Function of non-coding RNAs in liver cancer

我们的项目的目的是阐明长链非蛋白编码RNA（lncRNA）在最常见的肝癌形式，肝细胞癌（HCC）中的功能。我们希望了解肝癌发生的分子机制，这可能是未来的治疗方法，在我们以前的工作中，我们成功地发现了新的肝癌相关的lncRNA，通过编译他们的表达和调控在肝癌相比，正常肝组织的广泛景观。为了根据它们的功能重要性选择HCC相关的lncRNA，我们产生了靶向638个癌症相关lncRNA的3200个siRNA的文库。这个非常小但集中的文库应该是筛选调节癌症标志的lncRNA的通用工具。该项目的目的是筛选这种独特的siRNA文库中影响肝癌细胞增殖或凋亡的lncRNA。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进行深入的功能表征。此外，我们将进一步研究已知的lncRNA，我们已经确定了它们在HCC（例如HULC）中的作用。这些已知和新的命中分析包括功能丧失和获得模型的生成及其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-蛋白质-网络。基于观察到的表型、鉴定的相互作用伴侣和生物信息学的关联内疚预测，将形成和测试关于这些lncRNA的精确机制的假设。我们将在分子和细胞水平上阐明HULC和新的lncRNA在HCC中的作用，以了解lncRNA的基本机制，调节HCC中的肿瘤发生，并鉴定治疗靶基因以及调控网络。

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.