Exploration of long non-coding RNAs as synthetic essential targets in Pten-deficient cancers

探索长非编码 RNA 作为 Pten 缺陷癌症的合成必需靶标

基本信息

批准号：
10321527
负责人：
Jasper R Chen
金额：
$ 4.43万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-18 至 2025-09-17
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10321527
关键词：
ANXA5 gene Acetylation Address Affect Alleles Aneuploidy Apoptosis Apoptotic Binding Sites Biological Assay Bromodeoxyuridine CASP3 gene Categories Cell Cycle Cell Death Cell Line Cell Proliferation Cell Survival Cells ChIP-seq Characteristics Chromatin Clinical Code Colcemid Complex DNA Damage Development Doxycycline Ensure Essential Genes Feedback Gene Expression Gene Silencing Gene set enrichment analysis Genes Genetic Transcription Genetically Engineered Mouse Genome Genome Stability Genomic Instability Glioblastoma Glioma Gliomagenesis Goals Growth and Development function Histone Acetylation Histone H3 Histone H4 Homologous Gene Impairment Knock-out Knowledge Lead Lysine MYC Family Protein Malignant Neoplasms Mass Spectrum Analysis Mediating Modeling Mouse Cell Line Mus Mutagens Mutation Neuraxis PI3K/AKT PTEN gene Pathway interactions Patient-Focused Outcomes Pattern Pharmaceutical Preparations Physicians Play Proteins RNA purification Radiation therapy Regulation Research Resources Role SAGA Sampling Science Scientist Signal Transduction Site Suggestion T47D Tamoxifen Testis The Cancer Genome Atlas Tissues Training Transactivation Treatment outcome Tumor Burden Tumor Suppressor Genes Untranslated RNA Western Blotting Work cancer cell cancer genome cancer genomics cancer survival cancer therapy cancer type career cell type chromatin isolation by RNA purification sequencing chromatin remodeling frontier genotoxicity histone acetyltransferase improved in vivo loss of function mouse model nestin protein novel novel therapeutics outreach overexpression promoter recruit targeted treatment tumor tumorigenesis

项目摘要

Project Summary/Abstract PTEN is one of the most frequently inactivated tumor suppressor genes across all cancer types. The loss of PTEN activates PI3K/AKT, which inhibits GSK3β, thereby stabilizing Myc and contributing to oncogenesis. Myc recruits histone acetyltransferases to increase chromatin accessibility of target genes involved in both cell proliferation and apoptosis. Among these histone acetyltransferases, the Spt-Ada-Gcn5 acetyltransferase (SAGA) complex preferentially acetylates histone H3 lysine 9 and histone H4 lysine 16 to activate gene expression. A pan-cancer analysis of mutually exclusive gene inactivation patterns identified a previously uncharacterized long non-coding RNA (lncRNA) as synthetic essential in the context of PTEN deficient cancer. Preliminary studies suggest that this lncRNA inhibits SAGA-mediated histone acetylation, thereby inhibiting Myc transactivation of target genes. PTEN and lncRNA double-knockout SF-763 glioma cells showed Myc pathway enrichment, impaired cell viability, and pronounced aneuploidy, which was not observed in wild-type or single knockout cell lines. We hypothesize that inhibition of SAGA-mediated acetylation by this lncRNA inhibits Myc transactivation of pro-apoptotic target genes and Myc-driven endoreduplication, thereby promoting cancer survival. This proposal will investigate the potential of targeting the poorly studied non-coding genome for cancer treatment, advance our knowledge of the role of histone acetylation on cancer genomic stability (widely targeted using genotoxic drugs and radiotherapy), and describe a novel mechanism for the regulation of Myc's dual functions in proliferation and apoptosis. We will verify lncRNA expression in cell lines from various cancer types and in clinical samples to validate the pan-cancer relevance and translational potential of this study, respectively. Annexin V and caspase 3/7 assays will be used to assess the hypothesis of Myc-driven apoptosis. Endoreduplication will also be probed using BrdU incorporation into colcemid-arrested cells. Chromatin isolation by RNA purification and chromatin immunoprecipitation sequencing will be used to demonstrate how the lncRNA inhibits histone acetylation by SAGA. To assess the role of the lncRNA in vivo, we will functionally validate its putative mouse homolog and generate a genetically engineered mouse model to characterize its effects on tumor development. The lncRNA knockout allele will be bred into a Qki;Pten;Trp53 glioblastoma mouse model to assess the effects of lncRNA suppression in the context of Pten deletion. The training plan will address gaps in the applicant's research and clinical abilities, ensuring that he can successfully complete the proposed work and preparing him for the next stage of his career. The training will be completed in Dr. Ronald DePinho's lab at MD Anderson, where the applicant will have access to the resources, facilities, and, most importantly, colleagues that will nurture his continuing development and growth into a physician-scientist.

项目摘要/摘要 PTEN是所有癌症类型中最常见的肿瘤抑制基因之一。损失 PTEN激活抑制GSK3β的PI3K/AKT，从而稳定MYC并导致肿瘤发生。妈妈招募组蛋白乙酰转移酶以增加涉及两个细胞的靶基因的染色质可及性增殖和凋亡。在这些组蛋白乙酰转移酶中，SPT-ADA-GCN5乙酰转移酶（传奇）复合物优先乙酰化组蛋白H3赖氨酸9和组蛋白H4赖氨酸16激活基因表达。互斥基因失活模式的泛伴奏分析确定了先前的在PTEN缺乏癌症的背景下，未表征的长期非编码RNA（LNCRNA）是合成的。初步研究表明，该LNCRNA抑制了传奇介导的组蛋白乙酰化，从而抑制了MYC 靶基因的反式激活。 PTEN和LNCRNA双敲除SF-763胶质瘤细胞显示了MYC途径富集，细胞活力受损并发音为非整倍性，在野生型或单个中未观察到它敲除单元线。我们假设该lncRNA抑制传奇介导的乙酰化抑制MYC 促凋亡靶基因和MYC驱动的内置的反式激活，从而促进癌症生存。该提案将研究针对癌症研究不足的非编码基因组的潜力治疗，提高我们对组蛋白乙酰化对癌症基因组稳定性的作用的了解（广泛针对性）使用遗传毒性药物和放射疗法），并描述一种调节MYC双重的新机制在增殖和凋亡中起作用。我们将验证来自各种癌症类型的细胞系中的lncRNA表达并在临床样本中分别验证泛伴侣的相关性和翻译本研究的潜力。膜联蛋白V和caspase 3/7分析将用于评估MYC驱动的凋亡的假设。还将使用将BRDU纳入结构性捕获的细胞中的brdu进行探测。染色质分离通过RNA纯化和染色质免疫沉淀测序将用于证明LNCRNA如何通过传奇抑制组蛋白乙酰化。为了评估lncRNA在体内的作用，我们将在功能上验证其假定的小鼠同源物并生成一般设计的鼠标模型，以表征其对肿瘤的影响发展。 lncRNA敲除等位基因将繁殖到QKI； pten； trp53胶质母细胞瘤小鼠模型到在PTEN缺失的背景下评估LNCRNA抑制的影响。培训计划将解决差距申请人的研究和临床能力，确保他可以成功完成拟议的工作和为他的职业生涯的下一个阶段做准备。该培训将在MD的Ronald Depinho博士的实验室完成安德森（Anderson），申请人将可以访问资源，设施，最重要的是同事这将为他的持续发展和成长提供给身体科学家的持续发展。