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.

废话介导的RNA衰减（NMD）是快速降解精选mRNA的机制。最近的研究已经发现NMD所需的UPF1基因在> 80％的腺泡胰腺癌（ASPC）是胰腺癌的一种特别激进的形式。我们有确定胰腺癌中的UPF1突变导致UPF1表达降低。其他突变最近报道的灭活NMD在胰腺导管腺癌中发现，我们已经报道说胰腺癌抑制NMD活性中常见的许多应力。 NMD抑制促进软琼脂，皮下外植体和原位胰腺移植中转化细胞的生长模型。我们的总体目标是更好地了解NMD抑制如何增加肿瘤的生长并探索如何我们可以利用NMD抑制胰腺癌的治疗增加。 RNA稳定性筛选，RNASEQ和代谢组学筛查已将Notch信号和糖酵解确定为 NMD调控途径。 Notch信号传导和糖酵解在胰腺癌中都起着重要作用一般和最近的胰腺癌分子分类研究表明，这两种途径是在ASPC中特别活跃，其中NMD通常在遗传上灭活。重要的是这些途径可以也是针对的。在AIM 1中，我们将确定NMD抑制Notch的机制和意义胰腺癌的激活。根据我们的初步数据，我们假设减少了NMD抑制作用表达稳定了Notch配体和受体，Notch信号的激活抑制E-钙粘蛋白表达在转移和化学抗性中起关键作用。但是我们还假设NMD 抑制胰腺癌将特别容易受到缺口抑制剂的影响。在AIM 2中，我们将确定如何 NMD抑制作用减少可调节代谢途径并利用它以获得治疗性增益。基于我们初步数据，我们假设NMD抑制稳定了编码的剪接成绩单线粒体呼吸系统的成员，这激活了糖酵解和五肽磷酸盐分流。这些途径的激活应使UPF1突变对临床更敏感可用的线粒体抑制剂和其他代谢抑制剂，如初步聚焦shRNA所示合成致死性筛选。对于这两个目标，我们都将使用各种体外细胞生物学，生化和分子技术。我们将使用独特的ASPC组织阵列来验证我们的体外发现以及一种新颖基因工程的小鼠，我们可以在胰腺中暂时下调UPF1的表达，而因此，可以忠实地对ASPC中的UPF1突变的后果进行建模。