QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations
Summary: System-level analysis of single-cell data is rapidly transforming the field of immunometabolism. Given the competitive demand for nutrients in immune microenvironments, there is a need to understand how and when immune cells access these nutrients. Here, we describe a new approach for singl...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-08-01
|
| Series: | Cell Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124723008392 |
| _version_ | 1827855659004067840 |
|---|---|
| author | Leonard R. Pelgrom Gavin M. Davis Simon O’Shaughnessy Emilie J.M. Wezenberg Sander I. Van Kasteren David K. Finlay Linda V. Sinclair |
| author_facet | Leonard R. Pelgrom Gavin M. Davis Simon O’Shaughnessy Emilie J.M. Wezenberg Sander I. Van Kasteren David K. Finlay Linda V. Sinclair |
| author_sort | Leonard R. Pelgrom |
| collection | DOAJ |
| description | Summary: System-level analysis of single-cell data is rapidly transforming the field of immunometabolism. Given the competitive demand for nutrients in immune microenvironments, there is a need to understand how and when immune cells access these nutrients. Here, we describe a new approach for single-cell analysis of nutrient uptake where we use in-cell biorthogonal labeling of a functionalized amino acid after transport into the cell. In this manner, the bona fide active uptake of glutamine via SLC1A5/ASCT2 could be quantified. We used this assay to interrogate the transport capacity of complex immune subpopulations, both in vitro and in vivo. Taken together, our findings provide an easy sensitive single-cell assay to assess which cells support their function via SLC1A5-mediated uptake. This is a significant addition to the single-cell metabolic toolbox required to decode the metabolic landscape of complex immune microenvironments. |
| first_indexed | 2024-03-12T11:54:15Z |
| format | Article |
| id | doaj.art-f8ed02eb6f6c4fc3b2176333693c7e6f |
| institution | Directory Open Access Journal |
| issn | 2211-1247 |
| language | English |
| last_indexed | 2024-03-12T11:54:15Z |
| publishDate | 2023-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Cell Reports |
| spelling | doaj.art-f8ed02eb6f6c4fc3b2176333693c7e6f2023-08-31T05:01:46ZengElsevierCell Reports2211-12472023-08-01428112828QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populationsLeonard R. Pelgrom0Gavin M. Davis1Simon O’Shaughnessy2Emilie J.M. Wezenberg3Sander I. Van Kasteren4David K. Finlay5Linda V. Sinclair6Leiden Institute of Chemistry and the Institute of Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, the NetherlandsSchool of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160 Pearse Street, D02R590 Dublin, IrelandSchool of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160 Pearse Street, D02R590 Dublin, IrelandLeiden Institute of Chemistry and the Institute of Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, the NetherlandsLeiden Institute of Chemistry and the Institute of Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, the Netherlands; Corresponding authorSchool of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160 Pearse Street, D02R590 Dublin, Ireland; School of Pharmacy and Pharmaceutical Sciences, Trinity Biomedical Sciences Institute, Trinity College Dublin, 152-160 Pearse Street, D02R590 Dublin, Ireland; Corresponding authorSchool of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, UK; Corresponding authorSummary: System-level analysis of single-cell data is rapidly transforming the field of immunometabolism. Given the competitive demand for nutrients in immune microenvironments, there is a need to understand how and when immune cells access these nutrients. Here, we describe a new approach for single-cell analysis of nutrient uptake where we use in-cell biorthogonal labeling of a functionalized amino acid after transport into the cell. In this manner, the bona fide active uptake of glutamine via SLC1A5/ASCT2 could be quantified. We used this assay to interrogate the transport capacity of complex immune subpopulations, both in vitro and in vivo. Taken together, our findings provide an easy sensitive single-cell assay to assess which cells support their function via SLC1A5-mediated uptake. This is a significant addition to the single-cell metabolic toolbox required to decode the metabolic landscape of complex immune microenvironments.http://www.sciencedirect.com/science/article/pii/S2211124723008392CP: MetabolismCP: Immunology |
| spellingShingle | Leonard R. Pelgrom Gavin M. Davis Simon O’Shaughnessy Emilie J.M. Wezenberg Sander I. Van Kasteren David K. Finlay Linda V. Sinclair QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations Cell Reports CP: Metabolism CP: Immunology |
| title | QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations |
| title_full | QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations |
| title_fullStr | QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations |
| title_full_unstemmed | QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations |
| title_short | QUAS-R: An SLC1A5-mediated glutamine uptake assay with single-cell resolution reveals metabolic heterogeneity with immune populations |
| title_sort | quas r an slc1a5 mediated glutamine uptake assay with single cell resolution reveals metabolic heterogeneity with immune populations |
| topic | CP: Metabolism CP: Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2211124723008392 |
| work_keys_str_mv | AT leonardrpelgrom quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT gavinmdavis quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT simonoshaughnessy quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT emiliejmwezenberg quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT sanderivankasteren quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT davidkfinlay quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations AT lindavsinclair quasranslc1a5mediatedglutamineuptakeassaywithsinglecellresolutionrevealsmetabolicheterogeneitywithimmunepopulations |