[09.06]囊性纤维化上皮细胞中HE4表达与上皮间质转化关系的研究

IF 5.4 2区医学 Q1 RESPIRATORY SYSTEM

Journal of Cystic Fibrosis Pub Date : 2025-06-01 DOI:10.1016/j.jcf.2025.03.545

M. Pócsi , G.J. Balla , F. Fenyvesi , Á. Rusznyák , Z. Fejes , I. Balogh , M. Macek Jr. , M.D. Amaral , B. Nagy Jr.

{"title":"[09.06]囊性纤维化上皮细胞中HE4表达与上皮间质转化关系的研究","authors":"M. Pócsi , G.J. Balla , F. Fenyvesi , Á. Rusznyák , Z. Fejes , I. Balogh , M. Macek Jr. , M.D. Amaral , B. Nagy Jr.","doi":"10.1016/j.jcf.2025.03.545","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><div>Elevated expression of human epididymis protein 4 (HE4) was described in cystic fibrosis (CF), which was directly influenced by impaired CFTR via NF-κB in p.Phe508del-CFTR CFBE 41o− cells <em>in vitro</em>. Dysfunctional CFTR was also associated with epithelial mesenchymal transition (EMT) in CF. However, no data is available if HE4 expression alters and is linked to CF-related EMT.</div></div><div><h3>Methods</h3><div>EMT characteristics were compared between CFBE 41o− cells expressing p.Phe508del-CFTR and wt-CFTR through the expression of epithelial and mesenchymal markers by RT-qPCR and fluorescence microscopy. EMT was further triggered in p.Phe508del-CFTR CFBE cells by TGF-β1 from 24-96h to investigate the relationship of EMT with HE4 and MMP9 expression. To restore CFTR dysfunction, VX-445/VX-661/VX-770 was applied to monitor EMT phenotype. The direct effect of abnormal HE4 expression on EMT and MMP9 was analyzed when: <em>i)</em> mutant CFBE cells were treated with recombinant HE4, and <em>ii)</em> HE4 expression was artificially reduced by transfection with HE4 specific siRNA.</div></div><div><h3>Results</h3><div>Untreated p.Phe508del-CFTR CFBE cells had a trend to be more mesenchymal than wt-CFTR CFBE cells and showed higher HE4 and MMP9 levels. However, in response to TGF-β1, E-cadherin levels were lower, while those of N-cadherin and MMP9 were higher in p.Phe508del-CFTR CFBE cells. In parallel, HE4 levels were decreased already after 48h and further reduced until 96h. The EMT phenotype could be reversed by CFTR modulators causing a decrease in both HE4 and MMP9 levels. Finally, both treatment with recombinant HE4 and downregulated HE4 levels promoted EMT with induced MMP9 in p.Phe508del-CFTR CFBE cells.</div></div><div><h3>Conclusion</h3><div>High baseline HE4 levels in CF may contribute to the development of EMT accompanied with decreasing intracellular HE4 and upregulated MMP9 expression in airway epithelial cells.</div><div><strong>Grants:</strong> This study is supported by FK-135327 grant (BN) and UID/04046/2025 centre grant in MDA lab (to BioISI) from FCT/MCTES Portugal.</div></div>","PeriodicalId":15452,"journal":{"name":"Journal of Cystic Fibrosis","volume":"24 ","pages":"Page S19"},"PeriodicalIF":5.4000,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"WS09.06Investigation of HE4 expression in relation to epithelial mesenchymal transition in cystic fibrosis epithelial cells\",\"authors\":\"M. Pócsi , G.J. Balla , F. Fenyvesi , Á. Rusznyák , Z. Fejes , I. Balogh , M. Macek Jr. , M.D. Amaral , B. Nagy Jr.\",\"doi\":\"10.1016/j.jcf.2025.03.545\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><div>Elevated expression of human epididymis protein 4 (HE4) was described in cystic fibrosis (CF), which was directly influenced by impaired CFTR via NF-κB in p.Phe508del-CFTR CFBE 41o− cells <em>in vitro</em>. Dysfunctional CFTR was also associated with epithelial mesenchymal transition (EMT) in CF. However, no data is available if HE4 expression alters and is linked to CF-related EMT.</div></div><div><h3>Methods</h3><div>EMT characteristics were compared between CFBE 41o− cells expressing p.Phe508del-CFTR and wt-CFTR through the expression of epithelial and mesenchymal markers by RT-qPCR and fluorescence microscopy. EMT was further triggered in p.Phe508del-CFTR CFBE cells by TGF-β1 from 24-96h to investigate the relationship of EMT with HE4 and MMP9 expression. To restore CFTR dysfunction, VX-445/VX-661/VX-770 was applied to monitor EMT phenotype. The direct effect of abnormal HE4 expression on EMT and MMP9 was analyzed when: <em>i)</em> mutant CFBE cells were treated with recombinant HE4, and <em>ii)</em> HE4 expression was artificially reduced by transfection with HE4 specific siRNA.</div></div><div><h3>Results</h3><div>Untreated p.Phe508del-CFTR CFBE cells had a trend to be more mesenchymal than wt-CFTR CFBE cells and showed higher HE4 and MMP9 levels. However, in response to TGF-β1, E-cadherin levels were lower, while those of N-cadherin and MMP9 were higher in p.Phe508del-CFTR CFBE cells. In parallel, HE4 levels were decreased already after 48h and further reduced until 96h. The EMT phenotype could be reversed by CFTR modulators causing a decrease in both HE4 and MMP9 levels. Finally, both treatment with recombinant HE4 and downregulated HE4 levels promoted EMT with induced MMP9 in p.Phe508del-CFTR CFBE cells.</div></div><div><h3>Conclusion</h3><div>High baseline HE4 levels in CF may contribute to the development of EMT accompanied with decreasing intracellular HE4 and upregulated MMP9 expression in airway epithelial cells.</div><div><strong>Grants:</strong> This study is supported by FK-135327 grant (BN) and UID/04046/2025 centre grant in MDA lab (to BioISI) from FCT/MCTES Portugal.</div></div>\",\"PeriodicalId\":15452,\"journal\":{\"name\":\"Journal of Cystic Fibrosis\",\"volume\":\"24 \",\"pages\":\"Page S19\"},\"PeriodicalIF\":5.4000,\"publicationDate\":\"2025-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Cystic Fibrosis\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1569199325006411\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"RESPIRATORY SYSTEM\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Cystic Fibrosis","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1569199325006411","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"RESPIRATORY SYSTEM","Score":null,"Total":0}

