Canada's Michael Smith Genome Sciences Centre, BC Cancer Research Institute, Vancouver, British Columbia, Canada.
Department of Molecular Oncology, BC Cancer Research Institute, Vancouver, British Columbia, Canada.
Clin Cancer Res. 2024 Mar 1;30(5):1022-1037. doi: 10.1158/1078-0432.CCR-23-2187.
Ewing sarcoma is the second most common bone sarcoma in children, with 1 case per 1.5 million in the United States. Although the survival rate of patients diagnosed with localized disease is approximately 70%, this decreases to approximately 30% for patients with metastatic disease and only approximately 10% for treatment-refractory disease, which have not changed for decades. Therefore, new therapeutic strategies are urgently needed for metastatic and refractory Ewing sarcoma.
This study analyzed 19 unique Ewing sarcoma patient- or cell line-derived xenografts (from 14 primary and 5 metastatic specimens) using proteomics to identify surface proteins for potential immunotherapeutic targeting. Plasma membranes were enriched using density gradient ultracentrifugation and compared with a reference standard of 12 immortalized non-Ewing sarcoma cell lines prepared in a similar manner. In parallel, global proteome analysis was carried out on each model to complement the surfaceome data. All models were analyzed by Tandem Mass Tags-based mass spectrometry to quantify identified proteins.
The surfaceome and global proteome analyses identified 1,131 and 1,030 annotated surface proteins, respectively. Among surface proteins identified, both approaches identified known Ewing sarcoma-associated proteins, including IL1RAP, CD99, STEAP1, and ADGRG2, and many new cell surface targets, including ENPP1 and CDH11. Robust staining of ENPP1 was demonstrated in Ewing sarcoma tumors compared with other childhood sarcomas and normal tissues.
Our comprehensive proteomic characterization of the Ewing sarcoma surfaceome provides a rich resource of surface-expressed proteins in Ewing sarcoma. This dataset provides the preclinical justification for exploration of targets such as ENPP1 for potential immunotherapeutic application in Ewing sarcoma. See related commentary by Bailey, p. 934.
尤文肉瘤是儿童中第二常见的骨肉瘤，在美国每 150 万人中就有 1 例。尽管诊断为局限性疾病的患者的存活率约为 70%，但转移性疾病患者的存活率下降至约 30%，治疗耐药性疾病患者的存活率仅约为 10%，几十年来这些数据并未改变。因此，转移性和耐药性尤文肉瘤迫切需要新的治疗策略。
本研究通过蛋白质组学分析了 19 个独特的尤文肉瘤患者或细胞系衍生的异种移植物（来自 14 个原发性和 5 个转移性标本），以鉴定潜在免疫治疗靶点的表面蛋白。使用密度梯度超速离心法富集质膜，并与以类似方式制备的 12 个永生化非尤文肉瘤细胞系的参考标准进行比较。同时，对每个模型进行了全局蛋白质组分析，以补充表面组数据。所有模型均通过基于串联质量标签的质谱分析进行分析，以定量鉴定的蛋白质。
表面组和全局蛋白质组分析分别鉴定了 1,131 个和 1,030 个注释的表面蛋白。在鉴定的表面蛋白中，这两种方法都鉴定了已知的尤文肉瘤相关蛋白，包括 IL1RAP、CD99、STEAP1 和 ADGRG2，以及许多新的细胞表面靶标，包括 ENPP1 和 CDH11。与其他儿童肉瘤和正常组织相比，在尤文肉瘤肿瘤中证实了 ENPP1 的强染色。
我们对尤文肉瘤表面组的全面蛋白质组学特征描述提供了尤文肉瘤中丰富的表面表达蛋白资源。该数据集为探索 ENPP1 等靶标在尤文肉瘤中潜在免疫治疗应用的临床前依据提供了支持。请参阅 Bailey 的相关评论，第 934 页。
