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A Single-Cell Transcriptomic Atlas of Human Lens Homeostasis and Aging

ABSTRACT: Lens epithelial cells (LECs) are crucial for lens transparency and function. Aging disrupts LEC homeostasis and contributes to the development of age-related cataracts (ARCs). The lens epithelium may rely on adult stem/progenitor cells to sustain homeostasis and counteract aging-related factors. While the existence of stem cells in the lens epithelium has been supported by various studies, identifying these cells remains challenging. Recent advancements in single-cell RNA sequencing (scRNA-seq) provide high-resolution insights into cellular heterogeneity and offer a promising approach to elucidate the presence, origin, and age-related changes of lens stem/progenitor cells. We collected human donor lenses with ethical approval for scRNA-seq analysis, including samples from four non-aged (<65 years) and four aged (>65 years) individuals. After quality assessment and filtering, the transcriptome profiles of a total of 56,717 cells from two groups were depicted and merged to allow identification of overlapping cell populations in the following analysis. Subsequent analyses involved cell (sub)type characterization, trajectory inference, and cell-cell communication. Six types of lens superficial cells were identified by scRNA-seq, including four categories of LECs: early differentiation state LECs (eLECs), middle differentiation state LECs (mLECs), late differentiation state LECs (lLECs), and transient amplifying cells (TACs), along with a small number of lens fibre cells and immune cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE287919 | GEO | 2025/05/01

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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A Single-Cell Transcriptomic Atlas of Human Lens Homeostasis and Aging

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