FAM134B-induced endoplasmic reticulum (ER)-phagy exacerbates cisplatin-insulted hair cell apoptosis ：Possible relation to excessive ER stress

Huiming Yang; Haiyan Yin; Yue Wang; Jisheng Liu; Lingchuan Guo; Hao Zhao; Xiaohui Bai; Jianfeng Li; Qianqian Yang

doi:10.1016/j.abb.2023.109766

FAM134B-induced endoplasmic reticulum (ER)-phagy exacerbates cisplatin-insulted hair cell apoptosis ：Possible relation to excessive ER stress

Arch Biochem Biophys. 2023 Oct 15:748:109766. doi: 10.1016/j.abb.2023.109766. Epub 2023 Oct 9.

Authors

Huiming Yang¹, Haiyan Yin², Yue Wang³, Jisheng Liu⁴, Lingchuan Guo⁵, Hao Zhao¹, Xiaohui Bai⁶, Jianfeng Li⁷, Qianqian Yang⁸

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, 250012, China.
² School of Basic Medical Science, Jining Medical University, Jining, Shandong, China.
³ Department of Otolaryngology Head and Neck Surgery, Ningbo First Hospital, Ningbo, Zhejiang, 315000, China.
⁴ Department of Otolaryngology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215123, China.
⁵ Department of Pathology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215123, China.
⁶ Institute of Eye and ENT, Provincial Hospital affiliated to Shandong First Medical University, Jinan, Shandong, 250021, China.
⁷ Department of Otolaryngology-Head and Neck Surgery, Shandong Provincial Hospital, Shandong University, Jinan, Shandong, 250012, China; Institute of Eye and ENT, Provincial Hospital affiliated to Shandong First Medical University, Jinan, Shandong, 250021, China. Electronic address: LiJF@email.sdu.edu.cn.
⁸ Department of Pathology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, 215123, China. Electronic address: yangqianqian@suda.edu.cn.

PMID: 37813237
DOI: 10.1016/j.abb.2023.109766

Abstract

Aims: FAM134B, the initial endoplasmic reticulum (ER)-phagy receptor identified, facilitates ER-phagy during ER stress. The malfunction of FAM134B has been demonstrated to have a crucial role in the pathological mechanisms of diverse human ailments. However, the role of FAM134B-mediated ER-phagy in ototoxicity, particularly in cisplatin-induced ototoxicity, remains unclear. The present study endeavors to investigate whether FAM134B is expressed in House Ear Institute-Organ of Corti 1 (HEI-OC1) and C57BL/6 murine cochlear hair cells (HCs), and to explore its potential function in cisplatin-mediated ototoxicity, with the aim of discovering new insights that can mitigate or forestall the irreversible adverse effect of cisplatin.

Methods: Immunofluorescence (IF) staining was used to test the expression pattern of FAM134B, levels of C/EBP-homologous protein (CHOP), autophagy, and co-localization ratio of lysosomes and ER. Western blotting was employed to measure changes in expression levels of FAM134B, LC3B, ER stress-related proteins, LAMP1 and apoptotic mediators. Cell apoptosis was examined using transferase dUTP nick end labeling (TUNEL) assay and flow cytometry.

Results: In the present investigation, it was observed that FAM134B exhibited a diffuse expression pattern in the cytoplasm and nuclei of control HEI-OC1 cells. Following cisplatin administration, FAM134B was found to accumulate and form distinct dots around the nuclei, concomitant with increased levels of ER-phagy, ER stress, unfolded protein response (UPR), and cell apoptosis. Additionally, knockdown of FAM134B resulted in reduced ER-phagy, mitigated ER stress and UPR, and decreased apoptotic activity in HEI-OC1 cells following cisplatin exposure.

Conclusions: Collectively, the findings of this study demonstrate that FAM134B-mediated ER-phagy enhances the susceptibility of HCs to ER stress and apoptosis in response to cisplatin-induced stress. This suggests a sequential progression of ER-phagy, ER stress and apoptosis following cisplatin stimulus, and implies the potential therapeutic benefit of inhibiting of FAM134B-mediated ER-phagy in the prevention of cisplatin-related ototoxicity.

Keywords: Apoptosis; Cisplatin; ER-Phagy; FAM134B; HEI-OC1 cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis
Autophagy
Cisplatin* / toxicity
Endoplasmic Reticulum / metabolism
Endoplasmic Reticulum Stress
Hair Cells, Auditory / metabolism
Humans
Mice
Ototoxicity* / metabolism

Substances

Cisplatin