Pharmacological targeting of the Unfolded Protein Response as an antitumor strate

未折叠蛋白反应的药理学靶向作为抗肿瘤策略

基本信息

批准号：
7644769
负责人：
ALBERT KOONG
金额：
$ 34.55万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-17 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7644769
关键词：
5&apos Untranslated Regions Animals Anoxia Attenuated Biological Assay Biological Models Bortezomib Cell Culture Techniques Cell Survival Cells Cellular Assay Cellular Stress Clinical Collaborations Cytotoxic agent Data Defect Development Effectiveness Embryo Endoplasmic Reticulum Environment Exhibits Genes Glucose Goals Growth Growth Factor Health Human Hypoxia In Vitro Institution Kinetics Label Luciferases Malignant Neoplasms Measures Metabolic stress Modality Modeling Molecular Molecular Chaperones Multiple Myeloma Mus Mutate Oncogenic Oxygen Pathway interactions Phenotype Phosphotransferases Physiological Physiology Play Preclinical Drug Evaluation Process Property Proteasome Inhibitor Proteins Publishing Reporter Resistance Role Screening procedure Solid Neoplasm Stress System Testing Therapeutic Time Toxic effect Transgenic Mice Transgenic Organisms Translations Transplantation Treatment Efficacy Tumor Burden Up-Regulation Upper arm Work Xenograft procedure Zebrafish angiogenesis antitumor agent base cell transformation chemotherapeutic agent cytotoxic design efficacy testing endonuclease endoplasmic reticulum stress improved in vivo inhibitor/antagonist killings neoplastic cell novel novel strategies pre-clinical programs public health relevance research study response small molecule small molecule libraries tumor tumor growth tumor progression tumor xenograft tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Hypoxia/anoxia is a well-characterized component of the microenvironment of most solid tumor. Considerable experimental and clinical evidence supports the notion that hypoxia fundamentally alters the physiology of the tumor towards a more aggressive phenotype. The Unfolded Protein Response (UPR) is a cellular stress adaptation pathway which promotes cell survival in the presence of Endoplasmic Reticulum (ER) stress, including physiological stress in the tumor microenvironment. The PERK/eIF2a/ATF4 pathway reduces the global rates of protein translation thereby alleviating metabolic stress under hypoxia while at the same time induces the translational upregulation of important chaperones and pro-survival genes. Another UPR component is the activation of the endonuclease/kinase IRE1 and its immediate effector XBP1, which activate a transcriptional program aimed at increasing the folding capacity of the ER. Recent in vitro and in vivo studies from the labs of the two co-PIs, have shown that transformed cells with ablated UPR responses exhibit reduced tolerance to hypoxia in vitro and form tumors that are slower growing compared to tumors grown from cells with an intact UPR, indicating that UPR activation contributes to tumorigenesis. In preliminary studies, we have developed cell-based and animal-based assays for UPR activation and its inhibition by small molecules. The overall goal of this proposal is to validate UPR activation as an important anti- tumor target and to use novel in vitro and in vivo assays to identify potent inhibitors of this response as novel chemotherapeutic agents. In Aim 1, we will employ in vitro reporter assays of PERK activity to identify specific inhibitors of the PERK/eIF21/ATF4 pathway and test the effect of combined administration of these inhibitors with inhibitors of the IRE-1 pathway (Irestatins) on tumor cell survival under normoxia and hypoxia. In Aim 2, we will investigate the potential synergy between inhibitors of the PERK and IRE1 pathways with the proteasome inhibitor Bortezomib in killing hypoxic tumor cells in vitro and in vivo. Studies under Aim 3, will evaluate the use of Zebrafish as a model system to analyze the ability of inhibitors of the PERK and IRE1 pathways to inhibit xenotransplanted human tumors and to inhibit angiogenesis without causing significant developmental abnormalities. Finally, in Aim 4 we will test the efficacy and potential toxicity of identified compounds in mouse tumor models. We expect that these efforts will culminate in the development of specific and potent inhibitors of the UPR which alone, or in combination with existing antitumor agents and modalities will be effective in reducing tumor burden in preclinical and clinical malignancies. PUBLIC HEALTH RELEVANCE: A hallmark of solid tumors is the requirement to adapt to, and eventually overcome the stressful environment of low oxygen, growth factors, glucose and pH in the growing tumor mass. The requirement for neoangiogenesis to support tumor growth is now well established and is the basis for several promising anti-tumor modalities. Based on published data from our labs and others, we propose that the Unfolded Protein Response also plays a crucial role in adaptation to hypoxic stress, and like angiogenesis, represents an "encompassing and stable aspect of tumor development" and thus provides a unique opportunity for therapeutic exploitation. This proposal aims to identify agents that target key components of this adaptive response has the potential to offer additional and novel approaches to target the very stresses that hinder existing anti-tumor treatments and thereby improve antitumor treatment efficacy.

