Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A Keratin 12 Expression-Based Analysis of Stem-Precursor Cells and Differentiation in The Limbal-Corneal Epithelium Using Single-Cell RNA-Seq Data

Version 1 : Received: 18 January 2024 / Approved: 18 January 2024 / Online: 18 January 2024 (15:54:32 CET)

A peer-reviewed article of this Preprint also exists.

Wolosin, J.M. A Keratin 12 Expression-Based Analysis of Stem-Precursor Cells and Differentiation in the Limbal–Corneal Epithelium Using Single-Cell RNA-Seq Data. Biology 2024, 13, 145. Wolosin, J.M. A Keratin 12 Expression-Based Analysis of Stem-Precursor Cells and Differentiation in the Limbal–Corneal Epithelium Using Single-Cell RNA-Seq Data. Biology 2024, 13, 145.

Abstract

The corneal epithelium (CE) is spread between two domains, the outer vascularized limbus and the avascular cornea proper. Epithelial cells undergo constant migration from the limbus to the vision-critical central cornea. Coordinated with this migration the cells undergo differentiation changes where a pool of unique stem/precursor cells at the limbus yields the mature cells that reach the corneal center. Differentiation is heralded by the expression of the corneal-specific Krt12. Processing data acquired by scRNA-Seq we found that the increase of Krt12 expression occurs in four distinct steps within the limbus plus a single continuous increase in the cornea. Differential gene analysis demonstrated that these domains reflect discreet stages of CE differentiation and yielded extensive information of the genes undergoing down or up regulation in the sequential transition from less to more differentiate conditions. The approach allowed the identification of multiple gene cohorts, including a) the genes that have maximal expression in the most primitive, Krt12-negative cell cohort, which is likely to include the stem/precursor cells; b) the sets of genes that undergo continuous increase or decrease along the whole differentiation path, and c) and the genes showing maximal positive or negative correlation with the changes in Krt12.

Keywords

scRNA-Seq, cornea, limbus, epithelium, Krt12, stem cells, differentiation, gene expression

Subject

Biology and Life Sciences, Cell and Developmental Biology

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