Single cell analysis of kynurenine and System L amino acid transport in T cells

Sinclair, Linda V.; Neyens, Damien; Ramsay, George; Taylor, Peter M.; Cantrell, Doreen A.

doi:10.1038/s41467-018-04366-7

Cited by 136 publications

(119 citation statements)

References 36 publications

(49 reference statements)

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“…In corroboration with these data, we also observed increased amounts of transcripts related to the ASC survival or differentiation, such as TNFS13B and IL-10 ( Supplementary Figure 5) as previously described (30,(73)(74)(75). Thus, it is suggested that the tryptophan degradation would be the cause and not consequence of the DENV-mediated ASC differentiation in vitro, as it seems to occur earlier than the BLIMP1 expression (64). The role of this metabolic pathway in the massive ASC response detected in Dengue patients is still to be ascertained.…”

Section: Discussionsupporting

confidence: 90%

“…Genes with expressions tightly bound to the IDO activity, such as eukaryotic initiation factor 2 alpha (eIF2alpha) and kinase general control non-derepressible-2 (GCN2), can enhance the BLIMP1 (or PRDM1) expression in a tryptophan-depleted condition. On the other hand, the BLIMP1 protein binds to the IDO promoter, repressing its expression (64). The tryptophan degradation also modifies the expression of SLC7A5 (65) and WARS during this process in cancer cells (66) and ASCs (67).…”

Section: Discussionmentioning

confidence: 99%

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Flavivirus-Mediating B Cell Differentiation Into Antibody-Secreting Cells in Humans Is Associated With the Activation of the Tryptophan Metabolism

et al. 2020

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Patients infected with the Dengue virus (DENV) often present with a massive generation of DENV-specific antibody-secreting cells (ASCs) in the blood. In some cases, these ASCs represent more than 50% of the circulating B cells, a higher magnitude than those induced by other infections, vaccinations, and plasma cell lymphomas. However, it remains unclear how the DENV infection elicits this colossal response. To address this issue, we utilised an in vitro strategy to induce human PBMCs of healthy individuals incubated with DENV particles (DENV4 TVP/360) to differentiate into ASCs. As controls, PBMCs were incubated with a mitogen cocktail or supernatants of uninfected C6/36 cells (mock). The ASC phenotype and function were increasingly detected in the DENV and mitogen-cultured PBMCs as compared to mock-treated cells. In contrast to the in vivo condition, secreted IgG derived from the PBMC-DENV culture was not DENV-specific. Lower ASC numbers were observed when inactivated viral particles or purified B cells were added to the cultures. The physical contact was essential between B cells and the remaining PBMCs for the DENV-mediated ASC response. Considering the evidence for the activation of the tryptophan metabolism detected in the serum of Dengue patients, we assessed its relevance in the DENV-mediated ASC differentiation. For this, tryptophan and its respective metabolites were quantified in the supernatants of cell cultures through mass spectrophotometry. Tryptophan depletion and kynurenine accumulation were found in the supernatants of PBMC-DENV cultures, which presented enhanced detection of indoleamine 2,3-dioxygenase 1 and 2 transcripts as compared to controls. In PBMC-DENV cultures, tryptophan and kynurenine levels strongly correlated to the respective ASC numbers, while the kynurenine levels were directly proportional to the secreted IgG titers. Contrastingly, PBMCs incubated with Zika or attenuated Bonezi et al. Dengue-Mediated Antibody-Secreting Cell DifferentiationYellow Fever viruses showed no correlation between their kynurenine concentrations and ASC numbers. Therefore, our data revealed the existence of distinct pathways for the DENV-mediated ASC differentiation and suggest the involvement of the tryptophan metabolism in this cellular process triggered by flavivirus infections.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Flavivirus-Mediating B Cell Differentiation Into Antibody-Secreting Cells in Humans Is Associated With the Activation of the Tryptophan Metabolism

et al. 2020

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“…Therefore, it was of interest to determine whether the availability of Asn impinged upon T cell metabolism. The tryptophan metabolite kynurenine is transported into T cells via the neutral amino acid transporter Slc7a5/CD98 (21), that also serves as a transporter for essential amino acids tryptophan, leucine, phenylalanine and methionine. We took advantage of the fluorescent properties of kynurenine as well as the glucose analog 2-NB-d-glucose and long chain fatty acid analog BODIPY TM C16 to assess the impact of amino acid deprivation on TCR-induced nutrient uptake.…”

