The epigenetic and genetic profile of KRT1 regulatory elements correlates to phenotypic differences in keratinocyte migration during wound healing

Abstract

The epidermis is a stratified squamous epithelium with each layer of keratinocytes expressing a different set of keratins to reflect the shifting cell states of proliferation and differentiation during growth and repair. This thesis is focused on how keratin 1 (K1), a marker of early differentiation, regulates the rate of migration during wound healing. This was examined by isolating HaCaT clones to represent slow-, moderate- and fast-healing HaCaT cell line, and analyzing if differential KRT1 expression reflected this variation in the rate of wound closure.Bioinformatics was used to identify putative regulatory elements, including enhancers and promoters, in and near the KRT1 locus. The DNA methylation at these putative regulatory elements were assessed using a MSRE-qPCR, to identify if DNA methylation changes correlate with the differential KRT1 expression. Additionally, the identified putative regulatory elements were sequenced using Nanopore sequencing technology, to identify the allelic differences between HaCaT cell lines.This study has provided evidence to suggest KRT1 has a functional role in migration during wound healing, and this is mediated by a dynamic regulatory landscape at the KRT1 locus. The cell model developed in this study can be used to study the migratory role of other keratins during wound healing. Additionally, through bioinformatics analyses, MSRE-qPCR and Nanopore sequencing can be used to assess the epigenetic and genetic profiles of regulatory elements at a given locus.

Date of Award	2019
Original language	English