Transcriptional regulation of goblet cell metaplasia

杯状细胞化生的转录调控

基本信息

批准号：
9279216
负责人：
Vladimir Kalinichenko
金额：
$ 39万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-05 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9279216
关键词：
ARG2 gene Allergens Applications Grants Asthma Binding Biological Assay Cell Differentiation process Chronic Chronic Obstructive Airway Disease Clara cell Clinical Cystic Fibrosis Data Disease Doxycycline Embryonic Development Epithelial Epithelial Cells FOXM1 gene Family Gene Expression Gene Targeting Genes Genetic Genetic Transcription Goals Goblet Cells Human Immunohistochemistry In Vitro Interleukin-13 Interleukin-13 Overexpression Laboratories Lasers Lung Inflammation Malignant Neoplasms Mediating Mediator of activation protein Metaplasia Modeling Molecular Morbidity - disease rate Mucous body substance Mus Pathogenesis Pathogenicity Pathway interactions Patients Peptides Pharmacology Phenotype Play Production Promoter Regions Proteins Pyroglyphidae Regulation Regulatory Element Reporter Reverse Transcriptase Polymerase Chain Reaction Role STAT6 gene Signal Pathway Signal Transduction Testing Therapeutic Time Transcriptional Regulation Transgenes Transgenic Mice Up-Regulation Western Blotting airway epithelium airway hyperresponsiveness chromatin immunoprecipitation efficacy testing experimental study forkhead protein gain of function genetic approach in vivo inhibitor/antagonist knock-down loss of function lung injury methacholine mortality mouse model new therapeutic target novel prevent promoter public health relevance response small hairpin RNA small molecule small molecule inhibitor

项目摘要

DESCRIPTION (provided by applicant): Goblet cell metaplasia, associated with increased mucus production, is a key pathogenic feature of chronic airway disorders, such as asthma, COPD and cystic fibrosis, all of which contribute to significant morbidity and mortality worldwide. Pharmacological targeting of goblet cell metaplasia represents a significant clinical challenge. Therefore, identification of new molecular mechanisms in airway epithelial differentiation will provide novel therapeutic targets for treatment of chronic airway disorders. Our grant proposal focuses on novel molecular mechanisms in goblet cell differentiation that are regulated by FoxM1, a transcription factor from the Forkhead box (FOX) family. While FoxM1 plays an important role in embryonic development and pathogenesis of various cancers, the role of FoxM1 in chronic airway diseases is unknown. In our preliminary data, FoxM1 was induced in airway epithelial cells of mice with asthma-like diseases caused by OVA, IL-13 and house dust mite extract (HDM). FoxM1 was also induced in airway epithelial cells of human patients with asthma and COPD. Genetic deletion of the Foxm1 gene (CCSP-Cre) or pharmacological inhibition of the FoxM1 protein (ARF peptide) in HDM-challenged airway epithelium effectively diminished goblet cell metaplasia, reduced lung inflammation and decreased airway hyper-responsiveness to methacholine. While these data suggest that FoxM1 plays a key role in asthma pathogenesis, molecular mechanisms regulated by FoxM1 remain uncharacterized. We propose to test the hypothesis that FoxM1 acts downstream of the IL-13/Stat6 pathway to induce expression of goblet cell-specific genes in airway epithelial cells. In Aim I, we will use transgenic mice with FoxM1 "gain-of-function" and "loss-of-function" in airway Clara cells to identify downstream FoxM1 target genes critical for allergen-mediated differentiation of Clara cells into goblet cells. Furthermore, we provide preliminary data demonstrating that IL-13 induces FoxM1 expression in cultured human airway epithelial cells and airway epithelium of transgenic mice. Knockdown of FoxM1 in vitro inhibited differentiation of airway epithelial cells toward goblet cell phenotype in response to IL-13 stimulation. In Aim II, we will determine if FoxM1 is required for IL-13/Stat6 signaling to induce goblet cell differentiation in vivo. The IL-13/Stat6 signaling pathway will be activated using intratracheal administration of IL-13 and a Doxycycline-inducible IL-13 transgene. FoxM1 inhibition will be achieved by a genetic approach (IL-13/ CCSP- Cre/ Foxm1-/- mice) and a pharmacological approach (ARF peptide and novel small molecule FoxM1 inhibitors). These experiments will determine whether inactivation of FoxM1 will prevent or decrease IL- 13/Stat6 signaling in airway epithelial cells in vivo. Altogether, these studies will identify molecular mechanisms regulated by FoxM1 in airway epithelial cells and determine the therapeutic benefit of FoxM1 inhibitors in mouse asthma models.

描述（由申请人提供）：与粘液产生增加有关的杯状细胞化生是慢性气道疾病的关键致病特征，例如哮喘，COPD和囊性纤维化，所有这些都导致了全球的大量发病率和死亡率。杯状细胞化生的药理靶向是一个重大的临床挑战。因此，鉴定气道上皮分化中的新分子机制将为治疗慢性气道疾病的治疗提供新的治疗靶标。我们的赠款提案着重于杯状细胞分化中的新分子机制，这些机制由FOXM1调节，FoxM1是Foxm1（Foxhead盒子（FOX）家族的转录因子）。尽管FOXM1在各种癌症的胚胎发育和发病机理中起着重要作用，但FOXM1在慢性气道疾病中的作用尚不清楚。在我们的初步数据中，FOXM1在小鼠的气道上皮细胞中诱导，该细胞具有由OVA，IL-13和House House Dust Mite螨提取物（HDM）引起的哮喘样疾病。 FOXM1在人类哮喘和COPD患者的气道上皮细胞中也被诱导。 FOXM1基因（CCSP-CRE）的遗传缺失或在HDM挑战的气道上皮中FOXM1蛋白（ARF肽）的药理抑制有效地减少了go虫细胞上皮质量，可减少肺部炎症并降低气道对甲基苯乙酸的过度反应性。尽管这些数据表明FOXM1在哮喘发病机理中起关键作用，但由FOXM1调节的分子机制仍然没有表征。我们建议检验以下假设：FOXM1在IL-13/STAT6途径的下游作用，以诱导杯状细胞特异性基因在气道上皮细胞中的表达。在AIM I中，我们将使用FOXM1“功能获得”和“功能丧失”的转基因小鼠在Airway Clara细胞中识别下游FOXM1靶基因对过敏原介导的Clara细胞分化至关重要。此外，我们提供了初步数据，表明IL-13在人类气道上皮细胞和转基因小鼠的气道上皮中诱导FOXM1表达。 FOXM1在体外的敲低抑制了气道上皮细胞响应IL-13刺激的杯状细胞表型的分化。在AIM II中，我们将确定IL-13/STAT6信号传导是否需要FOXM1来诱导杯状细胞分化体内。 IL-13/STAT6信号通路将使用气管内给药IL-13和强力霉素诱导的IL-13转基因激活。 FOXM1抑制作用将通过遗传方法（IL-13/ CCSP- CRE/ FOXM1 - / - 小鼠）和药理学方法（ARF肽和新型小分子FOXM1抑制剂）来实现。这些实验将确定FOXM1的灭活是否会预防或降低体内气道上皮细胞中的IL-13/STAT6信号传导。总的来说，这些研究将确定由FOXM1在气道上皮细胞中调节的分子机制，并确定FOXM1抑制剂在小鼠哮喘模型中的治疗益处。