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

长非编码RNAs(Long Non-Coding RNAs，LncRNAs)在基因组中具有重要的调节作用，许多LncRNAs在人类疾病过程中被发现是失控的。LncRNAs是一类非编码基因，不编码蛋白质，长度超过200bp。据推测，lncRNAs将在神经脊分化、黑素细胞发育以及人类黑色素瘤的发生过程中发挥重要的调节作用。本研究的目的是鉴定和鉴定MITF-Sox10调控的位置保守的lncRNAs在人类和斑马鱼基因组中在神经脊分化、黑素细胞发育和人类黑色素瘤过程中的功能。黑色素瘤是一种侵袭性皮肤癌，起源于一种称为黑素细胞的皮肤细胞的恶性转化，一旦转移与预后不良有关。将表型转换为去分化的神经脊干细胞样状态被认为有助于黑色素瘤的治疗耐药。在发育过程中，神经脊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方案，以提供新的治疗目标和改善预后。研究将集中于在人类(智人)和斑马鱼(Danio Rerio)基因组中识别位置保守的MITF-Sox10调控的lncRNAs(目标1)。MITF和Sox10是两种转录因子，它们调节涉及神经脊分化、黑素细胞发育和人类黑色素瘤的基因表达程序。这是必须的第一步，它将为黑色素瘤的发展和致癌事件中控制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的lncRNA。为了确定候选的lncRNAs在神经脊分化和黑素细胞发育过程中的功能，将在斑马鱼中减弱lncRNAs的表达作为体内模型(目标2)。使用相关的体内模型将允许识别在不同发育阶段对神经脊分化和黑素细胞发育至关重要的lncRNAs。通过这些实验，可以对不同LncRNA的分子功能进行评估和表征。最后，为了确定候选的lncRNA是否在临床上与人类健康相关，我们在体外减弱了人黑色素瘤细胞系中lncRNA的表达(目标3)。通过这一目标，可以评估黑色素瘤背景下lncRNAs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lncRNAs的功能，以及它们影响神经脊分化、黑素细胞发育和黑色素瘤的基因表达程序的分子机制。这项拟议的工作体现了BBSRC的优先领域之一：“生物科学促进对健康的综合理解”。

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'.