism and mitochondria, decreases GSH/GSSG, and further decreases wound repair. Yet, the overall impact of MUC4β and/or its sialylation to epithelial cell phenotypes, cell senescence, and the mechanisms by which these changes contribute to severe asthma are unclear. The studies proposed here will comprehensively evaluate the intersection of mucins, mucin sialylation, and senescence pathway(s), while robustly testing their functional importance using primary human AEC cell cultures and transgenic mouse models. The three proposed aims will: 1) elucidate the mechanistic and functional impact of MUC4β and MUC4βSA on AEC terminal differentiation, senescence and wound repair in vitro, 2) test the functional impact of Muc4βSA-AEC interactions on mucin secretion, goblet cell hyperplasia/terminal differentiation, senescence, and wound repair in an asthma mouse model and 3) define the relationship of MUC4βSA to senescence and AEC phenotypes in human asthma patients. These concurrent aims will iteratively develop data that link mucins, their sialylation and fundamental senescence-related epithelial processes to identify highly novel targets for treatment of severe asthma.

该应用程序探讨了范式转移假设，即翻译后修饰（溶解）细胞表面（束缚）粘蛋白MUC4驱动末端分化和气道上皮细胞的感应（AEC）通过抑制表皮生长因子受体（EGFR）家族途径，令人担忧的上皮伤口修复和哮喘严重程度。我们提出的研究将是第一个专门测试束缚的粘蛋白和他们的翻译后N-糖基化/溶解度在AEC末端分化和感应中扮演角色。我们的已发表的数据（Zhou等人）表明，链状粘蛋白MUC4β的溶解度，即MUC4αβSA，对照杯状细胞末端分化与β-半乳糖苷α2,6-溶解酶-1（ST6GAL1）活性的关系。 ST6GAL1和MUC4β批准降低了AEC增殖能力，并具有其他已发表的数据（INOUE ET） Al）将AEC增殖与体外异常伤口修复相关联。降低了AEC增殖能力可能反映了EGFR家族成员ERBB2的激活降低，ERBB2是MUC4β的已知受体。新数据确认MUC4和ST6GAL1的高程存在于严重恶化的哮喘中和内部尘螨挑战小鼠。两种生物学上的升高都与低细胞内谷胱甘肽（GSH）相关联氧化的谷胱甘肽（GSSG）比（氧化应激）和较低的线粒体和感应基因表达。平行于体外AEC数据显示T2注射/IL-13改变了细胞内代谢和线粒体，减少GSH/GSSG，并进一步减少伤口修复。然而，MUC4β和/或ITS的总体影响对上皮细胞表型，细胞感应和这些变化的机制有助于严重的哮喘尚不清楚。这里提出的研究将彻底评估粘蛋白，粘蛋白溶解和感应途径（S），同时使用使用的功能重要性原代人AEC细胞培养和转基因小鼠模型。提出的三个目标将：1）阐明 MUC4β和MUC4βSA对AEC终端分化，敏感和在体外伤口修复，2）测试MUC4βSA-AEC相互作用对粘蛋白分泌，杯状细胞的功能影响哮喘小鼠模型中的增生/末端分化，敏感和伤口修复，3）定义人哮喘患者中MUC4βSA与敏感性和AEC表型的关系。这些并发目标会迭代地开发将粘蛋白，其糖化和基本感应相关的上皮链接的数据确定严重哮喘治疗高度新颖的靶标的过程。