Function of non-coding RNAs in liver cancer

我们项目的目的是阐明长链非蛋白编码rna （lncRNA）在最常见的肝癌——肝细胞癌（HCC）中的功能。我们希望了解肝癌发生的分子机制，为未来的治疗方法提供可能。在我们之前的工作中，我们成功地发现了新的HCC相关的lncrna，通过编制其在HCC中的表达和调控的广泛景观与正常肝组织相比。为了根据功能重要性选择hcc相关的lncrna，我们生成了一个包含3200个sirna的文库，目标是638个癌症相关的lncrna。这个相对较小但重点突出的文库应该是筛选调节癌症特征的lncrna的通用工具。本项目旨在筛选这一独特的siRNA文库，寻找影响肝癌细胞增殖或凋亡的lncRNAs。然后将这些功能数据与这组癌症相关lncrna的表达和调控数据相结合，以选择最有希望的命中点进行深入的功能表征。此外，我们将进一步研究已知的lncrna，这些lncrna已经在HCC中发挥了作用（例如HULC）。对这些已知的和新颖的hit的分析包括功能损失和功能获得模型的生成以及它们在细胞和分子水平上的表征。创新的基因组编辑技术将用于在人类HCC细胞中创建lncrna的定量敲除。基于大多数lncrna将作为核糖核蛋白复合物的假设，我们将使用RNA亲和纯化来揭示lncrna -蛋白网络。基于观察到的表型、确定的相互作用伙伴和生物信息学上的关联内疚预测，将形成和检验这些lncrna的精确机制的假设。总之，我们将在分子和细胞水平上阐明HULC和新型lncrna在HCC中的重要作用，以了解lncrna调节的HCC肿瘤发生的基本机制，并确定治疗靶基因和调控网络。

The aim of our project is to elucidate the function of long non-protein-coding RNAs (lncRNA) in the most frequent form of liver cancer, hepatocellular carcinoma (HCC). We want to understand the molecular mechanisms underlying liver carcinogenesis which might be exploitable for future therapeutic approaches.In our previous work, we successfully discovered novel HCC-associated lncRNAs by compiling a broad landscape of their expression and regulation in HCC compared to normal liver tissue. To select HCC-associated lncRNAs based on their functional importance, we generated a library of 3200 siRNAs targeting 638 cancer-associated lncRNAs. This comparably small, but focused library should be a versatile tool to screen for lncRNAs modulating the hallmarks of cancer. The proposed project aims to screen this unique siRNA library for lncRNAs affecting the proliferation or apoptosis of liver cancer cells. These functional data are then combined with the expression and regulation data for this set of cancer-associated lncRNAs to select the most promising hits for an in-depth functional characterization. In addition, we will further pursue our studies of known lncRNAs for which we have already established a role in HCC (e.g. HULC). The analysis of these known and novel hits includes the generation of loss- and gain-of-function models and their characterization at the cellular and molecular level. Innovative genome editing techniques will be used to create quantitative knockouts of lncRNAs in human HCC cells. Based on the hypothesis that most lncRNAs will act as ribonucleoprotein complexes, we will uncover the lncRNA-protein-network using RNA affinity purifications. Hypotheses on the precise mechanisms of these lncRNAs will be formed and tested based on the observed phenotypes, the identified interaction partners and bioinformatical guilt-by-association predictions.In summary, we will unravel the role of HULC and novel lncRNAs functionally important in HCC at the molecular and cellular level to understand fundamental mechanisms of lncRNA-modulated tumorigenesis in HCC and to identify therapeutic target genes as well as regulatory networks.