Topic status: We're looking for students to study this topic.
Micro RNA’s have recently emerged as key regulators of gene expression during many cellular processes, especially mammalian cell growth and development. Recent evidence suggests that miRNA’s are likely have a key role in human wound healing, possibly contributing to transitions between the successive phases of wound healing process; closure (reepithelialisation), maturation (squamous differentiation) and remodelling (scar remediation). Micro RNA’s can have both inductive and repressive activities.
This project seeks to indentify miRNA’s expressed by keratinocytes during the distinctive phases of the wound healing cascade. The student will harvest and cultivate keratinocytes from donated skin tissue discarded after surgical procedures and establish ex vivo skin equivalent tissue (HSE) models. RNA’s will be isolated from native donor human skin and from the HSE models under various conditions emulating tissue responses to wounding and healing. Initial experiments will focus on acute wound healing during the technology establishment phase; although it is aimed to apply this technology to investigate the delayed healing responses observed in patients with chronic / problem wounds. The student will be part of a multi–disciplinary team examining wider aspects of wound healing process and acquire advanced skills in cell biology and nucleic acid analysis. (Ref: Shaw & Martin (2009) EMBO Reports 10:881–886. doi:10.1038/embor.2009.102).
Methods and techniques developed
This project marries cell biology and nucleic acid technologies to discover if microRNA has a role during mammalian/human wound repair. Advanced RNA techniques including miRNA detection, microarray expression analysis RT–PCR and qRT–PCR, will be required and combined with classical histology and functional cell biology techniques such as organotypic skin culture (HSE), proliferation, apoptosis and migration assays, Western blotting, ELISA/EIA and immunohistochemistry.