Gene expression signature based screening in Ewing sarcoma

基于基因表达特征的尤文肉瘤筛查

基本信息

批准号：
10329929
负责人：
David J Gordon
金额：
$ 37.38万
依托单位：
UNIVERSITY OF IOWA
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：
2018-02-02 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10329929
关键词：
Adolescent Adult Biological Models Bone Tissue CHEK1 gene Cancer cell line Cell Line Cell model Cells Child Chromosomal translocation Clinical Data Clinical Trials Cytotoxic Chemotherapy DNA Damage DNA biosynthesis Data Deoxyribonucleotides Development Disease Drug Screening EWS-FLI1 fusion protein EWSR1 gene Ewings sarcoma FDA approved FLI1 gene Gene Expression Gene Expression Profile Gene Fusion Generations Genetic Goals Immune checkpoint inhibitor Impairment In Vitro Laboratories Lead Libraries Malignant Childhood Neoplasm Malignant Neoplasms Mammalian Cell Mediator of activation protein Modeling Molecular Molecular Target Mus Network-based Oncogenes Oncoproteins Operative Surgical Procedures Pathogenesis Pathway interactions Pharmaceutical Preparations Pharmacology Protein Biosynthesis Proteins Radiation Regulation Research Resources Ribonucleotide Reductase Ribonucleotide Reductase Inhibitor Role Soft tissue sarcoma System Testing Toxic effect United States National Institutes of Health Work Xenograft Model Xenograft procedure base bone chemotherapy clinical translation cytotoxic drug candidate drug sensitivity early phase clinical trial experimental study gemcitabine high throughput screening human disease human embryonic stem cell in vivo in vivo evaluation inhibitor innovation mouse model neoplastic cell new therapeutic target novel patient derived xenograft model pre-clinical replication stress response screening soft tissue stem cell model subcutaneous synergism targeted treatment therapeutic candidate therapeutic target transcription factor tumor tumor growth tumor initiation tumor progression tumorigenesis

项目摘要

PROJECT SUMMARY Ewing sarcoma is a highly aggressive bone and soft tissue cancer that is caused by the EWS-FLI1 fusion protein. The EWS-FLI1 oncoprotein functions, in part, as an aberrant transcription factor and is required for tumor growth and survival. However, directly targeting EWS-FLI1 with drugs has been challenging and, as a result, there is a critical need to identify downstream targets of EWS-FLI1 and unique vulnerabilities incurred by the oncoprotein. In order to identify downstream targets of EWS-FLI1, we used human embryonic stem cells that express inducible EWS-FLI1 to model the initiation and development of Ewing sarcoma in a genetically defined system. We then used this model system and a gene expression based approach to identify that Ewing sarcoma cells are uniquely vulnerable to inhibitors of ribonucleotide reductase (RNR), including gemcitabine, which impair DNA replication by blocking the synthesis of deoxyribonucleotides. Moreover, we have also identified that the inhibition of RNR in Ewing sarcoma cells results replication stress, activation of the unfolded protein response, and a block in the synthesis of proteins required for the response to impaired DNA replication. Notably, the combination of gemcitabine with an inhibitor of checkpoint kinase 1 (CHK1), the major regulator of the response to impaired DNA replication, results in synergy in vitro and a significant prolongation of mouse survival in xenograft experiments. Guided by strong preliminary data, we will now pursue the following specific aims: 1) dissect the molecular basis of the activation and regulation of the unfolded protein response in Ewing sarcoma cells treated with inhibitors of ribonucleotide reductase; 2) test the in vivo efficacy of gemcitabine and a second-generation CHK1 inhibitor, prexasertib, using Ewing sarcoma xenograft models; and 3) use high-throughput screening to test candidate therapeutics identified using our gene expression based approach for activity against Ewing sarcoma cells. These aims will be tested using a combination of approaches in cancer cell lines, cell line xenografts, patient-derived xenografts, and our isogenic, genetically defined cell lines. This work will be significant because it is expected to have translational relevance for the treatment of children and adults with Ewing sarcoma tumors, as well as lead to a broader understanding of the basic mechanisms of tumorigenesis in Ewing sarcoma tumors. The proposed research is innovative because it integrates a novel, stem cell model of Ewing sarcoma with a gene expression signature based screening approach to identify new therapeutic targets.

项目概要尤文肉瘤是一种高度侵袭性的骨和软组织癌症，由 EWS-FLI1 融合引起蛋白质。 EWS-FLI1 癌蛋白的部分功能是作为异常转录因子，并且是肿瘤生长和存活。然而，用药物直接靶向 EWS-FLI1 一直具有挑战性，并且因此，迫切需要识别 EWS-FLI1 的下游目标和所产生的独特漏洞由癌蛋白。为了确定 EWS-FLI1 的下游靶标，我们使用了人胚胎干细胞表达诱导型 EWS-FLI1 来模拟尤文肉瘤的发生和发展定义的系统。然后我们使用这个模型系统和基于基因表达的方法来识别尤因肉瘤细胞特别容易受到核糖核苷酸还原酶 (RNR) 抑制剂的影响，包括吉西他滨、它通过阻断脱氧核糖核苷酸的合成来损害DNA复制。此外，我们还有发现尤文肉瘤细胞中 RNR 的抑制会导致复制应激、未折叠细胞的激活蛋白质反应，以及对受损 DNA 做出反应所需的蛋白质合成受阻复制。值得注意的是，吉西他滨与检查点激酶 1 (CHK1) 抑制剂的组合是主要的 DNA复制受损反应的调节剂，在体外产生协同作用并显着延长异种移植实验中小鼠的存活率。在强有力的初步数据的指导下，我们现在将追求以下具体目标：1）剖析未折叠蛋白激活和调节的分子基础用核糖核苷酸还原酶抑制剂处理的尤文肉瘤细胞的反应； 2）体内功效测试使用尤文肉瘤异种移植模型研究吉西他滨和第二代 CHK1 抑制剂 prexasertib； 3) 使用高通量筛选来测试使用我们的基因表达确定的候选疗法基于针对尤文肉瘤细胞活性的方法。这些目标将使用以下组合进行测试癌细胞系、细胞系异种移植物、患者来源的异种移植物以及我们的同基因、遗传性方法定义的细胞系。这项工作意义重大，因为预计它将对以下领域具有转化意义：治疗患有尤文肉瘤的儿童和成人，并导致对尤因肉瘤肿瘤的更广泛了解尤文肉瘤肿瘤发生的基本机制。拟议的研究具有创新性，因为它将尤文肉瘤的新型干细胞模型与基于基因表达特征的筛选相结合方法来确定新的治疗靶点。