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Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells

Immune surveillance of cancer cells is facilitated by the Natural Killer Group 2D (NKG2D) receptor expressed by different lymphocyte subsets. It recognizes NKG2D ligands that are rarely expressed on healthy cells, but upregulated by tumorigenesis, presenting a target for immunological clearance. The...

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Autores principales: Møller, Sofie H., Mellergaard, Maiken, Madsen, Mikkel, Bermejo, Amaia V., Jepsen, Stine D., Hansen, Marie H., Høgh, Rikke I., Aldana, Blanca I., Desler, Claus, Rasmussen, Lene Juel, Sustarsic, Elahu G., Gerhart-Hines, Zachary, Daskalaki, Evangelia, Wheelock, Craig E., Hiron, Thomas K., Lin, Da, O’Callaghan, Christopher A., Wandall, Hans H., Andresen, Lars, Skov, Søren
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7431954/
https://www.ncbi.nlm.nih.gov/pubmed/32849657
http://dx.doi.org/10.3389/fimmu.2020.01968
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author Møller, Sofie H.
Mellergaard, Maiken
Madsen, Mikkel
Bermejo, Amaia V.
Jepsen, Stine D.
Hansen, Marie H.
Høgh, Rikke I.
Aldana, Blanca I.
Desler, Claus
Rasmussen, Lene Juel
Sustarsic, Elahu G.
Gerhart-Hines, Zachary
Daskalaki, Evangelia
Wheelock, Craig E.
Hiron, Thomas K.
Lin, Da
O’Callaghan, Christopher A.
Wandall, Hans H.
Andresen, Lars
Skov, Søren
author_facet Møller, Sofie H.
Mellergaard, Maiken
Madsen, Mikkel
Bermejo, Amaia V.
Jepsen, Stine D.
Hansen, Marie H.
Høgh, Rikke I.
Aldana, Blanca I.
Desler, Claus
Rasmussen, Lene Juel
Sustarsic, Elahu G.
Gerhart-Hines, Zachary
Daskalaki, Evangelia
Wheelock, Craig E.
Hiron, Thomas K.
Lin, Da
O’Callaghan, Christopher A.
Wandall, Hans H.
Andresen, Lars
Skov, Søren
author_sort Møller, Sofie H.
collection PubMed
description Immune surveillance of cancer cells is facilitated by the Natural Killer Group 2D (NKG2D) receptor expressed by different lymphocyte subsets. It recognizes NKG2D ligands that are rarely expressed on healthy cells, but upregulated by tumorigenesis, presenting a target for immunological clearance. The molecular mechanisms responsible for NKG2D ligand regulation remain complex. Here we report that cancer cell metabolism supports constitutive surface expression of the NKG2D ligand MHC class I chain-related proteins A (MICA). Knockout of the N-glycosylation gene N-acetylglucosaminyltransferase V (MGAT5) in HEK293 cells induced altered metabolism and continuous high MICA surface expression. MGAT5 knockout cells were used to examine the association of cell metabolism and MICA expression through genetic, pharmacological and metabolic assays. Findings were verified in cancer cell lines. Cells with constitutive high MICA expression showed enhanced spare respiratory capacity and elevated mitochondrial efflux of citrate, determined by extracellular flux analysis and metabolomics. MICA expression was reduced by inhibitors of mitochondrial function, FCCP and etomoxir e.g., and depended on conversion of citrate to acetyl-CoA and oxaloacetate by ATP citrate lyase, which was also observed in several cancer cell types. Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) analysis revealed that upregulated MICA transcription was associated with an open chromatin structure at the MICA transcription start site. We identify mitochondria and cytoplasmic citrate as key regulators of constitutive MICA expression and we propose that metabolic reprogramming of certain cancer cells facilitates MICA expression and NKG2D-mediated immune recognition.
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spelling pubmed-74319542020-08-25 Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells Møller, Sofie H. Mellergaard, Maiken Madsen, Mikkel Bermejo, Amaia V. Jepsen, Stine D. Hansen, Marie H. Høgh, Rikke I. Aldana, Blanca I. Desler, Claus Rasmussen, Lene Juel Sustarsic, Elahu G. Gerhart-Hines, Zachary Daskalaki, Evangelia Wheelock, Craig E. Hiron, Thomas K. Lin, Da O’Callaghan, Christopher A. Wandall, Hans H. Andresen, Lars Skov, Søren Front Immunol Immunology Immune surveillance of cancer cells is facilitated by the Natural Killer Group 2D (NKG2D) receptor expressed by different lymphocyte subsets. It recognizes NKG2D ligands that are rarely expressed on healthy cells, but upregulated by tumorigenesis, presenting a target for immunological clearance. The molecular mechanisms responsible for NKG2D ligand regulation remain complex. Here we report that cancer cell metabolism supports constitutive surface expression of the NKG2D ligand MHC class I chain-related proteins A (MICA). Knockout of the N-glycosylation gene N-acetylglucosaminyltransferase V (MGAT5) in HEK293 cells induced altered metabolism and continuous high MICA surface expression. MGAT5 knockout cells were used to examine the association of cell metabolism and MICA expression through genetic, pharmacological and metabolic assays. Findings were verified in cancer cell lines. Cells with constitutive high MICA expression showed enhanced spare respiratory capacity and elevated mitochondrial efflux of citrate, determined by extracellular flux analysis and metabolomics. MICA expression was reduced by inhibitors of mitochondrial function, FCCP and etomoxir e.g., and depended on conversion of citrate to acetyl-CoA and oxaloacetate by ATP citrate lyase, which was also observed in several cancer cell types. Assay for Transposase-Accessible Chromatin using sequencing (ATAC-seq) analysis revealed that upregulated MICA transcription was associated with an open chromatin structure at the MICA transcription start site. We identify mitochondria and cytoplasmic citrate as key regulators of constitutive MICA expression and we propose that metabolic reprogramming of certain cancer cells facilitates MICA expression and NKG2D-mediated immune recognition. Frontiers Media S.A. 2020-08-11 /pmc/articles/PMC7431954/ /pubmed/32849657 http://dx.doi.org/10.3389/fimmu.2020.01968 Text en Copyright © 2020 Møller, Mellergaard, Madsen, Bermejo, Jepsen, Hansen, Høgh, Aldana, Desler, Rasmussen, Sustarsic, Gerhart-Hines, Daskalaki, Wheelock, Hiron, Lin, O’Callaghan, Wandall, Andresen and Skov. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Møller, Sofie H.
Mellergaard, Maiken
Madsen, Mikkel
Bermejo, Amaia V.
Jepsen, Stine D.
Hansen, Marie H.
Høgh, Rikke I.
Aldana, Blanca I.
Desler, Claus
Rasmussen, Lene Juel
Sustarsic, Elahu G.
Gerhart-Hines, Zachary
Daskalaki, Evangelia
Wheelock, Craig E.
Hiron, Thomas K.
Lin, Da
O’Callaghan, Christopher A.
Wandall, Hans H.
Andresen, Lars
Skov, Søren
Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title_full Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title_fullStr Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title_full_unstemmed Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title_short Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells
title_sort cytoplasmic citrate flux modulates the immune stimulatory nkg2d ligand mica in cancer cells
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7431954/
https://www.ncbi.nlm.nih.gov/pubmed/32849657
http://dx.doi.org/10.3389/fimmu.2020.01968
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