1. Cell Biology
  2. Immunology and Inflammation
Download icon

Wiskott Aldrich syndrome protein regulates non-selective autophagy and mitochondrial homeostasis in human myeloid cells

  1. Elizabeth Rivers
  2. Rajeev Rai
  3. Jonas Lӧtscher
  4. Michael Hollinshead
  5. Gasper Markelj
  6. James Thaventheran
  7. Austen JJ Worth
  8. Alessia Cavazza
  9. Christopher Hess
  10. Mona Bajaj-Elliott
  11. Adrian James Thrasher  Is a corresponding author
  1. UCL Institute of Child Health, United Kingdom
  2. University of Basel, Switzerland
  3. University of Cambridge, United Kingdom
  4. University Medical Centre Ljubljana, Slovenia
  5. Great Ormond Street Hospital for Children NHS Foundation Trust, United Kingdom
Research Article
  • Cited 4
  • Views 947
  • Annotations
Cite this article as: eLife 2020;9:e55547 doi: 10.7554/eLife.55547

Abstract

The actin cytoskeletal regulator Wiskott Aldrich syndrome protein (WASp) has been implicated in maintenance of the autophagy-inflammasome axis in innate murine immune cells. Here, we show that WASp deficiency is associated with impaired rapamycin-induced autophagosome formation and trafficking to lysosomes in primary human monocyte-derived macrophages (MDMs). WASp reconstitution in vitro and in WAS patients following clinical gene therapy restores autophagic flux and is dependent on the actin-related protein complex ARP2/3. Induction of mitochondrial damage with CCCP, as a model of selective autophagy, also reveals a novel ARP2/3-dependent role for WASp in formation of sequestrating actin cages and maintenance of mitochondrial network integrity. Furthermore, mitochondrial respiration is suppressed in WAS patient MDMs and unable to achieve normal maximal activity when stressed, indicating profound intrinsic metabolic dysfunction. Taken together, we provide evidence of new and important roles of human WASp in autophagic processes and immunometabolic regulation, which may mechanistically contribute to the complex WAS immunophenotype.

Data availability

All data associated with this study are present in this manuscript and Supporting Files.

Article and author information

Author details

  1. Elizabeth Rivers

    Infection, Immunity and Inflammation Programme, UCL Institute of Child Health, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Rajeev Rai

    Infection, Immunity and Inflammation Programme, UCL Institute of Child Health, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Jonas Lӧtscher

    Department of Biomedicine, University of Basel, Basel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  4. Michael Hollinshead

    Department of Pathology, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  5. Gasper Markelj

    Department of Allergy, Rheumatology and Clinical Immunology, University Children's Hospital, University Medical Centre Ljubljana, Ljubljana, Slovenia
    Competing interests
    The authors declare that no competing interests exist.
  6. James Thaventheran

    Cambridge Institute for Immunology and Infectious Disease, University of Cambridge, Cambridge, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  7. Austen JJ Worth

    Department of Immunology, Great Ormond Street Hospital for Children NHS Foundation Trust, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6803-7385
  8. Alessia Cavazza

    Infection, Immunity and Inflammation Programme, UCL Institute of Child Health, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  9. Christopher Hess

    Department of Biomedicine, University of Basel, Basel, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  10. Mona Bajaj-Elliott

    Infection, Immunity and Inflammation Programme, UCL Institute of Child Health, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Adrian James Thrasher

    Infection, Immunity and Inflammation Programme, UCL Institute of Child Health, London, United Kingdom
    For correspondence
    a.thrasher@ucl.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6097-6115

Funding

Wellcome Trust (090233/Z/09/Z)

  • Adrian James Thrasher

Wellcome Trust (201250/Z/16/Z)

  • Elizabeth Rivers

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

Ethics

Human subjects: For usage of human CD34+ HSPC from healthy and WAS donors, informed written consent was obtained in accordance with the Declaration of Helsinki and ethical approval from the Great Ormond Street Hospital for Children NHS Foundation Trust and the Institute of Child Health Research Ethics (08/H0713/87).

Reviewing Editor

  1. Tiffany Horng, ShanghaiTech University, China

Publication history

  1. Received: January 28, 2020
  2. Accepted: October 31, 2020
  3. Accepted Manuscript published: November 2, 2020 (version 1)
  4. Version of Record published: November 18, 2020 (version 2)

Copyright

© 2020, Rivers 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

  • 947
    Page views
  • 146
    Downloads
  • 4
    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)

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

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

Further reading

    1. Cell Biology
    2. Structural Biology and Molecular Biophysics
    Carolina Franco Nitta et al.
    Research Article Updated

    Crosstalk between different receptor tyrosine kinases (RTKs) is thought to drive oncogenic signaling and allow therapeutic escape. EGFR and RON are two such RTKs from different subfamilies, which engage in crosstalk through unknown mechanisms. We combined high-resolution imaging with biochemical and mutational studies to ask how EGFR and RON communicate. EGF stimulation promotes EGFR-dependent phosphorylation of RON, but ligand stimulation of RON does not trigger EGFR phosphorylation – arguing that crosstalk is unidirectional. Nanoscale imaging reveals association of EGFR and RON in common plasma membrane microdomains. Two-color single particle tracking captured formation of complexes between RON and EGF-bound EGFR. Our results further show that RON is a substrate for EGFR kinase, and that transactivation of RON requires formation of a signaling competent EGFR dimer. These results support a role for direct EGFR/RON interactions in propagating crosstalk, such that EGF-stimulated EGFR phosphorylates RON to activate RON-directed signaling.

    1. Cell Biology
    2. Neuroscience
    Angela Kim et al.
    Research Article Updated

    Insulin-induced hypoglycemia is a major treatment barrier in type-1 diabetes (T1D). Accordingly, it is important that we understand the mechanisms regulating the circulating levels of glucagon. Varying glucose over the range of concentrations that occur physiologically between the fed and fuel-deprived states (8 to 4 mM) has no significant effect on glucagon secretion in the perfused mouse pancreas or in isolated mouse islets (in vitro), and yet associates with dramatic increases in plasma glucagon. The identity of the systemic factor(s) that elevates circulating glucagon remains unknown. Here, we show that arginine-vasopressin (AVP), secreted from the posterior pituitary, stimulates glucagon secretion. Alpha-cells express high levels of the vasopressin 1b receptor (V1bR) gene (Avpr1b). Activation of AVP neurons in vivo increased circulating copeptin (the C-terminal segment of the AVP precursor peptide) and increased blood glucose; effects blocked by pharmacological antagonism of either the glucagon receptor or V1bR. AVP also mediates the stimulatory effects of hypoglycemia produced by exogenous insulin and 2-deoxy-D-glucose on glucagon secretion. We show that the A1/C1 neurons of the medulla oblongata drive AVP neuron activation in response to insulin-induced hypoglycemia. AVP injection increased cytoplasmic Ca2+ in alpha-cells (implanted into the anterior chamber of the eye) and glucagon release. Hypoglycemia also increases circulating levels of AVP/copeptin in humans and this hormone stimulates glucagon secretion from human islets. In patients with T1D, hypoglycemia failed to increase both copeptin and glucagon. These findings suggest that AVP is a physiological systemic regulator of glucagon secretion and that this mechanism becomes impaired in T1D.