Increased Tenascin C, Osteopontin and HSP90 Levels in Plasmatic Small Extracellular Vesicles of Pediatric ALK-Positive Anaplastic Large Cell Lymphoma: New Prognostic Biomarkers?

Abstract

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.

Evaluating Differences in the Proteome Within Extracellular Vesicles Between Pre- Malignant and Malignant Plasma Cell Disorders

“…12 Recently, SEV-derived SAA levels have also been found to be increased in pediatric patients with anaplastic lymphoma kinase (ALK)-positive anaplastic large-cell lymphoma (ALCL) relative to healthy controls. 13 Therefore, increased SAA levels in SEVs may also re ect the increased disease burden of malignant PCs and/or an in ammatory state within the bone marrow.…”

Extracellular Vesicles for Childhood Cancer Liquid Biopsy

“…Lovisa et al [97] performed a proteomics study to compare plasma EV derived from pediatric ALCL patients and healthy donors and identified up to 50 proteins that were significant in EVs derived from ALCL patients [97]. Functional enrichment analysis revealed that these proteins are related to cell adhesion, glycosaminoglycan metabolic process, extracellular matrix organization, collagen fibril organization, and acute phase response, as well as the PI3K/AKT pathway [97].…”

“…Functional enrichment analysis revealed that these proteins are related to cell adhesion, glycosaminoglycan metabolic process, extracellular matrix organization, collagen fibril organization, and acute phase response, as well as the PI3K/AKT pathway [97]. This study suggested that these ALCL-derived EV proteins can be used as diagnostic and possibly prognostic parameters during diagnosis and ALCL disease monitoring [97]. Another proteomic study identified 11 unique proteins, including five upregulated in non-relapsed HL (e.g., isoform 2 preproprotein of complement C4-A, complement C4-B, fibrinogen γ chain, inter-α-trypsin inhibitor heavy chain H2, and immunoglobulin heavy chain constant region mu) and six upregulated in relapsed HL (e.g., apolipoprotein A-I, apolipoprotein A-IV, clusterin, haptoglobin, α-1-acid glycoprotein 1, and transthyretin) [98].…”

New Advances in Liquid Biopsy Technologies for Anaplastic Lymphoma Kinase (ALK)—Positive Cancer

“…A parallel proteomic analysis led to the identification of proteins involved in PI3K signaling that are enriched in exosomes from ALCL patients. The authors proposed three proteins, namely tenascin C, osteopontin and heat shock protein 90 as potential biomarkers for ALCL prognostic stratification [74]. Altogether, these studies open the possibility to assess the risk of relapse and to monitor the response to therapy in a disease where tissue re-biopsies are often difficult to obtain.…”