Gene expression signature based screening in Ewing sarcoma

基于基因表达特征的尤文肉瘤筛查

• 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
• 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

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.

项目摘要 尤因肉瘤是由EWS-FLI1融合引起的高度侵略性骨骼和软组织癌 蛋白质。 EWS-FLI1癌蛋白功能部分作为异常转录因子，是必需的 肿瘤生长和生存。但是，直接用药物将EWS-FLI1靶向EWS-FLI1一直具有挑战性，作为一个 结果，迫切需要识别EWS-FLI1的下游目标和所产生的独特漏洞 通过癌蛋白。为了识别EWS-FLI1的下游靶标，我们使用了人类胚胎干细胞 表达可诱导的EWS-FLI1可以建模遗传学中的Ewing肉瘤的启动和开发 定义的系统。然后，我们使用此模型系统和一种基于基因表达的方法来识别该ewing 肉瘤细胞非常容易受到核糖核苷酸还原酶（RNR）的抑制剂，包括吉西他滨， 通过阻止脱氧核糖核苷酸的合成来损害DNA复制。而且，我们也 鉴定出RNR在尤因肉瘤细胞中的抑制作用会导致复制应力，激活展开 蛋白质反应，以及在响应DNA响应所需的蛋白质合成中的块 复制。值得注意的是，吉西他滨与检查点激酶1（CHK1）的抑制剂的组合（主要） 调节对DNA复制受损的响应的调节剂，导致体外协同作用和显着的延长 异种移植实验中的小鼠存活。在强大的初步数据的指导下，我们现在将追求 以下特定目的：1）剖析展开蛋白的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因肉瘤细胞的反应； 2）测试体内功效 使用EWING肉瘤异种移植模型，吉西他滨和第二代CHK1抑制剂Prexasertib的摄氏； 3）使用高通量筛查测试使用我们的基因表达鉴定的候选疗法 基于对EWING肉瘤细胞活性的方法。这些目标将通过结合 在癌细胞系，细胞系异种移植物，患者衍生的异种移植物以及我们的等源性的遗传学上的方法 定义的细胞系。这项工作将是重要的，因为预计它将与 治疗儿童和成人患有尤因肉瘤肿瘤，并更广泛地了解 尤因肉瘤肿瘤中肿瘤发生的基本机制。拟议的研究具有创新性，因为它 将EWING肉瘤的新型干细胞模型与基因表达签名筛选相结合 识别新的治疗靶标的方法。