Screening Assays for Small Molecules Targeting Oncogenic eIF4E Expression

针对致癌 eIF4E 表达的小分子筛选试验

基本信息

批准号：
8435372
负责人：
Chunhong Yan
金额：
$ 4.04万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2013-09-13
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8435372
关键词：
Actins Address Animals Antineoplastic Agents Antisense Oligonucleotides Automobile Driving BCL2 gene Biological Assay Bioluminescence CMV promoter Cancer Cell Growth Cell Line Cell Survival Cells Chemicals Chromatin Clinical Trials DNA Dependovirus Development Disadvantaged Distal Down-Regulation Drug Targeting Emerging Technologies Environment Eukaryotic Initiation Factor-4E Eukaryotic Initiation Factors Firefly Luciferases Galactosidase Gene Expression Genes Genetic Transcription Genomics Guidelines Health Human Hyperactive behavior Internal Ribosome Entry Site Lead Location Luc Gene Luciferases Malignant Neoplasms Measurement Measures Mediating Messenger RNA Methods NIH Program Announcements Oncogenic PI3K/AKT Pathway interactions Pharmaceutical Preparations Preclinical Drug Evaluation Promoter Regions Proteins RNA Cap-Binding Proteins Regulatory Element Renilla Luciferases Reporter Reporter Genes Research Robotics Technology Testing Therapeutic Therapeutic Agents Therapeutic Intervention Toxic effect Transcript Transgenic Organisms Translations Untranslated Regions Vascular Endothelial Growth Factors assay development base c-myc Genes cancer cell cancer therapy cell growth cellular engineering cost drug development drug discovery high throughput screening homologous recombination human FRAP1 protein in vivo inhibitor/antagonist innovation luminescence neglect novel overexpression promoter recombinase research clinical testing response screening small molecule small molecule libraries success tumor growth

项目摘要

DESCRIPTION (provided by applicant): The eukaryotic translation initiation factor 4E (eIF4E) is frequently overexpressed in human cancers and contributes to cancer development and progression by selectively driving translation of genes that are essential for cancer cell growth and survival (e.g., c-myc, VEGF, BCL-2). In the past decades accumulating evidence has demonstrated that down-regulation of eIF4E expression in cancer cells can be an effective strategy for therapeutic intervention of cancer. Indeed, eIF4E antisense oligonucleotides have been shown to inhibit tumor growth without toxicity to host animals, and have entered clinical trials. However, because of the lack of a reliable assay that can be used in high-throughput drug screening, no small molecule that can inhibit eIF4E expression in cancer cells is currently available for clinical testing. This application is in response to the NIH Program Announcement PA-10-213, "Development of Assays for High-Throughput Screening for Use in Probe and Pre-therapeutic Discovery", and will address the imperative need of a high-throughput screening (HTS) assay for discovery of drug leads that target eIF4E expression in cancer cells. Specifically, our objective is to utilize emerging cell-engineering technologies to develop an innovative assay in which a bioluminescent reporter integrated in the native eIF4E gene locus is expressed under control of the endogenous eIF4E promoter. Such an assay is expected to overcome limitations of current reporter assays that are mostly based on cloned, transgenic promoters, which often lack essential distal/intronic cis-regulatory elements while residing in foreign chromatin environments. Therefore, the innovative assay would faithfully reproduce responses of the endogenous eIF4E gene to chemical treatments and thus be more reliable in high-throughput search for small molecules inhibitory for eIF4E expression. Accordingly, the specific aims of this project are: (1) to develop cell lines harboring a reporter gene in the native eIF4E gene locus for high-throughput drug screening, (2) to develop secondary assays for identification of false positives and prioritization of screening hits for further drug development. Completion of the proposed research will result in an innovative HTS assay useful for developing therapeutic agents that can cure cancer through inhibiting eIF4E expression. Therefore, the proposed studies will impact health of millions of people afflicted with cancer. This application will also provide a proof of concept for an innovative drug-discovery strategy that can be applied to search for agents targeting any given gene aberrantly expressed in cancer thereby significantly advancing targeted cancer therapy.

描述（由申请人提供）：真核翻译起始因子4E（EIF4E）经常在人类癌症中过表达，并通过选择性地推动对癌细胞生长和生存至关重要的基因的转化来促进癌症的发展和进展（例如C-Myc，VEGF，BCL-2）。在过去的几十年中，积累的证据表明，癌细胞中EIF4E表达的下调可能是癌症治疗干预的有效策略。实际上，EIF4E反义寡核苷酸已被证明可抑制肿瘤的生长而没有毒性的动物，并且已经进行了临床试验。但是，由于缺乏可用于高通量药物筛查的可靠测定法，目前可用于临床测试，没有可抑制癌细胞中EIF4E表达的小分子。该应用是对NIH计划公告PA-10-213的响应，“开发用于用于探针和治疗前发现的高通量筛选的测定”，并将解决对高通量筛查（HTS）测定的必要需求，以发现靶向EIF4E在癌细胞中靶向EIF4E表达的药物潜在客户。具体而言，我们的目标是利用新兴的细胞工程技术来开发一种创新的测定法，其中在本地EIF4E基因基因座中积分的生物发光记者在内源性EIF4E启动子的控制下表达。这种测定有望克服当前报告基因测定的局限性，这些测定主要基于克隆的转基因启动子，这些启动子通常缺乏居住在异物染色质环境的同时，通常缺乏必不可少的远端/内含子顺式调节元件。因此，创新的测定法将忠实地再现内源性EIF4E基因对化学处理的反应，因此在高通量搜索抑制EIF4E表达的小分子时更可靠。因此，该项目的具体目的是：（1）在天然EIF4E基因基因座中开发具有记者基因的细胞系，以进行高通量药物筛查，（2）开发次要测定，以鉴定错误的阳性和筛查筛查的优先次序以进一步药物开发。拟议研究的完成将导致创新的HTS测定法，可用于开发可以通过抑制EIF4E表达来治愈癌症的治疗剂。因此，拟议的研究将影响数百万患有癌症的人的健康。该应用还将为创新的药物发现策略提供概念证明，该策略可用于搜索针对癌症中任何给定基因的药物，从而显着提高靶向癌症治疗。