Characterization and targeting the CXCR2-STAT-1/3 axis in metastatic Ewing sarcoma

项目摘要 尤文肉瘤(EWS)是儿童第二常见的骨肿瘤。确诊时，大约25%-30% 的EWS患者有转移性疾病。对于局部疾病，全身化疗方案与 手术和/或放射治疗已将患者的存活率显著提高到约70%。 不幸的是，转移性EWS的预后仍然很差，5年存活率为20%-30%。 在过去的几十年里，这种预测没有改变。因此，当务之急是要重点研究 EWS转移，并确定转移肿瘤的特定生物学特征。在这方面，重要的是 研究肿瘤生物学不是孤立的，而是在肿瘤间质的背景下进行研究，以更好地理解相互作用 在肉瘤细胞和肿瘤相关基质细胞之间，促进了EWS的转移生长。vt.给出 典型脊椎动物EWS模型的局限性，我们整合肿瘤内在和间质的方法 导致转移表型的因素比研究肿瘤更接近生理背景 与世隔绝。利用系列移植方法，我们已经产生了新的肺转移EWS细胞系。 这些细胞与肺成纤维细胞共培养的细胞因子分析显示，几种 细胞因子，包括CXCL-1/2、CXCL-5和CXCL-8，它们是受体CXCR2的配体。这 利用纳米串技术进行转录分析证实了从配对的初级和 肺转移患者样本。此外，对来自多个转移模型的蛋白质组学进行综合分析， 在转移样本中以显著差异表达蛋白的形式聚集在STAT1上。因此，通过 多组学方法在相关创新体内转移模型和人类患者中的集成 样本，我们已经确定了一种新的EWS转移的分子机制，我们可以很容易地评估 利用我们独特的资源。我们的主要假设是CXCR2-STAT1轴是一条关键的途径 转移性EWS和一个有吸引力的治疗靶点。为了解决这一假设，并弥合目前的差距 在知识和潜在的治疗机会方面，我们提出了两个创新，翻译和探索性 目标。AIM1将表征CXCR2-STAT1通路在原发和转移中的致瘤作用 通过鉴定CXCR2诱导的STAT1调控基因并评估遗传学和药理学来研究肿瘤 抑制这一途径作为一种新的治疗干预措施。AIM2将研究细胞外的作用 囊泡作为肿瘤-间质沟通的细胞间介质，通过以下方式促进CXCR2-STAT1信号转导 微小RNA(MiRNA)调控。侵袭性和转移性尤文肉瘤的治疗仍然是 在治疗上挑战。我们的目标的成功完成将提供新的分子和有价值的预 转移性尤文肉瘤的临床治疗启示。

Project Summary Ewing Sarcoma (EwS) is the second most common bone tumor in children. At diagnosis, approximately 25-30% of patients with EwS have metastatic disease. For localized disease, systemic chemotherapy regimen along with surgery and/or radiotherapy has significantly increased the survival rate of patients to approximately 70%. Unfortunately, the prognosis for metastatic EwS has remained dismal, with a 5-year survival rate of 20–30%. This prognosis has not changed in last several decades. Thus, it is imperative to focus on the mechanisms of EwS metastasis and identify specific biological features of the metastatic tumor. In this regard, it is important to study the tumor biology not in isolation, but in the context of the tumor stroma, to better understand the interaction between the sarcoma cells and the tumor associated stromal cells that facilitates EwS metastatic growth. Given the limitation in representative vertebrate models for EwS, our approach of integrating tumor intrinsic and stromal factors that lead to metastatic phenotypes is closer to the physiological context than studying the tumor in isolation. Using serial transplantation approaches, we have generated novel lung metastatic EwS cell lines. Cytokine analysis from co-culture of these cells with lung fibroblast cells, revealed upregulation of several cytokines, including CXCL-1/2, CXCL-5, and CXCL-8, which are ligands to the receptor CXCR2. This observation was confirmed from transcriptomic analysis using Nanostring technology from paired primary with lung metastatic patient samples. Further, integrated analysis of proteomics from multiple metastatic models, converged on STAT1 as a significantly differentially expressed protein in metastatic samples. Thus, through the integration of multi-omic approaches on relevant innovative in vivo metastatic models and human patient specimens, we have identified a novel molecular mechanism of metastasis in EwS that we can readily assess using our unique resources. Our overarching hypothesis is that the CXCR2-STAT1 axis is a critical pathway in metastatic EwS and an attractive therapeutic target. To address this hypothesis, and to bridge the current gaps in knowledge and potential therapeutic opportunities, we propose two innovative, translation and exploratory aims. Aim1 will characterize the tumorigenic effects of the CXCR2-STAT1 pathway in primary and metastatic tumor by identifying STAT1 regulated genes induced by CXCR2 and assessing genetic and pharmacological inhibition of this pathway as a novel therapeutic intervention. Aim2 will investigate the role of extra cellular vesicles, as an intercellular mediator of tumor-stroma communication that facilitates CXCR2-STAT1 signaling by micro RNA (miRNA) regulation. The treatment of aggressive and metastatic Ewing sarcomas remains a challenge therapeutically. Successful completion of our aims will provide novel molecular, and valuable pre- clinical therapeutic insights for metastatic Ewing sarcoma.