Miljkovic-Licina Marijana, Arraud Nicolas, Zahra Aicha Dorra, Ropraz Patricia, Matthes Thomas
Laboratory for R&D in Hematology, Center for Translational Research in Onco-Hematology, University of Geneva Medical School, 1206 Geneva, Switzerland.
Department of Oncology, Hematology Service, Geneva University Hospitals, 1205 Geneva, Switzerland.
Cancers (Basel). 2021 Dec 23;14(1):56. doi: 10.3390/cancers14010056.
Extracellular vesicles (EVs) act in cell-to-cell communication, delivering cargo from donor to recipient cells and modulating their physiological condition. EVs secreted by leukemic blasts in patients with leukemia have been shown to influence the fate of recipient cells in the bone marrow microenvironment. Methods to quantify and to characterize them phenotypically are therefore urgently needed to study their functional role in leukemia development and to evaluate their potential as targets for therapy. We have used cryo-electron microscopy to study morphology and size of leukemic EVs, and nanoparticle tracking analysis and fluorescence triggering flow cytometry to quantify EVs in platelet-free plasma from a small cohort of leukemia patients and healthy blood donors. Additional studies with a capture bead-based assay allowed us to establish phenotypic signatures of leukemic EVs from 17 AML and 3 B-ALL patients by evaluating the expression of 37 surface antigens. In addition to tetraspanins and lineage-specific markers we found several adhesion molecules (CD29, and CD146) to be highly expressed by EVs from B-ALL and several leukemic stem cell antigens (CD44, CD105, CD133, and SSEA-4) to be expressed by EVs from AML patients. Further improvements in analytical methods to study EVs are needed before potentially using them as biomarkers for leukemia prognosis and follow-up.
细胞外囊泡（EVs）参与细胞间通讯，将物质从供体细胞传递至受体细胞并调节其生理状态。白血病患者白血病原始细胞分泌的EVs已被证明会影响骨髓微环境中受体细胞的命运。因此，迫切需要量化和表型特征化EVs的方法，以研究它们在白血病发展中的功能作用，并评估它们作为治疗靶点的潜力。我们利用冷冻电子显微镜研究白血病EVs的形态和大小，并使用纳米颗粒跟踪分析和荧光触发流式细胞术对一小群白血病患者和健康献血者的无血小板血浆中的EVs进行量化。通过基于捕获珠的检测进行的额外研究，使我们能够通过评估37种表面抗原的表达，建立17例急性髓系白血病（AML）和3例B淋巴细胞白血病（B-ALL）患者白血病EVs的表型特征。除了四跨膜蛋白和谱系特异性标志物外，我们发现几种黏附分子（CD29和CD146）在B-ALL来源的EVs中高表达，几种白血病干细胞抗原（CD44、CD105、CD133和阶段特异性胚胎抗原-4）在AML患者来源的EVs中表达。在潜在地将EVs用作白血病预后和随访的生物标志物之前，还需要进一步改进研究EVs的分析方法。
