Metabolic but not transcriptional regulation by PKM2 is important for Natural Killer cell responses

  1. Jessica F Walls
  2. Jeff J Subleski
  3. Erika M Palmieri
  4. Marieli Gonzalez Cotto
  5. Clair M Gardiner
  6. Daniel W McVicar
  7. David K Finlay  Is a corresponding author
  1. National Cancer Institute, United States
  2. Trinity College Dublin, Ireland

Abstract

Natural Killer (NK) cells have an important role in immune responses to viruses and tumours. Integrating changes in signal transduction pathways and cellular metabolism is essential for effective NK cells responses. The glycolytic enzyme Pyruvate Kinase Muscle 2 (PKM2) has described roles in regulating glycolytic flux and signal transduction, particularly gene transcription. While PKM2 expression is robustly induced in activated NK cells, mice lacking PKM2 in NK cells showed no defect in NK cell metabolism, transcription or anti-viral responses to MCMV infection. NK cell metabolism was maintained due to compensatory PKM1 expression in PKM2-null NK cells. To further investigate the role of PKM2 we used TEPP-46, which increases PKM2 catalytic activity while inhibiting any PKM2 signalling functions. NK cells activated with TEPP-46 had reduced effector function due to TEPP-46-induced increases in oxidative stress. Overall, PKM2-regulated glycolytic metabolism and redox status, not transcriptional control, facilitate optimal NK cells responses.

Data availability

RNA sequencing data has been uploaded to GEO (GSE156064).

The following data sets were generated

Article and author information

Author details

  1. Jessica F Walls

    Laboratory of Cancer Immunometabolism, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Jeff J Subleski

    Laboratory of Cancer Immunometabolism, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Erika M Palmieri

    Laboratory of Cancer Immunometabolism, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Marieli Gonzalez Cotto

    Laboratory of Cancer Immunometabolism, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Clair M Gardiner

    School of Biochemistry and Immunology, Trinity College Dublin, Dublin, Ireland
    Competing interests
    The authors declare that no competing interests exist.
  6. Daniel W McVicar

    Laboratory of Cancer Immunometabolism, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. David K Finlay

    School of Biochemistry and Immunology and School of Pharmacy and Pharmaceutical Sciences, Trinity College Dublin, Dublin, Ireland
    For correspondence
    finlayd@tcd.ie
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2716-6679

Funding

National Institutes of Health

  • Daniel W McVicar

Wellcome (106811/Z/15/Z)

  • Jessica F Walls

National Institutes of Health

  • Jessica F Walls

National Institutes of Health

  • Erika M Palmieri

National Institutes of Health

  • Marieli Gonzalez Cotto

National Institutes of Health

  • Jeff J Subleski

Science Foundation Ireland (18/ERCS/6005)

  • David K Finlay

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Tiffany Horng, ShanghaiTech University, China

Ethics

Animal experimentation: Mice utilised in Ireland were maintained in compliance with Irish Department of Health and Children regulations and with the approval of the University of Dublin's ethical review board. Mice utilised in the USA were maintained in accordance with institutional guidelines for animal care and use at NCI Frederick, NIH.

Version history

  1. Received: May 21, 2020
  2. Accepted: August 15, 2020
  3. Accepted Manuscript published: August 19, 2020 (version 1)
  4. Version of Record published: September 2, 2020 (version 2)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 3,194
    views
  • 420
    downloads
  • 18
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Jessica F Walls
  2. Jeff J Subleski
  3. Erika M Palmieri
  4. Marieli Gonzalez Cotto
  5. Clair M Gardiner
  6. Daniel W McVicar
  7. David K Finlay
(2020)
Metabolic but not transcriptional regulation by PKM2 is important for Natural Killer cell responses
eLife 9:e59166.
https://doi.org/10.7554/eLife.59166

Share this article

https://doi.org/10.7554/eLife.59166

Further reading

    1. Cell Biology
    Yuki Date, Yukiko Sasazawa ... Shinji Saiki
    Research Article

    The autophagy-lysosome pathway plays an indispensable role in the protein quality control by degrading abnormal organelles and proteins including a-synuclein (aSyn) associated with the pathogenesis of Parkinson's disease (PD). However, the activation of this pathway is mainly by targeting lysosomal enzymic activity. Here, we focused on the autophagosome-lysosome fusion process around the microtubule-organizing center (MTOC) regulated by lysosomal positioning. Through high-throughput chemical screening, we identified 6 out of 1,200 clinically approved drugs enabling the lysosomes to accumulate around the MTOC with autophagy flux enhancement. We further demonstrated that these compounds induce the lysosomal clustering through a JIP4-TRPML1-dependent mechanism. Among them, the lysosomal-clustering compound albendazole promoted the autophagy-dependent degradation of Triton-X-insoluble, proteasome inhibitor-induced aggregates. In a cellular PD model, albendazole boosted insoluble aSyn degradation. Our results revealed that lysosomal clustering can facilitate the breakdown of protein aggregates, suggesting that lysosome-clustering compounds may offer a promising therapeutic strategy against neurodegenerative diseases characterized by the presence of aggregate-prone proteins.

    1. Cell Biology
    Yuhao Wang, Linhao Ruan ... Rong Li
    Research Article

    Mitochondria are the cellular energy hub and central target of metabolic regulation. Mitochondria also facilitate proteostasis through pathways such as the ‘mitochondria as guardian in cytosol’ (MAGIC) whereby cytosolic misfolded proteins (MPs) are imported into and degraded inside mitochondria. In this study, a genome-wide screen in Saccharomyces cerevisiae uncovered that Snf1, the yeast AMP-activated protein kinase (AMPK), inhibits the import of MPs into mitochondria while promoting mitochondrial biogenesis under glucose starvation. We show that this inhibition requires a downstream transcription factor regulating mitochondrial gene expression and is likely to be conferred through substrate competition and mitochondrial import channel selectivity. We further show that Snf1/AMPK activation protects mitochondrial fitness in yeast and human cells under stress induced by MPs such as those associated with neurodegenerative diseases.