Deficient Gap Junction Coupling in Two Common Hearing Loss-Related Variants of GJB2.

IF 2.9 3区医学 Q1 OTORHINOLARYNGOLOGY Clinical and Experimental Otorhinolaryngology Pub Date : 2024-08-01 Epub Date: 2024-06-04 DOI:10.21053/ceo.2023.00078

Kaitian Chen, Hongyan Jiang

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Abstract

Objectives: The aim of this study was to explore the functional consequences of two common variants, p.V37I and c.299-300delAT, in the hearing loss-associated gene GJB2.

Methods: Connexin 26 expression and gap junctional permeability were studied in HEK 293T cells transfected with plasmids expressing GJB2 wild-type, p.V37I, or c.299-300delAT CX26 proteins tagged with fluorescent markers. Functional analyses of various GJB2 haplotypes were conducted to thoroughly evaluate alterations in ionic and small-molecule coupling.

Results: The p.V37I protein was localized at the plasma membrane, but it failed to effectively transport intercellular propidium iodide or Ca2+ efficiently, indicating an impairment in both biochemical and ionic coupling. The presence of GJB2 p.V37I seemed to increase the cells' sensitivity to H2O2 treatment. In contrast, the known variant c.299-300delAT protein was not transported to the cell membrane and was unable to form gap junctions, remaining confined to the cytoplasm. Both ionic and biochemical coupling were defective in cells transfected with c.299-300delAT.

Conclusion: The p.V37I and c.299-300delAT GJB2 mutations resulted in deficient gap junction-mediated coupling. Additionally, environmental factors could influence the functional outcomes of the GJB2 p.V37I mutation. These findings could pave the way for the development of molecular therapies targeting GJB2 mutations to treat hearing loss.

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GJB2 中两种常见的听力损失相关变体的间隙连接缺陷。

目的探讨听力损失相关基因 GJB2 的两个常见变体 p.V37I 和 c.299-300delAT 的功能性后果：方法：在转染了表达带有荧光标签的 GJB2 野生型、p.V37I 或 c.299-300delAT CX26 蛋白的质粒的 HEK 293T 细胞中研究了连接蛋白 26 的表达和间隙连接通透性。对不同的 GJB2 单倍型进行了功能分析，以全面评估离子和小分子耦合的改变：结果：p.V37I 蛋白定位于质膜，但不能有效地在细胞间转运碘化丙啶或 Ca2+，这表明生化耦合和离子耦合都受到了影响。GJB2 p.V37I 的存在似乎增加了细胞对 H2O2 处理的敏感性。相反，已知的变体 c.299-300delAT 蛋白不能被转运到细胞膜上，也不能形成间隙连接，而是被限制在细胞质中。c.299-300delAT 转染细胞的离子耦合和生化耦合存在缺陷：结论：p.V37I 和 c.299-300delAT GJB2 突变导致间隙连接介导的耦合缺陷。环境因素可能会影响 GJB2 p.V37I 的功能后果。这些结果可能有助于开发针对 GJB2 突变的听力损失分子疗法。

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来源期刊

Clinical and Experimental Otorhinolaryngology 医学-耳鼻喉科学

CiteScore

4.90

自引率

6.70%

发文量

审稿时长

6-12 weeks

期刊介绍： Clinical and Experimental Otorhinolaryngology (Clin Exp Otorhinolaryngol, CEO) is an international peer-reviewed journal on recent developments in diagnosis and treatment of otorhinolaryngology-head and neck surgery and dedicated to the advancement of patient care in ear, nose, throat, head, and neck disorders. This journal publishes original articles relating to both clinical and basic researches, reviews, and clinical trials, encompassing the whole topics of otorhinolaryngology-head and neck surgery. CEO was first issued in 2008 and this journal is published in English four times (the last day of February, May, August, and November) per year by the Korean Society of Otorhinolaryngology-Head and Neck Surgery. The Journal aims at publishing evidence-based, scientifically written articles from different disciplines of otorhinolaryngology field. The readership contains clinical/basic research into current practice in otorhinolaryngology, audiology, speech pathology, head and neck oncology, plastic and reconstructive surgery. The readers are otolaryngologists, head and neck surgeons and oncologists, audiologists, and speech pathologists.