The Alzheimer’s disease protective P522R variant of PLCG2 , consistently enhances stimulus-dependent PLCγ2 activation, depleting substrate and altering cell function
Open accessAbstract
Recent genome-wide association studies of Alzheimer’s disease (AD) have identified variants implicating immune pathways in disease development. A rare coding variant of PLCG2 , which encodes PLCγ2, shows a significant protective effect for AD (rs72824905, P522R, P =5.38×10 −10 , Odds Ratio = 0.68). Molecular dynamic modelling of the PLCγ2-R522 variant, situated within the auto-inhibitory domain of PLCγ2, suggests a structural change to the protein. Through CRISPR-engineering we have generated novel PLCG2 -R522 harbouring human induced pluripotent cell lines (hiPSC) and a mouse knockin model, neither of which exhibits alterations in endogenous PLCG2 expression. Mouse microglia and macrophages and hiPSC-derived microglia-like cells with the R522 mutation, all demonstrate a consistent non-redundant hyperfunctionality in the context of normal expression of other PLC isoforms. This signalling alteration manifests as enhanced cellular Ca 2+ store release (∼20-40% increase) in response to physiologically-relevant stimuli (e.g. Fc receptor ligation and Aβ oligomers). This hyperfunctionality resulted in increased PIP 2 depletion in the cells with the PLCγ2-R522 variant after exposure to stimuli and reduced basal detection of PIP 2 levels in vivo . These PLCγ2-R522 associated abnormalities resulted in impairments to phagocytosis (fungal and bacterial particles) and enhanced endocytosis (Aβ oligomers and dextran). PLCγ2 sits downstream of disease relevant pathways, such as TREM2 and CSF1R and alterations in its activity, direct impacts cell function, which in the context of the inherent drugability of enzymes such as PLCγ2, raise the prospect of manipulation of PLCγ2 as a therapeutic target in Alzheimer’s Disease.
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PLCγ2 p.P522R is associated with reduction of cholesterol ester accumulation and increased phagocytosis.
“reduction of cholesterol ester accumulation,Increased phagocytosis [36]”
From TREM2/PLCγ2 signalling in immune cells: function, structural insight, and potential therapeutic modulation (2021) · cites this paper as [36]
The Results section, “Expression of the AD protective PLCγ2 R522 variant impaired phagocytic and enhanced endocytic clearance” (page unavailable), reports significantly reduced—not increased—phagocytosis in R522-expressing cells compared with P522 controls. The paper reports reduced PI(4,5)P2 levels, but the passages found do not establish reduced cholesterol ester accumulation. (LLM, conf=0.90)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredThe PLCγ2 p.P522R variant causes an increase in microglial phagocytosis and survival, whereas TREM2 LOAD variants typically induce a reduction in these phenotypes.
“Whilst TREM2 LOAD variants typically induce a reduction in microglial phagocytosis and survival, the PLCγ2 p.P522R variant causes an increase in these beneficial microglial phenotypes [35, 36]”
From TREM2/PLCγ2 signalling in immune cells: function, structural insight, and potential therapeutic modulation (2021) · cites this paper as [36]
The Results section reports that microglia expressing PLCγ2 R522 had significantly reduced, not increased, phagocytosis of E. coli and zymosan particles compared with P522 controls. The paper reports enhanced endocytic clearance of Aβ oligomers, but that is a different phenotype; the cited passages do not establish increased microglial survival or the claimed effects of TREM2 LOAD variants. (LLM, conf=0.90)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredIn human iPSC-derived macrophages, intracellular calcium was elevated following PLCγ2 stimulation with an FcγRII-activating antibody in P522R variant lines compared with WT.
“data in human iPSC-derived macrophages has corroborated this finding, demonstrating that intracellular calcium was elevated following PLCγ2 stimulation with an FcγRII-activating antibody in the P522R variant lines, when compared with the WT [36]”
From TREM2/PLCγ2 signalling in immune cells: function, structural insight, and potential therapeutic modulation (2021) · cites this paper as [36]
The Results subsection on receptor-mediated Ca²⁺ release (Fig. 2C,D; page not provided) reports a greater cytosolic Ca²⁺ increase after anti-CD32 (anti-FcγRII) stimulation in human iPSC-derived R522 cells than in P522 controls. It identifies those human cells as microglia-like cells, not macrophages; the macrophage calcium experiments used mouse-derived cells. (LLM, conf=0.88)
Experimental — accuracy not yet measuredp.P522R increased Aβ-oligomer clearance compared with WT expressing microglia, macrophages, and human derived iPSC macrophages.
“Maguire et al. demonstrated that p.P522R increased Aβ-oligomer clearance when compared with WT expressing microglia, macrophages, and human derived iPSC macrophages [36]”
From TREM2/PLCγ2 signalling in immune cells: function, structural insight, and potential therapeutic modulation (2021) · cites this paper as [36]
The clearance-results section reports enhanced Aβ1-42 oligomer clearance with R522 in mouse macrophages, mouse microglia, and human iPSC-derived microglia. The claim calls the human iPSC-derived cells macrophages, which is not the cell type reported in the paper. (LLM, conf=0.89)
Experimental — accuracy not yet measuredIn mouse microglia, basal levels of PIP2 may be decreased in cells harbouring p.P522R, suggesting that in mouse brain the P522R mutation might elicit an increase in basal PLCγ2 function.
“Recent studies in mouse microglia have indicated that basal levels of PIP2 may be decreased in cells harbouring the p.P522R variant, suggesting that in the mouse brain the P522R mutation might elicit an increase in basal PLCγ2 function [36]”
From TREM2/PLCγ2 signalling in immune cells: function, structural insight, and potential therapeutic modulation (2021) · cites this paper as [36]
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Magno, L., Bunney, T. D., