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The expression profiles of CLDN11-overexpressing and control human induced pluripotent stem cell (iPSC)-derived oligodendrocytes

ABSTRACT: Claudin 11 (CLDN11) is a member of the claudin family. Claudins are cell-cell adhesion molecules situated at tight junctions. The claudin family in mammals comprises 27 four-transmembrane domain proteins. Therein, cerebral CLDN11 is concentrated in oligodendrocytes and its dysregulation has been documented in several CNS disorders. We found that the expression of CLDN11 was significantly upregulated in oligodendrocytes of epileptic patients. To clarify the role of CLDN11 in oligodendrocytes in epileptic pathology, we transfected the human iPSC-derived oligodendrocytes with CLDN11 plasmid or vector control plasmid for 48 hours, respectively. Subsequently, three CLDN11-overexpressing and three vector control human iPSC-derived oligodendrocyte samples were subject to RNA sequencing.

ORGANISM(S): Homo sapiens

PROVIDER: GSE274180 | GEO | 2024/08/10

REPOSITORIES: GEO

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Project description:Claudins are tight junction proteins that are involved in tight junction formation and function. Previous studies have shown that claudin-7 is frequently upregulated in epithelial ovarian cancer (EOC) along with claudin-3 and claudin-4. Here, we investigate in detail the expression patterns of claudin-7, as well as its possible functions in EOC. Initially a total of 95 ovarian tissue samples (7 normal ovarian tissues, 65 serous carcinomas, 11 clear cell carcinomas, 8 endometrioid carcinomas and 4 mucinous carcinomas) were studied for claudin-7 expression using RT-PCR analysis. The gene for claudin-7, CLDN7, was found to be significantly upregulated in all the tumor tissue samples studied. Similarly, immunohistochemical analysis and western blotting showed that claudin-7 protein was significantly overexpressed in the vast majority of EOCs. Two cell lines(OVCAR-2 and OVCA420) were chosen for microarray based gene expression analysis using small interfering RNA-mediated(SiRNA) knockdown of claudin-7. This treatment led to significant changes in gene expression that was validated by RT-PCR and immunoblotting. Ovarian cancer lines OVCA420 and OVCAR-2 were cultured in duplicate in McCoy’s 5A growth medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS) and antibiotics (100 units/ml penicillin and 100 mg/ml streptomycin). Cells cultured in 6-well plates were transfected with either Claudin-7 specific siRNA oligos duplexes (Ambion, Inc.Austin, TX) or control siRNA #1 (Dharmacon) using LipofectAMINE 2000 (Invitrogen). Cells were treated for 72 hours to allow maximum knockdown, after which they were harvested for RNA preparation. Total RNA, was extracted using Trizol, quality and quantity were checked using the Agilent 2100 Bioanalyzer with RNA 6000 Nano chip, (Agilent Technologies UK Ltd, West Lothian, UK). Biotinylated cRNA was generated using the Illumina TotalPrep RNA Amplification Kit (Ambion; Austin, TX) starting with approximately 500 ng total RNA. Hybridization of the cRNA was to the Sentrix HumanRef-8 Expression BeadChip (Illumina, Inc., San Diego, CA). Microarray data processing and analysis was performed using Illumina Bead Studio software. Hierarchical clustering analysis of the significant genes was done using the JMP 6.0.0 software. Validation of the expression patterns of selected genes was performed using RT-PCR.

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The expression profiles of CLDN11-overexpressing and control human induced pluripotent stem cell (iPSC)-derived oligodendrocytes

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