Author(s): Ejtehadifar, Mostafa ; Zahedi, Sara ; Gameiro, Paula ; Cabeçadas, José ; Rodriguez, Manuel S. ; da Silva, Maria Gomes ; Beck, Hans Christian ; Matthiesen, Rune ; Carvalho, Ana Sofia
Date: 2025
Persistent ID: http://hdl.handle.net/10362/182594
Origin: Repositório Institucional da UNL
Subject(s): Diffuse large B cell lymphoma; Extracellular vesicles; Peripheral immune cells; Polyubiquitination; Proteomics; Molecular Medicine; Molecular Biology; SDG 3 - Good Health and Well-being
Funding Information: R.M. is supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia (CEEC position, DOI:10.54499/CEECIND/03906/2017/CP1421/CT0004 ). A.S.C. is supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia ( DOI:10.54499/DL57/2016/CP1457/CT0013 ). This work is funded by FEDER funds through the COMPETE 2020 Programme and National Funds through FCT \u2014Portuguese Foundation for Science and Technology under the projects number PTDC/BTM-TEC/1746/2021 , PTDC/BTM-TEC/30087/2017 and PTDC/BTM-TEC/30088/2017 . The project was supported by from European Union to advance EV research ( Horizon2020_GA_n\u00B0101079264 , EVCA). We acknowledge the COST Action CA20113 \u201CPROTEOCURE\u201D supported by COST ( European Cooperation in Science and Technology ). This article is also a result of the projects ( iNOVA4Health\u2013UIDB/04462/2020 and UIDP/04462/2020 ), and by the Associated Laboratory LS4FUTURE ( LA/P/0087/2020 ), two programs financially supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia / Minist\u00E9rio da Ci\u00EAncia, Tecnologia e Ensino Superior . We thank Instituto Gulbenkian de Ci\u00EAncia for use of transmission electron microscopy. Funding Information: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Ana Sofia Carvalho and Rune Matthiesen reports financial support, article publishing charges, and equipment, drugs, or supplies were provided by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia. Rune Matthiesen reports financial support was provided by Horizon 2020. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.R.M. is supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia (CEEC position, DOI:10.54499/CEECIND/03906/2017/CP1421/CT0004). A.S.C. is supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia (DOI:10.54499/DL57/2016/CP1457/CT0013). This work is funded by FEDER funds through the COMPETE 2020 Programme and National Funds through FCT\u2014Portuguese Foundation for Science and Technology under the projects number PTDC/BTM-TEC/1746/2021, PTDC/BTM-TEC/30087/2017 and PTDC/BTM-TEC/30088/2017. The project was supported by from European Union to advance EV research (Horizon2020_GA_n\u00B0101079264, EVCA and Horizon Europe-SE2023/GA101183034). We acknowledge the COST Action CA20113 \u201CPROTEOCURE\u201D supported by COST (European Cooperation in Science and Technology). This article is also a result of the projects (iNOVA4Health\u2013UIDB/04462/2020 and UIDP/04462/2020), and by the Associated Laboratory LS4FUTURE (LA/P/0087/2020), two programs financially supported by Funda\u00E7\u00E3o para a Ci\u00EAncia e a Tecnologia/Minist\u00E9rio da Ci\u00EAncia, Tecnologia e Ensino Superior. We thank Instituto Gulbenkian de Ci\u00EAncia for use of transmission electron microscopy. Publisher Copyright: © 2025 The Authors
Diffuse large B-cell lymphoma (DLBCL) is an aggressive disease and a frequent form of non-Hodgkin lymphoma. Given the primary localization of DLBCL and the effect of tumors on the systemic immune response, we investigated the proteome of DLBCL patients' and healthy donors (HDs') peripheral immune cells (PICs). Since the ubiquitin-proteasome system has a vital role in proteome regulation and immune cells' functions, this study also explores the potential impact of DLBCL secretome on the polyubiquitination level in PICs. PICs from DLBCL patients and HDs were isolated and analyzed by mass spectrometry-based proteomics. The analysis resulted in 135 down and 51 upregulated proteins (adjusted p-value <0.05). Unsupervised principal component analysis revealed distinct proteomic profiles between DLBCL and HDs. Functional enrichment analysis for comparison between DLBCL and HDs-PICs proteome identified immune-related pathways such as innate immune system, specifically neutrophil degranulation, Fcγ receptor-dependent phagocytosis, and JAK-STAT signaling after IL-12 stimulation as downregulated. Proteomics analysis of DLBCL-PICs also showed dysregulation of proteostasis factors. This prompted the investigation of the effect of tumor secretome on viability and polyubiquitination level in mononuclear immune cells. Therefore, human HD peripheral blood mononuclear cells (PBMCs) were cultured in the presence of DLBCL cell line-derived soluble factors, small-EVs, and large-EVs in vitro. Our results revealed that exposure of mainly small-EVs, and large-EVs to HD PBMCs increased the polyubiquitination in PBMCs and decreased PIC viability. These findings suggest impaired immune responses in DLBCL-PICs, with tumor secretome-inducing polyubiquitination and reduced PIC viability.
