Role of nonsense mediated RNA decay in pancreatic cancer

无义介导的RNA衰变在胰腺癌中的作用

基本信息

批准号：
9447641
负责人：
MARK Reid PHILIPS
金额：
$ 44.37万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9447641
关键词：
Affect Agar Alternative Splicing Biochemical Blood Vessels Carbohydrates Cell Respiration Cells Cellular Stress Cellular biology Classification Clinical Clustered Regularly Interspaced Short Palindromic Repeats Data Degradation Pathway Differentiation and Growth E-Cadherin Environment Enzymes Gene Expression Genes Genetic Transcription Genetically Engineered Mouse Glucose Glycolysis Goals Growth Human Hypoxia In Vitro KRAS2 gene Ligands Malignant Neoplasms Malignant neoplasm of pancreas Mediating Messenger RNA Metabolic Metabolic Pathway Metabolism MicroRNAs Mitochondria Modeling Molecular Multiprotein Complexes Mus Mutate Mutation Neoplasm Metastasis Nucleotide Biosynthesis Oxygen Pancreas Pancreatic Ductal Adenocarcinoma Pathway interactions Pentosephosphate Pathway Phenotype Play Process Proteins RNA Decay RNA Degradation RNA Helicase RNA Splicing RNA Stability Radiolabeled Receptor Activation Reporting Repression Resistance Respiration Role Shunt Device Signal Transduction Source Stress System Techniques Testing Therapeutic Tissue Microarray Transcript Transplantation Xenograft procedure base cell transformation epigenetic regulation genome wide screen in vitro Assay in vivo inhibitor/antagonist mRNA Expression mRNA Stability member metabolomics migration mitochondrial messenger RNA notch protein novel nutrient deprivation pancreatic cancer cells pancreatic neoplasm small hairpin RNA subcutaneous targeted treatment transcriptome transcriptome sequencing transplant model tumor tumor growth

项目摘要

Nonsense mediated RNA decay (NMD) is a mechanism to rapidly degrade select mRNAs. Recent studies have found that the UPF1 gene, required for NMD, is strikingly mutated and inactivated in >80% of adenosquamous pancreatic cancer (ASPC), a particularly aggressive form of pancreatic cancer. We have determined that UPF1 mutations in pancreatic cancer result in decreased UPF1 expression. Other mutations recently reported to inactivate NMD are found in pancreatic ductal adenocarcinoma, and we have reported that many of the stresses commonly found in pancreatic cancer repress NMD activity. NMD inhibition promotes the growth of transformed cells in soft agar, subcutaneous explants, and in an orthotopic pancreatic transplant model. Our overall goal is to better understand how NMD inhibition augments tumor growth and explore how we can exploit NMD inhibition for therapeutic gain in pancreatic cancer. RNA stability screens, RNAseq, and metabolomics screens have identified Notch signaling and Glycolysis as NMD regulated pathways. Both Notch signaling and Glycolysis play an important role in pancreatic cancer in general, and recent pancreatic cancer molecular classification studies indicate that these two pathways are particularly active in ASPC, where NMD is typically genetically inactivated. Importantly these pathways can also be targeted. In Aim 1 we will identify the mechanism and significance of NMD inhibition on Notch activation in pancreatic cancer. Based on our preliminary data we hypothesize that reduced NMD inhibition expression stabilizes Notch ligands and receptors, and the activation of Notch signaling represses e-cadherin expression to play a key role in metastases and chemo-resistance. However we also hypothesize that NMD inhibited pancreatic cancers will be particularly susceptible to Notch inhibitors. In Aim 2 we will determine how reduced NMD inhibition regulates metabolic pathways and exploit this for therapeutic gain. Based on our preliminary data, we hypothesize that NMD inhibition stabilizes alternatively spliced transcripts encoding members of the mitochondrial respiration system, and this activates glycolysis and the pentose phosphate shunt. The activation of these pathways should render tumors with UPF1 mutations more sensitive to clinically available mitochondrial inhibitors and other metabolic inhibitors, as indicated by preliminary focused shRNA synthetic lethality screens. For both Aims we will use a variety of in vitro cell biology, biochemical, and molecular techniques. We will validate our in vitro findings with unique ASPC tissue arrays, as well as a novel genetically engineered mouse in which we can temporally down-regulate UPF1 expression in pancreas, and can thus faithfully model the consequences of UPF1 mutations found in ASPC.

无义介导的 RNA 降解 (NMD) 是一种快速降解选定 mRNA 的机制。最近的研究发现 NMD 所需的 UPF1 基因在超过 80% 的人中发生显着突变和失活腺鳞状胰腺癌（ASPC），一种特别具有侵袭性的胰腺癌。我们有确定胰腺癌中 UPF1 突变导致 UPF1 表达减少。其他突变最近有报道称，在胰腺导管腺癌中发现了 NMD 失活，我们报道称胰腺癌中常见的许多应激都会抑制 NMD 活性。 NMD 抑制促进转化细胞在软琼脂、皮下外植体和原位胰腺移植中的生长模型。我们的总体目标是更好地了解 NMD 抑制如何促进肿瘤生长并探索如何我们可以利用 NMD 抑制来获得胰腺癌的治疗效果。 RNA 稳定性筛选、RNAseq 和代谢组学筛选已将 Notch 信号传导和糖酵解确定为 NMD 调控途径。 Notch 信号传导和糖酵解在胰腺癌中发挥着重要作用一般而言，最近的胰腺癌分子分类研究表明这两条途径是在 ASPC 中特别活跃，其中 NMD 通常是基因失活的。重要的是，这些途径可以也有针对性。在目标 1 中，我们将确定 NMD 抑制 Notch 的机制和意义胰腺癌中的激活。根据我们的初步数据，我们假设减少了 NMD 抑制表达稳定 Notch 配体和受体，Notch 信号传导的激活抑制 e-钙粘蛋白表达在转移和化疗耐药性中发挥关键作用。然而我们也假设 NMD 受到抑制的胰腺癌对 Notch 抑制剂特别敏感。在目标 2 中，我们将确定如何减少 NMD 抑制可调节代谢途径并利用其获得治疗效果。基于我们的根据初步数据，我们假设 NMD 抑制可稳定编码的可变剪接转录本线粒体呼吸系统的成员，这会激活糖酵解和磷酸戊糖分流。这些途径的激活应该使具有 UPF1 突变的肿瘤对临床更加敏感。可用的线粒体抑制剂和其他代谢抑制剂，如初步聚焦的 shRNA 所示合成致死率筛选。对于这两个目标，我们将使用各种体外细胞生物学、生物化学和分子技术。我们将使用独特的 ASPC 组织阵列以及一种新颖的方法来验证我们的体外研究结果基因工程小鼠，我们可以暂时下调胰腺中 UPF1 的表达，以及因此可以忠实地模拟 ASPC 中发现的 UPF1 突变的后果。