描述（由申请人提供）：缺氧/缺氧是大多数实体瘤微环境中特征良好的组成部分。大量的实验和临床证据支持了这样的观念，即缺氧从根本上改变了肿瘤的生理学对更具侵略性的表型。展开的蛋白质反应（UPR）是一种细胞应激适应途径，在存在内质网应激的情况下促进细胞存活，包括肿瘤微环境中的生理应激。 PERK/EIF2A/ATF4途径降低了蛋白质翻译的全球速率，从而减轻了缺氧下的代谢应激，同时诱导了重要的伴侣的转化上调和促生寿命基因。另一个UPR组件是激活核酸内切酶/激酶IRE1及其直接效应子XBP1，它激活了旨在增加ER折叠能力的转录程序。来自两个Co-PI的实验室的最新体外和体内研究表明，与来自Intact UPR的细胞相比，在体外变化的UPR反应的转化细胞表现出降低对缺氧的耐受性，并形成肿瘤较慢，表明UPR激活有助于肿瘤。在初步研究中，我们开发了基于细胞和动物的基于细胞的测定，用于UPR激活及其抑制小分子。该提案的总体目标是验证UPR激活为重要的抗肿瘤靶标，并使用新型的体外和体内测定方法来鉴定这种反应的有效抑制剂作为新型化学治疗剂。在AIM 1中，我们将采用PERK活性的体外报告基因测定法，以鉴定PERK/EIF21/ATF4途径的特定抑制剂，并测试这些抑制剂与IRE-1途径抑制剂（IRESTATINS（IRESTATIS）（IRESTATINS）在Normoxia和低氧下肿瘤生存的影响。在AIM 2中，我们将研究PERK和IRE1途径抑制剂与蛋白酶体抑制剂bortezomib的潜在协同作用，以在体外和体内杀死低氧肿瘤细胞。 AIM 3的研究将评估斑马鱼作为模型系统的使用，以分析PERK和IRE1途径抑制剂抑制异种移植的人类肿瘤并抑制血管生成而不引起显着发育异常的能力。最后，在AIM 4中，我们将测试小鼠肿瘤模型中已鉴定化合物的功效和潜在毒性。我们预计，这些努力将在单独的UPR的特定和有效抑制剂的开发中达到顶峰，或与现有的抗肿瘤剂和方式结合使用，将有效地减少临床前和临床恶性肿瘤的肿瘤负担。公共卫生相关性：实体瘤的标志是必须适应低氧，生长因子，葡萄糖和pH值的压力环境，并最终克服了肿瘤质量的压力。新血管生成支持肿瘤生长的需求现在已经建立得很好，并且是几种有前途的抗肿瘤方式的基础。基于我们实验室和其他人的公开数据，我们建议展开的蛋白质反应在适应低氧应激中也起着至关重要的作用，并且像血管生成一样，代表了“肿瘤发育的涵盖和稳定方面”，因此为治疗剥削提供了独特的机会。该建议旨在确定针对这种自适应反应的关键组成部分的药物，有可能提供其他新颖的方法来针对阻碍现有抗肿瘤治疗的压力，从而提高抗肿瘤治疗效果。