Section: Asn Availability Regulates Tcr-induced Metabolic Reprogrammingmentioning

confidence: 99%

Coordination of asparagine uptake and asparagine synthetase expression is required for T cell activation

Hope

Brownlie

Steele

et al. 2020

Preprint

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T cell receptor triggering by antigen results in metabolic reprogramming that, in turn, facilitates T cells' exit from quiescence. The increased nutrient requirements of activated lymphocytes are met in part by upregulation of cell surface transporters and enhanced uptake of amino acids, fatty acids and glucose from the environment. However, the role of intracellular pathways of amino acid biosynthesis in T cell activation is relatively unexplored. Asparagine (Asn) is a non-essential amino acid that can be synthesized intracellularly through the glutamine-hydrolyzing enzyme asparagine synthetase (ASNS).We set out to define the requirements for uptake of extracellular Asn and ASNS activity in CD8 + T cell activation. At early timepoints of activation, T cells expressed little or no ASNS and, as a consequence, viability and TCR-stimulated growth, activation and metabolic reprogramming were substantially impaired under conditions of Asn deprivation. At later timepoints (>48h of activation), TCR-induced mTOR-dependent signals resulted in upregulation of ASNS, that endowed T cells with the capacity to function independently of extracellular Asn. Thus, we have determined that the coordinated upregulation of ASNS expression and uptake of extracellular Asn is required for optimal T cell effector responses.

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“…Accordingly, we examined the impact of Myc expression on the functional capacity of T cells to transport amino acids and we assessed whether the loss of amino acid transporter expression could recapitulate the striking impact of Myc deletion on T cell protein production. We focused on the system L transporter Slc7a5, as this is the most abundant amino acid transporter expressed on immune activated T lymphocytes ( Fig 3A, 4B, Table S1) and mediates transport of many essential amino acids including methionine, leucine, phenylalanine and tryptophan (Sinclair et al, 2019;Sinclair, Neyens, Ramsay, Taylor, & Cantrell, 2018;Sinclair et al, 2013).…”

Section: Myc Controls Amino Acid Transporter Expression In Immune Actmentioning

confidence: 99%

“…Kynurenine uptake assay were performed as described in (Sinclair et al, 2018). Briefly, antibodies against surface markers were added to culture (37 o C 5% CO2) for 10 minutes prior to uptake assay in order to identify cell types.…”

Section: Kynurenine Uptakesmentioning

confidence: 99%

Quantitative analysis of how Myc controls T cell proteomes and metabolic pathways during T cell activation

Marchingo

Sinclair

Howden

et al. 2019

Preprint

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T cell expansion and differentiation are critically dependent on the transcription factor c-Myc (Myc). Herein we use quantitative mass-spectrometry to reveal how Myc controls antigen receptor driven cell growth and proteome restructuring in T cells. Analysis of copy numbers per cell of >7000 proteins provides new understanding of the selective role of Myc in controlling the protein machinery that govern T cell fate. The data identify both Myc dependent and independent metabolic processes in immune activated T cells. We uncover that a primary function of Myc is to control expression of multiple amino acid transporters and that loss of a single Myc-controlled amino acid transporter effectively phenocopies the impact of Myc deletion. This study provides a comprehensive map of how Myc selectively shapes T cell phenotypes, revealing that Myc induction of amino acid transport is pivotal for subsequent bioenergetic and biosynthetic programs and licences T cell receptor driven proteome reprogramming.

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Single cell analysis of kynurenine and System L amino acid transport in T cells

Cited by 136 publications

References 36 publications

Flavivirus-Mediating B Cell Differentiation Into Antibody-Secreting Cells in Humans Is Associated With the Activation of the Tryptophan Metabolism

Flavivirus-Mediating B Cell Differentiation Into Antibody-Secreting Cells in Humans Is Associated With the Activation of the Tryptophan Metabolism

Coordination of asparagine uptake and asparagine synthetase expression is required for T cell activation

Quantitative analysis of how Myc controls T cell proteomes and metabolic pathways during T cell activation

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