Investigating positionally conserved long non-coding RNA functions during melanocyte development in zebrafish and human melanoma.

Long non-coding RNAs (lncRNAs) have important regulatory roles within the genome with many lncRNAs found to be dysregulated during human disease. LncRNAs are a class of non-coding genes that do not code for proteins and are longer than 200bp in length. It is hypothesised that lncRNAs will play a vital regulatory role in neural crest differentiation, melanocyte development and consequently, human melanoma. The aim of this research project is to identify and characterise the function of positionally conserved lncRNAs regulated by MITF-SOX10 between human and zebrafish genomes during neural crest differentiation, melanocyte development and human melanoma.

Melanoma is an aggressive skin cancer which derives from the malignant transformation of a type of skin cell called melanocytes and is associated with a poor prognosis once metastasised. Switching phenotypes to a de-differentiated neural crest stem cell-like state is hypothesised to contribute to therapeutic resistance observed in melanoma. During development, neural crest cells give rise to several types of cells including melanocytes. Highlighting the need for further understanding of the molecular mechanisms which govern the progression of melanoma as well as their associated developmental programmes to provide new therapeutic targets and improved prognosis.

Research will focus on identifying positionally conserved MITF-SOX10 regulated lncRNAs in human (Homo sapiens) and zebrafish (Danio rerio) genomes (Objective 1). MITF and SOX10 are two transcription factors that regulate gene expression programmes involved in neural crest differentiation, development of melanocytes and human melanoma. This is an imperative initial step which will provide novel insights into the molecular mechanisms governing gene expression programmes during development and oncogenic events in melanoma, providing candidate lncRNAs to prioritise for further research.

To ascertain the function of candidate lncRNAs during neural crest differentiation and melanocyte development, expression of lncRNAs will be attenuated in zebrafish as an in-vivo model (Objective 2). The use of a relevant in-vivo model will allow for the identification of lncRNAs important for neural crest differentiation and melanocyte development at different developmental stages. Through these experiments, molecular function of different lncRNAs can be assessed and characterised.

Finally, to ascertain whether candidate lncRNAs identified are clinically relevant in human health, lncRNA expression be attenuated in-vitro in human melanoma cell lines (Objective 3). Through this objective, the impact of lncRNAs in human health in the context of melanoma can be assessed. Providing new therapeutic targets and biomarkers that can be used to improve melanoma therapeutics. The outcomes of these objectives will better our understanding of the function of lncRNAs and the molecular mechanisms that they govern to influence gene expression programmes for neural crest differentiation, melanocyte development and melanoma. This proposed work embodies one of BBSRC's priority areas: 'Bioscience for an Integrated Understanding of Health'.