WS14.06Novel targets to regulate the abundance of G542X CFTR mRNA

IF 5.4 2区医学 Q1 RESPIRATORY SYSTEM

Journal of Cystic Fibrosis Pub Date : 2025-06-01 DOI:10.1016/j.jcf.2025.03.575

A.F. Henriques, A. Abrantes, C.M. Farinha

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引用次数: 0

Abstract

Background and Objectives

The CFTR gene contains over 2,100 documented variants classified into 7 classes based on their impact on CFTR protein. Despite the advances with the modulators, 10-15% of patients, including those with nonsense variants (Class I), lack targeted therapies. Nonsense variants, such as the prevalent G542X, introduce premature stop codons (PTCs), triggering nonsense-mediated decay (NMD) and degrading CFTR mRNA. Personalized therapies for these variants remain an unmet need.

Methods and Results

As we did before for common variants (1), we focused on transcriptomic and proteomic profiles of 16HBE14o- cells bearing G542X-CFTR. Differential expression analysis revealed 201 transcripts, and 35 proteins uniquely altered in G542X cells. Enrichment analysis highlighted processes like “regulation of transcription,” “RNA metabolic process,” and “RNA-binding,” suggesting key roles in CFTR mRNA stability.

Validation identified 7 upregulated and 4 downregulated genes. Additionally, we found that the knockdown of the RNA binding proteins TUT1 (Terminal Uridylyl Transferase 1) and HNRNPUL2 (Heterogeneous Nuclear Ribonucleoprotein U Like 2) has a considerable effect in the levels of CFTR mRNA in G542X cells. A similar effect was found under overexpression of ZNF793 (Zinc Finger Protein 793) in the same G542X cell line. Ongoing work aims to assess protein expression and explore combinatory strategies for synergistic therapeutic outcomes.

Conclusion

We identified novel RNA-binding proteins as regulators of CFTR mRNA abundance in G542X cells, paving the way for targeted therapies combining NMD inhibition and PTC readthrough to restore CFTR expression in patients lacking current treatment options.

Acknowledgements: Work supported by Emily's Entourage and center grants to BioISI UIDB/04046/2020 and UIDP/04046/2020 (https://doi. org/10.54499/UIDB/04046/2020) from FCT, Portugal (to BioISI).

[1] www.cftr2.org

[2] Santos L et al (2023). Cell Biosci 13, 26

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ws14.06调节G542X CFTR mRNA丰度的新靶点

背景与目的CFTR基因包含超过2100个已记录的变异，根据其对CFTR蛋白的影响将其分为7类。尽管调节剂取得了进展，但仍有10-15%的患者，包括无义变异（I类）患者，缺乏靶向治疗。无义变异，如普遍存在的G542X，引入过早终止密码子（ptc），触发无义介导的衰变（NMD）并降解CFTR mRNA。针对这些变异的个性化治疗仍然是一个未满足的需求。方法和结果与之前对常见变异(1)的研究一样，我们重点研究了携带G542X-CFTR的16HBE14o-细胞的转录组学和蛋白质组学特征。差异表达分析显示，201个转录本和35个蛋白在G542X细胞中有独特的改变。富集分析强调了“转录调节”、“RNA代谢过程”和“RNA结合”等过程，表明CFTR mRNA稳定性的关键作用。验证鉴定出7个上调基因和4个下调基因。此外，我们发现RNA结合蛋白TUT1（终端尿酰基转移酶1）和HNRNPUL2（异质核核糖核蛋白U样2）的敲低对G542X细胞中CFTR mRNA的水平有相当大的影响。在相同的G542X细胞系中，过表达ZNF793（锌指蛋白793）也有类似的效果。正在进行的工作旨在评估蛋白质表达并探索协同治疗结果的组合策略。结论：我们发现了新的rna结合蛋白作为G542X细胞中CFTR mRNA丰度的调节因子，为NMD抑制和PTC读透联合靶向治疗在缺乏当前治疗方案的患者中恢复CFTR表达铺平了道路。感谢：Emily’s Entourage支持的工作，以及BioISI UIDB/04046/2020和UIDP/04046/2020的中心资助（https://doi）。/10.54499/ uidb /04046/2020)从葡萄牙FCT（到BioISI）。[1] www.cftr2.org[2] Santos等（2023）。细胞生物学报13,26

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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.