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

长链非编码rna （lncRNAs）在基因组中具有重要的调控作用，许多lncRNAs在人类疾病中被发现失调。LncRNAs是一类不编码蛋白质的非编码基因，长度超过200bp。据推测，lncrna将在神经嵴分化、黑素细胞发育以及人类黑色素瘤中发挥重要的调节作用。本研究项目的目的是鉴定和表征MITF-SOX10在人类和斑马鱼基因组中在神经嵴分化、黑素细胞发育和人类黑色素瘤中调控的位置保守lncRNAs的功能。黑色素瘤是一种侵袭性皮肤癌，源于一种叫做黑素细胞的皮肤细胞的恶性转化，一旦转移，预后就很差。将表型转换为去分化的神经嵴干细胞样状态被假设有助于在黑色素瘤中观察到的治疗抗性。在发育过程中，神经嵴细胞产生几种类型的细胞，包括黑素细胞。强调需要进一步了解控制黑色素瘤进展的分子机制及其相关的发展计划，以提供新的治疗靶点和改善预后。研究将集中于鉴定人类（智人）和斑马鱼（达尼欧河）基因组中位置保守的MITF-SOX10调控lncrna（目的1）。MITF和SOX10是调节神经嵴分化、黑色素细胞发育和人类黑色素瘤的基因表达程序的两个转录因子。这是至关重要的第一步，将为黑素瘤发展和致癌事件过程中调控基因表达程序的分子机制提供新的见解，为进一步研究提供候选lncrna的优先级。为了确定候选lncRNAs在神经嵴分化和黑素细胞发育过程中的功能，将在斑马鱼体内模型中降低lncRNAs的表达（目的2）。使用相关的体内模型将允许鉴定在不同发育阶段对神经嵴分化和黑素细胞发育重要的lncrna。通过这些实验，可以评估和表征不同lncrna的分子功能。最后，为了确定鉴定出的候选lncRNA是否与人类健康有临床相关性，我们在体外降低了lncRNA在人类黑色素瘤细胞系中的表达（目的3）。通过这一目标，可以评估lncrna在黑色素瘤背景下对人类健康的影响。提供新的治疗靶点和生物标志物，可用于改善黑色素瘤的治疗。这些目标的结果将使我们更好地理解lncrna的功能及其影响神经嵴分化、黑素细胞发育和黑色素瘤的基因表达程序的分子机制。这项提议的工作体现了BBSRC的优先领域之一：“生物科学对健康的综合理解”。

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'.