Goodspeed Andrew, Bodlak Avery, Duffy Alexis B, Nelson-Taylor Sarah, Oike Naoki, Porfilio Timothy, Shirai Ryota, Walker Deandra, Treece Amanda, Black Jennifer, Donaldson Nathan, Cost Carrye, Garrington Timothy, Greffe Brian, Luna-Fineman Sandra, Demedis Jenna, Lake Jessica, Danis Etienne, Verneris Michael R, Adams Daniel L, Hayashi Masanori
University of Colorado Cancer Center, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
Department of Biomedical Informatics, University of Colorado Anschutz Medical Campus, Aurora, Colorado.
Clin Cancer Res. 2025 May 15;31(10):2010-2023. doi: 10.1158/1078-0432.CCR-24-2040.
Ewing sarcoma is the second most common bone cancer in children, accounting for 2% of pediatric cancer diagnoses. Patients who present with metastatic disease at the time of diagnosis have a dismal prognosis compared with the >70% 5-year survival of those with localized disease. Novel therapeutic approaches that can impact metastatic disease are desperately needed, as well as a deeper understanding of the heterogeneity of Ewing sarcoma tumors.
In this study, we utilized single-cell RNA sequencing to characterize the transcriptional landscape of primary Ewing sarcoma tumors and the surrounding tumor microenvironment in a cohort of seven untreated patients with Ewing sarcoma, as well as in circulating tumor cells (CTC). A potential CTC therapeutic target was evaluated through immunofluorescence of fixed CTCs from a separate cohort.
Primary tumor samples demonstrate a heterogeneous transcriptional landscape with several conserved gene expression programs, including those composed of genes related to proliferation and Ewing sarcoma gene targets, which were found to correlate with overall survival. Copy-number analysis identified subclonal evolution within patients prior to treatment. Analyses of the immune microenvironment reveal an immunosuppressive microenvironment with complex intercellular communication among the tumor and immune cells. Single-cell RNA sequencing and immunofluorescence of CTCs at the time of diagnosis identified TSPAN8 as a potential therapeutic target.
Ewing sarcoma tumors demonstrate significant transcriptional heterogeneity as well as a complex immunosuppressive microenvironment. This work evaluates several proposed targets that warrant further exploration as novel therapeutic strategies.
尤因肉瘤是儿童中第二常见的骨癌，占儿童癌症诊断病例的2%。与局限性疾病患者>70%的5年生存率相比，诊断时出现转移性疾病的患者预后较差。迫切需要能够影响转移性疾病的新型治疗方法，以及对尤因肉瘤肿瘤异质性的更深入理解。
在本研究中，我们利用单细胞RNA测序来表征一组7例未经治疗的尤因肉瘤患者的原发性尤因肉瘤肿瘤及其周围肿瘤微环境以及循环肿瘤细胞（CTC）的转录图谱。通过对来自另一队列的固定CTC进行免疫荧光分析，评估了一个潜在的CTC治疗靶点。
原发性肿瘤样本显示出具有几个保守基因表达程序的异质转录图谱，包括那些由与增殖相关的基因和尤因肉瘤基因靶点组成的程序，这些程序被发现与总生存期相关。拷贝数分析确定了治疗前患者体内的亚克隆进化。免疫微环境分析揭示了一种免疫抑制微环境，肿瘤细胞与免疫细胞之间存在复杂的细胞间通讯。诊断时对CTC进行的单细胞RNA测序和免疫荧光分析确定TSPAN8为一个潜在的治疗靶点。
尤因肉瘤肿瘤表现出显著的转录异质性以及复杂的免疫抑制微环境。这项工作评估了几个有望作为新型治疗策略进行进一步探索的靶点。
