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

项目概要/摘要 粘液分泌过多和杯状细胞化生是许多肺部疾病的中心特征。没有 存在对Th 2-低哮喘、慢性支气管炎和囊性纤维化中杯状细胞化生的治疗。在 相反，Th 2-高哮喘具有充分描述的驱动因素（例如，IL 13）和可用的治疗。患者 未经治疗的杯状细胞化生遭受致残症状，频繁住院，和高医疗保健 成本本研究的总体目标是确定热休克蛋白90（HSP 90）在 气道杯状细胞化生。我的中心假设是，HSP 90的抑制诱导Goblet， 纤毛细胞转分化强有力的初步数据支持这一假设：1）IL 13诱导杯状 人气道上皮细胞化生，杯状细胞与纤毛细胞呈负相关 2）HSP 90抑制逆转IL 13和IL 17诱导的杯状细胞化生，而不诱导细胞凋亡。 3）HSP 90抑制阻断IL 13刺激的气道上皮中的ERBB 2/EGFR信号传导。我提议 三个具体目标： 特定目的1：气道上皮杯状细胞化生是否需要HSP 90？我将阐述这一目标 使用人类和小鼠HSP 90基因同种型的缺失。我将揭露基因编辑的老鼠和人类 体外气道细胞对IL 13或IL 17。 特异性目的2：抑制HSP 90是否诱导杯状细胞向纤毛细胞转分化？我会 通过对暴露于IL 13、IL 17和HSP 90抑制剂的人气道细胞的谱系追踪来解决这一目标。 特定目的3：杯状细胞中ERBB 2信号传导需要顶膜HSP 90定位吗 化生？我将通过蛋白磷酸化和定位分析来解决这个问题。我要揭露 人气道上皮细胞在IL 13或IL 17刺激之前和之后对HSP 90阻断抗体的反应。 随着这一提案的完成，我预计将确定新的机制和治疗目标 逆转气道杯状细胞化生治疗由各种触发因素引起的杯状细胞化生（例如，IL13 和IL 17）可以改善许多肺部疾病。 爱荷华州的大学有一个支持其初级教师的悠久历史。我的导师扎布纳博士 他也是一名拉丁裔科学家。Zabner博士是肺上皮生物学、囊性纤维化、 和基因治疗。他指导了几位K奖获得者独立和成功的职业生涯， 物理科学家这个K奖将扩大我的研究技能，发育生物学和病毒 向量。这些工具可能使基因治疗在未来和支持治疗慢性支气管炎。

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.