Heat-shock protein 90 modulates goblet cell metaplasia in human airway epithelia

项目摘要/摘要 粘液高分泌和杯状细胞化生是许多肺部疾病的主要特征。不是 Th2水平低的哮喘、慢性支气管炎和囊性纤维化的杯状细胞化生有治疗方法。在……里面 相比之下，Th2高哮喘有描述良好的驱动因素(例如，IL13)和可用的治疗方法。患有疾病的患者 未经治疗的杯状细胞化生出现致残症状、频繁住院和高医疗保健 成本。本提案的总体目标是确定热休克蛋白90(HSP90)在 气道杯状细胞化生。我的中心假设是，抑制HSP90会诱导高脚杯 纤毛细胞转分化。强有力的初步数据支持这一假设：1)白介素13诱导高脚杯 人呼吸道上皮细胞化生，杯状细胞与纤毛细胞呈负相关 2)抑制HSP90可逆转IL13和IL17诱导的杯状细胞化生，而不诱导细胞 3)抑制HSP90抑制IL13刺激的呼吸道上皮细胞ERBB2/EGFR信号转导。我建议 三个具体目标： 具体目标1：呼吸道上皮中杯状细胞化生是否需要HSP90？我将致力于实现这一目标 利用人和小鼠缺失HSP90基因异构体。我将曝光基因编辑的小鼠和人类 体外培养的呼吸道细胞对IL-13或IL-17有抑制作用。 特定目标2：抑制HSP90是否诱导杯状细胞向纤毛细胞转分化？这就做 通过对暴露于IL13、IL17和HSP90抑制剂的人类呼吸道细胞进行谱系追踪来解决这一目标。 具体目标3：杯状细胞中ERBB2信号通路是否需要顶膜HSP90的定位 化生？我将通过蛋白质磷酸化和定位分析来解决这一目标。我会揭发 IL-13或IL-17刺激前后人呼吸道上皮细胞对HSP90的阻断抗体。 随着这项提议的完成，我预计将确定新的机制和治疗目标 逆转呼吸道杯状细胞化生。治疗由多种诱因(如IL13)引起的杯状细胞化生 和白介素17)可能会改善许多肺部疾病。 爱荷华大学对初级教职员工的支持由来已久。我的导师，扎布纳博士 也是一位拉丁裔医生--科学家。扎布纳博士是上皮肺生物学、囊性纤维化、 和基因疗法。他曾指导过几位K奖获得者独立并成功地从事职业生涯 内科医生-科学家。这项K奖将把我的研究技能扩展到发育生物学和病毒 向量。这些工具可能会在未来实现基因治疗，并支持治愈慢性支气管炎。

PROJECT SUMMARY / ABSTRACT Mucus hypersecretion and goblet cell metaplasia are central features of many lung diseases. No treatments exist for goblet cell metaplasia in Th2-low asthma, chronic bronchitis, and cystic fibrosis. In contrast, Th2-high asthma has well-described drivers (e.g., IL13) and available treatments. Patients with untreated goblet cell metaplasia suffer disabling symptoms, frequent hospitalizations, and high healthcare costs. The overall objective of this proposal is to determine the role of heat-shock protein 90 (HSP90) in airway goblet cell metaplasia. My central hypothesis is that inhibition of HSP90 induces goblet to ciliated cell transdifferentiation. Robust preliminary data supports this hypothesis: 1) IL13 induces goblet cell metaplasia in human airway epithelia, with a negative correlation between goblet and ciliated cell abundance, 2) HSP90 inhibition reverts IL13- and IL17-induced goblet cell metaplasia without inducing cell death, and 3) HSP90 inhibition blocks ERBB2/EGFR signaling in IL13-stimulated airway epithelia. I propose three specific aims: Specific Aim 1: Is HSP90 required for goblet cell metaplasia in airway epithelia? I will address this aim using human and mouse deletion of HSP90 gene isoforms. I will expose gene-edited mice and human airway cells in vitro to IL13 or IL17. Specific Aim 2: Does HSP90 inhibition induce transdifferentiation of goblet cells into ciliated cells? I will address this aim with lineage tracing of human airway cells exposed to IL13, IL17, and HSP90 inhibitors. Specific Aim 3: Is apical membrane HSP90 localization required for ERBB2 signaling in goblet cell metaplasia? I will address this aim with protein phosphorylation and localization assays. I will expose human airway epithelia to HSP90 blocking antibodies before and after IL13 or IL17 stimulation. With the completion of this proposal, I expect to have identified novel mechanisms and treatment targets to revert airway goblet cell metaplasia. Treating goblet cell metaplasia caused by various triggers (e.g., IL13 and IL17) may improve many lung diseases. The University of Iowa has a longstanding history of support to its junior faculty. My mentor, Dr. Zabner, is also a Latino Physician-Scientist. Dr. Zabner is a national leader in epithelial lung biology, cystic fibrosis, and gene therapy. He has mentored several K awardees to independence and successful careers as Physician-Scientists. This K-award will expand my research skills to developmental biology and viral vectors. These tools may enable gene therapy in the future and support a cure for chronic bronchitis.