Mass spectrometry-based proteomics analysis of whole cell and secreted extracellular vesicles of four diffuse large B-cell lymphoma cell lines

• Autor(a) principal: Baeta, Henrique Silva Caio Simões
• Data de Publicação: 2021
• Tipo de documento: Dissertação
• Idioma: eng
• Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
• Texto Completo: http://hdl.handle.net/10362/118632
• topic: B-cell Lymphoma; Cell Lines; Proteomics Analysis; Mass Spectrometry; Ciências Médicas

DISCUSSION: Here we implemented an experimental and computational workflow to study the protein expression profiles of small extracellular vesicles from in vitro cultured DLBCL cell lines. Our main question was whether the proteomes of small extracellular vesicles could be used to segregate the prognostic cell of origin subtypes of DLBCL. We selected 4 DLBCL cell line models, the DB and HT cells representing germinal center b cell like (GCB) and RIVA and OCI-ly3 cells as activated b cell like (ABC) DLBCLs. We cultured cells considering suppliers instructions to make sure our results are comparable to others using the same models. We tried to truthfully report our experimental conditions and sEVs purification protocol to prevent possible misinterpretation of data. To our knowledge this is the first high accuracy, global, quantitative proteomics study to comprehensively compare the proteomes of sEVs derived from DLBCL cells. We considered the expression of other biomolecules in sEVs. However, proteins are main effectors of encoding genes and among the most frequently therapeutically targeted biomolecules. We decided to take advantage of recent developments in mass spectrometry instruments147 and data analysis strategies127 to investigate DLBCL sEVs proteomes.