Discovery of surrogate agonists for visceral fat Treg cells that modulate metabolic indices in vivo

  1. Ricardo A Fernandes
  2. Chaoran Li
  3. Gang Wang
  4. Xinbo Yang
  5. Christina S Savvides
  6. Caleb R Glassman
  7. Shen Dong
  8. Eric Luxenberg
  9. Leah V Sibener
  10. Michael E Birnbaum
  11. Christophe Benoist
  12. Diane Mathis  Is a corresponding author
  13. K Christopher Garcia  Is a corresponding author
  1. Stanford University School of Medicine, United States
  2. Harvard Medical School, United States
  3. Stanford University School of Engineering, United States
  4. Howard Hughes Medical Institute, Stanford University School of Medicine, United States

Abstract

T regulatory (Treg) cells play vital roles in modulating immunity and tissue homeostasis. Their actions depend on TCR recognition of peptide-MHC molecules; yet the degree of peptide specificity of Treg-cell function, and whether Treg ligands can be used to manipulate Treg cell biology are unknown. Here, we developed an Ab-peptide library that enabled unbiased screening of peptides recognized by a bona fide murine Treg cell clone isolated from the visceral adipose tissue (VAT), and identified surrogate agonist peptides, with differing affinities and signaling potencies. The VAT-Treg cells expanded in vivo by one of the surrogate agonists preserved the typical VAT-Treg transcriptional programs. Immunization with this surrogate, especially when coupled with blockade of TNFa signaling, expanded VAT-Treg cells, resulting in protection from inflammation and improved metabolic indices, including promotion of insulin sensitivity. These studies suggest that antigen-specific targeting of VAT-localized Treg cells could eventually be a strategy for improving metabolic disease.

Data availability

Sequencing data for the peptide-Ab yeast library screening and RNA-seq data for VAT-Treg cells have been deposited in GEO under accession codes GSE151070 and GSE150173. Custom Perl scripts for the processing of the deep sequencing data for the peptide-Ab is available from: https://github.com/jlmendozabio/NGSpeptideprepandpred.

The following data sets were generated

Article and author information

Author details

  1. Ricardo A Fernandes

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Chaoran Li

    Department of Immunology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Gang Wang

    Department of Immunology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Xinbo Yang

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Christina S Savvides

    Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Caleb R Glassman

    Molecular & Cellular Physiology and Structural Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Shen Dong

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Eric Luxenberg

    Department of Electrical Engineering, Stanford University School of Engineering, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Leah V Sibener

    Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Michael E Birnbaum

    Molecular & Cellular Physiology and Structural Biology, Stanford University School of Medicine, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Christophe Benoist

    Department of Immunology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Diane Mathis

    Department of Immunology, Harvard Medical School, Boston, United States
    For correspondence
    diane_mathis@hms.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
  13. K Christopher Garcia

    Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, United States
    For correspondence
    kcgarcia@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9273-0278

Funding

Wellcome (WT101609MA)

  • Ricardo A Fernandes

NIH Office of the Director (5R01AI103867)

  • K Christopher Garcia

Howard Hughes Medical Institute (HHMI)

  • K Christopher Garcia

G Harold and Leila Y. Mathers Foundation

  • K Christopher Garcia

NIH Clinical Center (2R01 DK092541)

  • Diane Mathis

JPB Foundation

  • Diane Mathis

NIH Office of the Director (UC4DK116264)

  • K Christopher Garcia

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

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and every effort was made to minimize suffering. All experiments were performed following animal protocols approved by the HMS Institutional Animal Use and Care Committee (protocol IS00001257).

Reviewing Editor

  1. Bernard Malissen, Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, France

Publication history

  1. Received: April 30, 2020
  2. Accepted: August 4, 2020
  3. Accepted Manuscript published: August 10, 2020 (version 1)
  4. Accepted Manuscript updated: August 17, 2020 (version 2)
  5. Version of Record published: August 20, 2020 (version 3)

Copyright

© 2020, Fernandes et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,927
    Page views
  • 351
    Downloads
  • 10
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

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. Ricardo A Fernandes
  2. Chaoran Li
  3. Gang Wang
  4. Xinbo Yang
  5. Christina S Savvides
  6. Caleb R Glassman
  7. Shen Dong
  8. Eric Luxenberg
  9. Leah V Sibener
  10. Michael E Birnbaum
  11. Christophe Benoist
  12. Diane Mathis
  13. K Christopher Garcia
(2020)
Discovery of surrogate agonists for visceral fat Treg cells that modulate metabolic indices in vivo
eLife 9:e58463.
https://doi.org/10.7554/eLife.58463
  1. Further reading

Further reading

    1. Computational and Systems Biology
    2. Immunology and Inflammation
    Harry Kane, Nelson M LaMarche ... Lydia Lynch
    Research Article

    Innate T cells, including CD1d-restricted invariant natural killer T (iNKT) cells, are characterized by their rapid activation in response to non-peptide antigens, such as lipids. While the transcriptional profiles of naive, effector and memory adaptive T cells have been well studied, less is known about transcriptional regulation of different iNKT cell activation states. Here, using single cell RNA-sequencing, we performed longitudinal profiling of activated murine iNKT cells, generating a transcriptomic atlas of iNKT cell activation states. We found that transcriptional signatures of activation are highly conserved among heterogeneous iNKT cell populations, including NKT1, NKT2 and NKT17 subsets, and human iNKT cells. Strikingly, we found that regulatory iNKT cells, such as adipose iNKT cells, undergo blunted activation, and display constitutive enrichment of memory-like cMAF+ and KLRG1+ populations. Moreover, we identify a conserved cMAF-associated transcriptional network among NKT10 cells, providing novel insights into the biology of regulatory and antigen experienced iNKT cells.

    1. Cancer Biology
    2. Immunology and Inflammation
    Lei Yang, Xichen Dong ... Zhenjun Wang
    Research Article

    Efficacy of immunotherapy is limited in patients with colorectal cancer (CRC) because high expression of tumor-derived transforming growth factor (TGF)-β pathway molecules and interferon (IFN)-stimulated genes (ISGs) promotes tumor immune evasion. Here, we identified a long noncoding RNA (lncRNA), VPS9D1-AS1, which was located in ribosomes and amplified TGF-β signaling and ISG expression. We show that high expression of VPS9D1-AS1 was negatively associated with T lymphocyte infiltration in two independent cohorts of CRC. VPS9D1-AS1 served as a scaffolding lncRNA by binding with ribosome protein S3 (RPS3) to increase the translation of TGF-β, TGFBR1, and SMAD1/5/9. VPS9D1-AS1 knockout downregulated OAS1, an ISG gene, which further reduced IFNAR1 levels in tumor cells. Conversely, tumor cells overexpressing VPS9D1-AS1 were resistant to CD8+ T cell killing and lowered IFNAR1 expression in CD8+ T cells. In a conditional overexpression mouse model, VPS9D1-AS1 enhanced tumorigenesis and suppressed the infiltration of CD8+ T cells. Treating tumor-bearing mice with antisense oligonucleotide drugs targeting VPS9D1-AS1 significantly suppressed tumor growth. Our findings indicate that the tumor-derived VPS9D1-AS1/TGF-β/ISG signaling cascade promotes tumor growth and enhances immune evasion and may thus serve as a potential therapeutic target for CRC.