DOI: 10.5061/dryad.b2rbnzsrj
title: GSDMD-mediated mitochondrial dysfunction in marginal cells: A potential driver of inflammation and stria vascularis damage in CIHL

description: Inflammation is among the known causes of cisplatin-induced hearing loss
 (CIHL), but its exact pathophysiological mechanisms remain unclear.
 Herein, we demonstrated that pyroptosis—a recently identified inflammatory
 type of regulated cell death dependent on gasdermin D (GSDMD)—was
 activated in the cochleae of cisplatin-treated mice, causing CIHL.
 Meanwhile, treatment with the GSDMD inhibitor necrosulfonamide alleviated
 CIHL in these mice. To further examine the role of GSDMD-mediated
 pyroptosis in CIHL, we conducted experiments in Gsdmd-deficient
 mice. Gsdmd−/− mice demonstrated significantly lower cisplatin-induced
 cochlear damage than control mice and appeared to be invulnerable to CIHL.
 Furthermore, GSDMD-mediated pyroptosis in the stria vascularis (SV), but
 not in the hair cells (HCs), played a dominant role in CIHL. In marginal
 cells (MCs) of SV, cisplatin induced caspase-dependent GSDMD cleavage, and
 the pore-forming N-terminal of GSDMD rapidly localized to the
 mitochondria, leading to abnormal mitochondrial aggregation and oxidative
 stress. The consequent mitochondrial dysfunction in MCs might result in
 the severe progression of inflammation, SV damage, and HC loss. Notably,
 the pharmacological inhibition of pyroptosis using the FDA-approved drug
 disulfiram effectively alleviated the symptoms of CIHL. Collectively,
 these findings offer a broad avenue for inhibiting pyroptosis-induced
 cisplatin ototoxicity and provide valuable theoretical insights for the
 clinical management of CIHL.
description: # GSDMD-mediated mitochondrial dysfunction in marginal cells: A potential
 driver of inflammation and stria vascularis damage in CIHL
 [https://doi.org/10.5061/dryad.b2rbnzsrj
 ](https://doi.org/10.5061/dryad.b2rbnzsrj) ## Description of the data and
 file structure  To investigate the role of GSDMD in cisplatin-induced
 ototoxicity, we performed transcriptomic analysis on the cochleae of four
 experimental groups: (1) wild-type (WT) mice, (2) WT mice treated with
 cisplatin (WT+cisplatin), (3) *Gsdmd *knockout mice (*Gsdmd−/−*), and (4)
 *Gsdmd−/−* mice treated with cisplatin (*Gsdmd−/−*+cisplatin). Total RNA
 was extracted from bilateral cochleae (3 mice per group) using the TRIzol
 reagent (Vazyme, R411-01). The extracted RNA was sent to Igenecode Gene
 Technology Corporation (Beijing) for library establishment and sequencing
 analysis. Briefly, the constructed libraries were prepared and quantified
 using the Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR
 System and sequenced on the Illumina sequencing platform. Raw data
 obtained from sequencing were filtered, and the clean reads obtained were
 aligned to the reference genome. Differentially expressed genes (DEGs)
 were identified using the DESeq2 package with a threshold of |log2 fold
 change| &gt; 1 and an adjusted *p*-value &lt; 0.05. ### Files and
 variables #### File: All\_GeneDiffExp.xls **Description:** The data is
 the  transcriptomic analysis on the cochleae of four experimental groups:
 (1) wild-type (WT) mice, (2) WT mice treated with cisplatin
 (WT+cisplatin), (3) *Gsdmd* knockout mice (*Gsdmd−/−*), and (4) *Gsdmd−/−*
 mice treated with cisplatin (*Gsdmd−/−*+cisplatin). In this table,
 "WT" refers to WT, "WT_with_Cis" refers
 to WT+cisplatin, "KO" refers to *Gsdmd−/−*,
 "KO_with_Cis" refers to *Gsdmd−/−*+cisplatin. In the document,
 each variable (column name) and the corresponding explanation are shown
 below: (1) GeneID: The gene Entrez ID (2)
 log2FoldChange(KO-VS-KO_with_Cis.DEseq2): log2-transformed fold change in
 gene expression between KO and KO_with_Cis . (3)
 Padj(KO-VS-KO_with_Cis.DEseq2): Adjusted p-value of the fold change (KO
 vs. KO_with_Cis). (4) Up/Down-Regulation(KO_with_Cis/KO): Direction of
 regulation: "Up" if expression is higher in KO_with_Cis vs. KO,
 "Down" if lower. (5) log2FoldChange(WT-VS-KO.DEseq2):
 log2-transformed fold change in gene expression between WT and KO . (6)
 Padj(WT-VS-KO.DEseq2): Adjusted p-value of the fold change (WT vs. KO).
 (7) Up/Down-Regulation(KO/WT): Direction of regulation: "Up" if
 expression is higher in KO vs. WT, "Down" if lower. (8)
 log2FoldChange(WT-VS-KO_with_Cis.DEseq2): log2-transformed fold change in
 gene expression between WT and KO_with_Cis. (9)
 Padj(WT-VS-KO_with_Cis.DEseq2): Adjusted p-value of the fold change (WT
 vs. KO_with_Cis) (10) Up/Down-Regulation(KO_with_Cis/WT): Direction of
 regulation: "Up" if expression is higher in KO_with_Cis vs. WT,
 "Down" if lower. (11) log2FoldChange(WT-VS-WT_with_Cis.DEseq2):
 log2-transformed fold change in gene expression between WT 
 and WT_with_Cis. (12) Padj(WT-VS-WT_with_Cis.DEseq2): Adjusted p-value of
 the fold change (WT vs. WT_with_Cis) (13)
 Up/Down-Regulation(WT_with_Cis/WT): Direction of regulation:
 "Up" if expression is higher in WT_with_Cis vs. WT,
 "Down" if lower. (14)
 log2FoldChange(WT_with_Cis-VS-KO_with_Cis.DEseq2): log2-transformed fold
 change in gene expression between WT_with_Cis and KO_with_Cis. (15)
 Padj(WT_with_Cis-VS-KO_with_Cis.DEseq2): Adjusted p-value of the fold
 change (WT_with_Cis vs. KO_with_Cis) (16)
 Up/Down-Regulation(KO_with_Cis/WT_with_Cis): Direction of regulation:
 "Up" if expression is higher in KO_with_Cis vs. WT_with_Cis,
 "Down" if lower. (17) Symbol: Gene symbol (18) Description:
 Functional description of the gene. (19) GeneType: Classification of the
 gene type. Additionally, cells marked with * indicate: Non-significant
 results (Padj &gt; 0.05), low-count genes excluded by DESeq2, or data
 filtered during quality control. ##### Variables The dataset includes two
 variables: (1) cisplatin treatment (with or without cisplatin) and (2)
 *Gsdmd* gene status (wild-type or knockout).