引用次数: 0

摘要

目的探讨人附睾蛋白4 （HE4）在囊性纤维化（CF）中表达的升高，并通过NF-κB直接影响pph508del -CFTR CFBE 410−细胞中CFTR受损。CFTR功能障碍也与CF的上皮间充质转化（EMT）相关。然而，HE4表达改变是否与CF相关的EMT相关尚无数据。方法采用RT-qPCR和荧光显微镜对表达p.Phe508del-CFTR和wt-CFTR的CFBE 410−细胞的上皮和间质标记物的semt特征进行比较。在p.Phe508del-CFTR CFBE细胞中，通过TGF-β1在24-96h进一步触发EMT，研究EMT与HE4和MMP9表达的关系。为了恢复CFTR功能障碍，应用VX-445/VX-661/VX-770监测EMT表型。分析HE4异常表达对EMT和MMP9的直接影响：1)用重组HE4处理突变体CFBE细胞，2)通过转染HE4特异性siRNA人工降低HE4表达。结果经处理的p.Phe508del-CFTR CFBE细胞比wt-CFTR CFBE细胞间质化趋势明显，HE4和MMP9水平较高。而在p.Phe508del-CFTR CFBE细胞中，受TGF-β1的影响，E-cadherin水平较低，N-cadherin和MMP9水平较高。同时，HE4水平在48h后已经降低，并进一步降低至96h。CFTR调节剂可以逆转EMT表型，导致HE4和MMP9水平降低。最后，重组HE4和下调HE4水平均可促进p.Phe508del-CFTR CFBE细胞中诱导MMP9的EMT。结论CF高基线HE4水平可能与EMT的发生有关，同时伴有气道上皮细胞HE4降低和MMP9表达上调。本研究由FCT/MCTES葡萄牙MDA实验室（BioISI）的FK-135327拨款（BN）和UID/04046/2025中心拨款支持。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

WS09.06Investigation of HE4 expression in relation to epithelial mesenchymal transition in cystic fibrosis epithelial cells

Objective

Elevated expression of human epididymis protein 4 (HE4) was described in cystic fibrosis (CF), which was directly influenced by impaired CFTR via NF-κB in p.Phe508del-CFTR CFBE 41o− cells in vitro. Dysfunctional CFTR was also associated with epithelial mesenchymal transition (EMT) in CF. However, no data is available if HE4 expression alters and is linked to CF-related EMT.

Methods

EMT characteristics were compared between CFBE 41o− cells expressing p.Phe508del-CFTR and wt-CFTR through the expression of epithelial and mesenchymal markers by RT-qPCR and fluorescence microscopy. EMT was further triggered in p.Phe508del-CFTR CFBE cells by TGF-β1 from 24-96h to investigate the relationship of EMT with HE4 and MMP9 expression. To restore CFTR dysfunction, VX-445/VX-661/VX-770 was applied to monitor EMT phenotype. The direct effect of abnormal HE4 expression on EMT and MMP9 was analyzed when: i) mutant CFBE cells were treated with recombinant HE4, and ii) HE4 expression was artificially reduced by transfection with HE4 specific siRNA.

Results

Untreated p.Phe508del-CFTR CFBE cells had a trend to be more mesenchymal than wt-CFTR CFBE cells and showed higher HE4 and MMP9 levels. However, in response to TGF-β1, E-cadherin levels were lower, while those of N-cadherin and MMP9 were higher in p.Phe508del-CFTR CFBE cells. In parallel, HE4 levels were decreased already after 48h and further reduced until 96h. The EMT phenotype could be reversed by CFTR modulators causing a decrease in both HE4 and MMP9 levels. Finally, both treatment with recombinant HE4 and downregulated HE4 levels promoted EMT with induced MMP9 in p.Phe508del-CFTR CFBE cells.

Conclusion

High baseline HE4 levels in CF may contribute to the development of EMT accompanied with decreasing intracellular HE4 and upregulated MMP9 expression in airway epithelial cells.

Grants: This study is supported by FK-135327 grant (BN) and UID/04046/2025 centre grant in MDA lab (to BioISI) from FCT/MCTES Portugal.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Journal of Cystic Fibrosis 医学-呼吸系统

CiteScore

10.10

自引率

13.50%

发文量

1361

审稿时长

50 days

期刊介绍： The Journal of Cystic Fibrosis is the official journal of the European Cystic Fibrosis Society. The journal is devoted to promoting the research and treatment of cystic fibrosis. To this end the journal publishes original scientific articles, editorials, case reports, short communications and other information relevant to cystic fibrosis. The journal also publishes news and articles concerning the activities and policies of the ECFS as well as those of other societies related the ECFS.