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Transcriptome, proteome, and metabolome of lens fiber cells from mice exposed to cigarette smoke

ABSTRACT: We adopted an omics (transcriptome, proteome, and metabolome) approach to characterize the differentially regulated molecules in mouse lens fiber cells exposed to cigarette smoke (CS). Eight pregnant female mice (gestation days 19-20) were placed in a smoking chamber that served as a CS-exposed group. The mothers and newborn pups were exposed to CS for five hours/day, five days/week for 110 days. In parallel, mice were kept in normal cages that served as a control group. The ocular lenses were isolated and fiber cells were separated from the lens epithelium under a microscope. The control and CS-exposed groups were maintained in four biological replicates each consisting of a pool of lens fiber cells from four eyes. The eight biological replicates (four control and four CS-exposed) of fiber cells were used for the next-generation-based transcriptome (RNA-Seq) sequencing, mass-spectrometry-based proteome, and metabolome profiling. RNA-Seq analysis identified the expression (≥1.0 FPKM) of 9,590 and 9,531 genes in control and CS-exposed fiber cells, respectively. The analysis identified 348 differentially expressed genes, which included 186 downregulated and 162 upregulated genes in CS-exposed fiber cells. Proteome profiling revealed a total of 2,424 proteins in control and CS-exposed fiber cells. Metabolome profiling identified a total of 280 metabolites, marked with decreased levels of branched-chain amino acids (BCAAs)-related metabolites in CS-exposed fiber cells. In conclusion, we have established a comprehensive omics profile of CS-exposed lens fiber cells. To the best of our knowledge, this is the first report investigating a comprehensive omics profile of CS-exposed fiber cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE144818 | GEO | 2021/09/29

REPOSITORIES: GEO

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Project description:The mature eye lens contains a surface layer of epithelial cells called the lens epithelium that require a functional mitochondrial population to maintain the homeostasis and transparency of the entire lens. The lens epithelium overlies a core of terminally differentiated fiber cells that must degrade their mitochondria to achieve lens transparency. These distinct mitochondrial populations make the lens a useful model system to identify those genes that regulate the balance between mitochondrial homeostasis and elimination. Here we used an RNA sequencing and bioinformatics approach to identify the transcript levels of all genes expressed by distinct regions of the lens epithelium and maturing fiber cells of the embryonic Gallus gallus (chicken) lens. Our analysis detected over 15,000 unique transcripts expressed by the embryonic chicken lens. Of these, over 3000 transcripts exhibited significant differences in expression between lens epithelial cells and fiber cells. Multiple transcripts coding for separate mitochondrial homeostatic and degradation mechanisms were identified to exhibit preferred patterns of expression in lens epithelial cells that require mitochondria relative to lens fiber cells that require mitochondrial elimination. These included differences in the expression levels of metabolic, autophagy, and mitophagy transcripts between lens epithelial cells and lens fiber cells. These data provide a comprehensive window into all genes transcribed by the lens and those mitochondrial regulatory and degradation pathways that function to maintain mitochondrial populations in the lens epithelium and to eliminate mitochondria in maturing lens fiber cells. Differentiation-state transcriptional analysis of embryonic chicken lenses was performed following microdissection of 100 embryonic day 13 (E13) chicken lenses into four distinct regions that represent a continuum of lens cell differentiation states: lens central epithelium (EC), equatorial epithelium (EQ), cortical fibers (FP), and central fibers (FC). Further analysis of the transcriptional content of biologically replicate samples was performed by Illumina directional mRNA sequencing and resulting reads mapped by TopHat and assembled with Cufflinks.

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Transcriptome, proteome, and metabolome of lens fiber cells from mice exposed to cigarette smoke

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