Forcing keratinocyte stem cells out of their niche
If you have any partnering opportunities for Regener8 members, please contact us.
NESCI would be interested in funding / studentship opportunities for the following proposal.
NESCI are interested in industrial partners who may like to collaborate in one of these fields (for a CASE studentship or other awards), or in anyone who is interested in these areas of research (including other academic groups). For further details, please contact Helen Clamp from NESCI.
Project: Forcing keratinocyte stem cells out of their niche
Lead: Dr Julia Reichelt
Cells and tissues are able to respond to physical force. Mechanical stress can cause diverse biological responses like cell proliferation or differentiation and is involved in developmental processes and regeneration.
In the epidermis keratinocytes are exposed to a wide range of loads. Multipotent keratinocyte stem cells (KSC) residing in the hair follicle bulge are unique being additionally subject to tensile forces from the arrector pili muscle (APM) which inserts at the bulge region. The connection between KSC niche and this muscle suggests a specific mechanical regulation of KSC that will be investigated in this PhD project.
Wnt and β-catenin are known to be essential for hair follicle development, cycling and de novo hair follicle growth. The project will focus on studying the involvement of Wnt pathways in mechanical signalling. KSC spheres, the generation of which has been recently established in this lab and which mimic the bulge environment, will be biaxially stretched to different parameters using the Flexcell system. The Flexcell apparatus is a computer-controlled vacuum system allowing stretching cells grown on flexible supports. The cellular response will be studied using Western blotting, immunofluorescence analysis, TCF reporter plasmid (TOPflash) assays, qPCR and life cell imaging. Results obtained from cell culture experiments will be transferred to wound healing studies including pharmacological stimulators/inhibitors of the APM in transgenic KSC reporter mice (Krt15CrePR;R26R) which allow tracking of KSC and their progeny (HO licence in place). Migration, proliferation and differentiation of labelled KSC will be analysed on skin sections and in whole mount preparations, using BrdU labelling and keratinocyte differentiation markers respectively. Clonal analysis of keratinocytes isolated from wounds will reveal whether APM stimulation may result in depletion of KSC from their niche.
The ultimate goal of this research is to establish whether epidermal KSC can be stimulated to increase skin wound healing through manipulation of the APM via pharmacological agents.
