Understanding the role of ribosome-associated long non-coding RNA in cancers of the neural crest

了解核糖体相关长非编码 RNA 在神经嵴癌中的作用

• 批准号: 2879751
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2879751
• 关键词: Understanding; role; ribosome; associated; long

Whilst 85% of our genome is transcribed, only 4% is protein-coding. In fact, many non-coding RNAs have key roles in gene regulation and are termed long non-coding RNAs (lncRNAs). These exhibit far more tissue and developmental stage specific expression than mRNAs and are specifically enriched in the nervous system. Diseases of the nervous system are particularly associated with lncRNAs mis-regulation conditions e.g. Alzheimer's disease. The majority of characterised lncRNAs are nuclear but our recent Ribo-Ribo-Seq in human neuroblastoma cell line has revealed that many are cytoplasmic. We have discovered 278 cytoplasmic lncRNAs regulated during neuronal differentiation. 43% of these lncRNAs are associated with ribosomes but they themselves are not translated. We hypothesise that these ribosome-associated lncRNAs regulate mRNA translation in cancer. This project seeks to understand the molecular mechanisms of this translational regulation by lncRNAs during neuronal differentiation and the role these lncRNA play in cancers of the neural crest (e.g. neuroblastoma).Objectives1-Assess the importance of ribosome-associated lncRNAs to neuronal differentiation in neuroblastoma cell lines. 2-Determine the effect on mRNA translation by ribosome-associated lncRNAs. 3-Dissect molecular mechanisms of translational regulation by ribosome-associated lncRNAs. 4-Probe dysregulation of ribosome-associated lncRNAs in neuroblastoma. NoveltyThe project will take a range of cutting-edge and interdisciplinary approaches to understand the function of neuronal lncRNAs and their contribution to cancers of the nervous system including neuroblastoma. By combining Next Generation Sequencing, structural biology, patient derived primary cells and cell biology, we will take a novel holistic view of neuronal lncRNA-protein biology.TimelinessOnly a small number of lncRNAs have been functionally characterised and the main focus has been on nuclear lncRNAs. Our recent work has identified this novel population of cytoplasmic lncRNAs which associate with ribosomes. The importance of these lncRNA-ribosome interactions has yet to be determined. RNA pull-down approaches have also recently been developed to isolate lncRNA-protein complexes from cells and tissues. Experimental Approaches-CRISPR/siRNA of lncRNA in neuroblastoma cell line and patient-derived neuroblastoma primary cultures-Translation profiling by polysome gradients / Ribo-Seq-Biochemical purification of lncRNA-ribosome complexes followed by cryo-EM-Single molecule fluorescence microscopy of lncRNA in patient derived and culture neural crest cells-Cell proliferation/ migration assays in neuroblastoma cell lines and primary cells-Analysis of publicly available cancer data on lncRNA expressed and dysregulation.

尽管我们的基因组的85％被转录，但只有4％是蛋白质编码。实际上，许多非编码RNA在基因调节中具有关键作用，称为长的非编码RNA（LNCRNA）。与mRNA相比，这些表现出更多的组织和发育阶段的特异性表达，并且特别富含神经系统。神经系统的疾病特别与LNCRNA的错误调节状况有关，例如阿尔茨海默氏病。大多数特征性的LNCRNA都是核的，但是我们最近在人类神经母细胞瘤细胞系中的核糖 - 核糖表明，许多人都表明许多是细胞质的。我们发现了在神经元分化过程中调节的278个细胞质LNCRNA。这些LNCRNA中有43％与核糖体相关，但它们本身并未翻译。我们假设这些核糖体相关的LNCRNA调节癌症中的mRNA翻译。该项目旨在了解LNCRNA在神经元分化过程中的这种翻译调节的分子机制，以及这些lncRNA在神经粉的癌症中的作用（例如，神经母细胞瘤）.Obigntives1-评估与核糖体相关的LNCRNAS对神经负分分化神经元分化细胞的重要性。 2确定对核糖体相关的LNCRNA对mRNA翻译的影响。核糖体相关LNCRNA的转化调节的3截止分子机制。神经母细胞瘤中与核糖体相关的LNCRNA的4探针失调。 Novelty The项目将采用一系列尖端和跨学科的方法，以了解神经元LNCRNA的功能及其对包括神经母细胞瘤在内的神经系统的癌症的贡献。通过结合下一代测序，结构生物学，患者衍生的原代细胞和细胞生物学，我们将对神经元LNCRNA-蛋白质生物学的整体视野进行新颖的整体视野。从功能上讲，少数LNCRNA在功能上表征了，主要的重点是核LNCRNA。我们最近的工作确定了与核糖体相关的新型细胞质lncRNA人群。这些LNCRNA-核糖体相互作用的重要性尚未确定。最近还开发了RNA下拉方法，以分离细胞和组织的lncRNA-蛋白质复合物。 Experimental Approaches-CRISPR/siRNA of lncRNA in neuroblastoma cell line and patient-derived neuroblastoma primary cultures-Translation profiling by polysome gradients / Ribo-Seq-Biochemical purification of lncRNA-ribosome complexes followed by cryo-EM-Single molecule fluorescence microscopy of lncRNA in patient derived and culture neural crest cells-Cell proliferation/ migration在神经母细胞瘤细胞系中的测定和原代细胞分析表达和失调的LNCRNA的癌症数据。