Discovering the infectome of human endothelial cells challenged with Aspergillus fumigatus applying a mass spectrometry label-free approach

• Published in: Journal of proteomics 2014-01, Vol.97, p.126-140
• Main Authors: Curty, N., Kubitschek-Barreira, P.H., Neves, G.W., Gomes, D., Pizzatti, L., Abdelhay, E., Souza, G.H.M.F., Lopes-Bezerra, L.M.
• Format: Article
• Language: English
• Subjects: A. fumigatus; Aspergillus fumigatus; Endothelial cell; HUVEC; Infectome; Invasive aspergillosis; NanoLC-HDMSE
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2013.07.003

Blood vessel invasion is a key feature of invasive aspergillosis. This angioinvasion process contributes to tissue thrombosis, which can impair the access of leukocytes and antifungal drugs to the site of infection. It has been demonstrated that human umbilical vein endothelial cells (HUVECs) are activated and assume a prothrombotic phenotype following contact with Aspergillus fumigatus hyphae or germlings, a process that is independent of fungus viability. However, the molecular mechanisms by which this pathogen can activate endothelial cells, together with the endothelial pathways that are involved in this process, remain unknown. Using a label-free approach by High Definition Mass Spectrometry (HDMSE), differentially expressed proteins were identified during HUVEC–A. fumigatus interaction. Among these, 89 proteins were determined to be up- or down-regulated, and another 409 proteins were exclusive to one experimental condition: the HUVEC control or HUVEC:AF interaction. The in silico predictions provided a general view of which biological processes and/or pathways were regulated during HUVEC:AF interaction, and they mainly included cell signaling, immune response and hemostasis pathways. This work describes the first global proteomic analysis of HUVECs following interaction with A. fumigatus germlings, the fungus morphotype that represents the first step of invasion and dissemination within the host. A. fumigatus causes the main opportunistic invasive fungal infection related to neutropenic hematologic patients. One of the key steps during the establishment of invasive aspergillosis is angioinvasion but the mechanism associated with the interaction of A. fumigatus with the vascular endothelium remains unknown. The identification of up- and down-regulated proteins expressed by human endothelial cells in response to the fungus infection can contribute to reveal the mechanism of endothelial response and, to understand the physiopathology of this high mortality disease. This article is part of a Special Issue entitled: Trends in Microbial Proteomics. [Display omitted] •A nanoUPLC-HDMSE was used to study the proteome of infected HUVECs by A. fumigatus.•In silico analysis of proteins up- or down-regulated in the HUVEC:AF model is shown.•In silico analysis of proteins exclusively expressed in the HUVEC:AF condition.•Protein interaction map showed possible routes involved in the endothelial response.•This work describes proteins implicated in immune response an