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Cited by 62Neuroinflammation and Neurodegeneration MechanismsAlzheimer's disease research and treatmentsInflammasome and immune disorders

The DOI matches its Crossref record; 34 of the 34 references it cites were found in a registry or an index.

TREM2 Alzheimer’s variant R47H causes similar transcriptional dysregulation to knockout, yet only subtle functional phenotypes in human iPSC-derived macrophages

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Hall-Roberts, Hazel, Agarwal, Devika, Obst, Juliane, Smith, Thomas B., Monzón-Sandoval, Jimena, Di Daniel, Elena, Webber, Caleb, James, William S., Mead, Emma, Davis, John B., Cowley, Sally A.·2020·Alzheimer's Research & Therapy
DOI 10.1186/s13195-020-00709-zPMC PMC7667762

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Abstract

Background: TREM2 is a microglial cell surface receptor, with risk mutations linked to Alzheimer’s disease (AD), including R47H. TREM2 signalling via SYK aids phagocytosis, chemotaxis, survival, and changes to microglial activation state. In AD mouse models, knockout (KO) of TREM2 impairs microglial clustering around amyloid and prevents microglial activation. The R47H mutation is proposed to reduce TREM2 ligand binding. We investigated cell phenotypes of the R47H mutant and TREM2 KO in a model of human microglia, and compared their transcriptional signatures, to determine the mechanism by which R47H TREM2 disrupts function. Methods: We generated human microglia-like iPSC-macrophages (pMac) from isogenic induced pluripotent stem cell (iPSC) lines, with homozygous R47H mutation or TREM2 knockout (KO). We firstly validated the effect of the R47H mutant on TREM2 surface and subcellular localization in pMac. To assess microglial phenotypic function, we measured phagocytosis of dead neurons, cell morphology, directed migration, survival, and LPS-induced inflammation. We performed bulk RNA-seq, comparing significant differentially expressed genes (DEGs; p < 0.05) between the R47H and KO versus WT, and bioinformatically predicted potential upstream regulators of TREM2-mediated gene expression. Results: R47H modified surface expression and shedding of TREM2, but did not impair TREM2-mediated signalling, or gross phenotypes that were dysregulated in the TREM2 KO (phagocytosis, motility, survival). However, altered gene expression in the R47H TREM2 pMac overlapped by 90% with the TREM2 KO and was characterised by dysregulation of genes involved with immunity, proliferation, activation, chemotaxis, and adhesion. Downregulated mediators of ECM adhesion included the vitronectin receptor αVβ3, and consequently, R47H TREM2 pMac adhered weakly to vitronectin compared with WT pMac. To counteract these transcriptional defects, we investigated TGFβ1, as a candidate upstream regulator. TGFβ1 failed to rescue vitronectin adhesion of pMac, although it improved αVβ3 expression. Conclusions: The R47H mutation is not sufficient to cause gross phenotypic defects of human pMac under standard culture conditions. However, overlapping transcriptional defects with TREM2 KO supports the hypothesised partial loss-of-function effects of the R47H mutation. Furthermore, transcriptomics can guide us to more subtle phenotypic defects in the R47H cells, such as reduced cell adhesion, and can be used to predict targets for therapeutic intervention.

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Crossref · 10.1186/s13195-020-00709-z

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  • A TREM2-dependent survival defect has been shown in TREM2 Ko iPSC-derived microglia-like cells, a deficit also present in cells lacking PLCγ2.

    “Previously, a TREM2-dependent survival defect has been shown in TREM2 Ko iPSC-derived microglia-like cells23,26, a deficit that is also present in cells lacking PLCγ226.”

    From PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages (2021) · cites this paper as [23]

    The Results section (page not available) reports a small but significant increase in cell death in TREM2-knockout iPSC-derived macrophages after M-CSF withdrawal, supporting a survival defect in this microglia-like model under that condition. The paper does not report a survival defect in PLCγ2-deficient cells; its Discussion mentions PLCG2 knockout in connection with shared dysregulated genes, not survival. (LLM, conf=0.88)

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  • The TREM2 activating antibody AF1828 induces downstream signalling including SYK activation and release of calcium from intracellular stores.

    “In order to stimulate TREM2 we used a TREM2 activating antibody (R&D systems, AF1828) that induces downstream signalling including SYK activation and release of calcium from intracellular stores23,31.”

    From PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages (2021) · cites this paper as [23]

  • TREM2 is a cell-surface receptor implicated in microglial functions such as phagocytosis, chemotaxis and survival.

    “TREM2 is a cell-surface receptor of the immunoglobulin superfamily implicated in a variety of microglial functions, such as phagocytosis, chemotaxis and survival21–23.”

    From PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages (2021) · cites this paper as [23]

  • Both synaptosomes and dead SHSY5Y expose surface phosphatidylserine and are taken up by iPSC-derived macrophages at least partially via TREM2-dependent mechanisms.

    “Both substrates expose surface phosphatidylserine and were recently shown to be taken up by iPSC-derived macrophages at least partially via TREM2-dependent mechanisms23.”

    From PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages (2021) · cites this paper as [23]

  • Genetic deficiency of PLCγ2 significantly reduced phagocytosis of cortical synaptosomes and dead cells.

    “Similarly to TREM2 Ko macrophages23, genetic deficiency of PLCγ2 significantly reduced phagocytosis of both cargos”

    From PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages (2021) · cites this paper as [23]

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Metadata

Authors
Hall-Roberts, H., Agarwal, D., Obst, J., Smith, T. B., Monzón-Sandoval, J., Di Daniel, E., Webber, C., James, W. S., Mead, E., Davis, J. B., & Cowley, S. A.
Affiliations
  • University of Oxford (GB)
  • UK Dementia Research Institute (GB)
Corresponding author
Cowley, Sally A.
ORCID
Venue
Alzheimer's Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2020-11-16
Volume
12
Issue
1
Pages
151-151
ISSN
1758-9193 (ISSN-L)
DOI
10.1186/s13195-020-00709-z
PMC
PMC7667762
OpenAlex
W3100678887
Cited by
62
References
34
Citations by year
2026: 8 · 2025: 10 · 2024: 13 · 2023: 8 · 2022: 13 · 2021: 8 · 2020: 2
FWCI
3.00
Type
journal article
Language
en
Open access
Licence
Locations
  • Alzheimer s Research & Therapy (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Alzheimer’s Research UK Oxford Drug Discovery Institute (ARUK-2020DDI-OX)
  • Oxford Martin School, University of Oxford (LC0910-004)
  • Parkinson’s UK (J-1403)
  • Medical Research Council (MC_EX_MR/N50192X/1, MR/N013255/1, MC_PC_16034, G0900747 91070)
  • Wellcome Trust (090532/Z/09/Z, 203141/Z/16/Z)
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Alzheimer's disease research and treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
Keywords
TREM2, Alzheimer's disease, microglia, iPSC-derived macrophages, transcriptional dysregulation, TGF-β1, microglial activation, phagocytosis, lipopolysaccharide-induced inflammation, gene regulatory networks
MeSH terms
Alzheimer Disease, Brain, Humans, Macrophages, Membrane Glycoproteins, Phenotype, Receptors, Immunologic, Microglia, Induced Pluripotent Stem Cells

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