Lovisa Federica, Garbin Anna, Crotti Sara, Di Battista Piero, Gallingani Ilaria, Damanti Carlotta Caterina, Tosato Anna, Carraro Elisa, Pillon Marta, Mafakheri Erfan, Romanato Filippo, Gaffo Enrico, Biffi Alessandra, Bortoluzzi Stefania, Agostini Marco, Mussolin Lara
Maternal and Child Health Department, Padova University, 35128 Padova, Italy.
Istituto di Ricerca Pediatrica Città della Speranza, 35127 Padova, Italy.
Diagnostics (Basel). 2021 Feb 6;11(2):253. doi: 10.3390/diagnostics11020253.
Over the past 15 years, several biological and pathological characteristics proved their significance in pediatric anaplastic lymphoma kinase (ALK)-positive anaplastic large-cell lymphoma (ALCL) prognostic stratification. However, the identification of new non-invasive disease biomarkers, relying on the most important disease mechanisms, is still necessary. In recent years, plasmatic circulating small extracellular vesicles (S-EVs) gathered great importance both as stable biomarker carriers and active players in tumorigenesis. In the present work, we performed a comprehensive study on the proteomic composition of plasmatic S-EVs of pediatric ALCL patients compared to healthy donors (HDs). By using a mass spectrometry-based proteomics approach, we identified 50 proteins significantly overrepresented in S-EVs of ALCL patients. Gene Ontology enrichment analysis disclosed cellular components and molecular functions connected with S-EV origin and vesicular trafficking, whereas cell adhesion, glycosaminoglycan metabolic process, extracellular matrix organization, collagen fibril organization and acute phase response were the most enriched biological processes. Of importance, consistently with the presence of nucleophosmin (NPM)-ALK fusion protein in ALCL cells, a topological enrichment analysis based on Reactome- and Kyoto Encyclopedia of Genes and Genomes (KEGG)-derived networks highlighted a dramatic increase in proteins of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway in ALCL S-EVs, which included heat shock protein 90-kDa isoform alpha 1 (HSP90AA1), osteopontin (SPP1/OPN) and tenascin C (TNC). These results were validated by Western blotting analysis on a panel of ALCL and HD cases. Further research is warranted to better define the role of these S-EV proteins as diagnostic and, possibly, prognostic parameters at diagnosis and for ALCL disease monitoring.
在过去15年中，多种生物学和病理学特征在儿童间变性淋巴瘤激酶（ALK）阳性间变性大细胞淋巴瘤（ALCL）的预后分层中显示出重要意义。然而，基于最重要的疾病机制鉴定新的非侵入性疾病生物标志物仍然很有必要。近年来，血浆循环小细胞外囊泡（S-EV）作为稳定的生物标志物载体和肿瘤发生中的活跃参与者变得极为重要。在本研究中，我们对儿童ALCL患者与健康供者（HD）血浆S-EV的蛋白质组组成进行了全面研究。通过基于质谱的蛋白质组学方法，我们鉴定出50种在ALCL患者的S-EV中显著富集的蛋白质。基因本体富集分析揭示了与S-EV起源和囊泡运输相关的细胞成分和分子功能，而细胞黏附、糖胺聚糖代谢过程、细胞外基质组织、胶原纤维组织和急性期反应是最富集的生物学过程。重要的是，与ALCL细胞中存在核磷蛋白（NPM）-ALK融合蛋白一致，基于Reactome和京都基因与基因组百科全书（KEGG）衍生网络的拓扑富集分析突出显示，ALCL S-EV中磷脂酰肌醇3激酶（PI3K）/AKT通路的蛋白质显著增加，其中包括热休克蛋白90-kDa异构体α1（HSP90AA1）、骨桥蛋白（SPP1/OPN）和腱生蛋白C（TNC）。这些结果通过对一组ALCL和HD病例的蛋白质免疫印迹分析得到验证。有必要进一步研究以更好地确定这些S-EV蛋白作为诊断标志物以及可能作为诊断时和ALCL疾病监测的预后参数的作用。
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