Culture of <i>Mycobacterium smegmatis</i> in Different Carbon Sources to Induce In Vitro Cholesterol Consumption Leads to Alterations in the Host Cells after Infection: A Macrophage Proteomics Analysis
Jaqueline Batista de Lima,
Lana Patricia da Silva Fonseca,
Luciana Pereira Xavier,
Barbarella de Matos Macchi,
Juliana Silva Cassoli,
Edilene Oliveira da Silva,
Rafael Borges da Silva Valadares,
José Luiz Martins do Nascimento,
Agenor Valadares Santos,
Chubert Bernardo Castro de Sena
Affiliations
Jaqueline Batista de Lima
Laboratory of Structural Biology, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Lana Patricia da Silva Fonseca
Instituto Tecnológico Vale, Belém 66055-090, PA, Brazil
Luciana Pereira Xavier
Laboratory of Biotechnology of Enzymes and Biotransformations, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Barbarella de Matos Macchi
Laboratory of Molecular and Cellular Neurochemistry, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Juliana Silva Cassoli
Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Edilene Oliveira da Silva
Laboratory of Structural Biology, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Rafael Borges da Silva Valadares
Instituto Tecnológico Vale, Belém 66055-090, PA, Brazil
José Luiz Martins do Nascimento
Laboratory of Molecular and Cellular Neurochemistry, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Agenor Valadares Santos
Laboratory of Biotechnology of Enzymes and Biotransformations, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
Chubert Bernardo Castro de Sena
Laboratory of Structural Biology, Institute of Biological Sciences, Federal University of Pará, Belém 66075-110, PA, Brazil
During tuberculosis, Mycobacterium uses host macrophage cholesterol as a carbon and energy source. To mimic these conditions, Mycobacterium smegmatis can be cultured in minimal medium (MM) to induce cholesterol consumption in vitro. During cultivation, M. smegmatis consumes MM cholesterol and changes the accumulation of cell wall compounds, such as PIMs, LM, and LAM, which plays an important role in its pathogenicity. These changes lead to cell surface hydrophobicity modifications and H2O2 susceptibility. Furthermore, when M. smegmatis infects J774A.1 macrophages, it induces granuloma-like structure formation. The present study aims to assess macrophage molecular disturbances caused by M. smegmatis after cholesterol consumption, using proteomics analyses. Proteins that showed changes in expression levels were analyzed in silico using OmicsBox and String analysis to investigate the canonical pathways and functional networks involved in infection. Our results demonstrate that, after cholesterol consumption, M. smegmatis can induce deregulation of protein expression in macrophages. Many of these proteins are related to cytoskeleton remodeling, immune response, the ubiquitination pathway, mRNA processing, and immunometabolism. The identification of these proteins sheds light on the biochemical pathways involved in the mechanisms of action of mycobacteria infection, and may suggest novel protein targets for the development of new and improved treatments.
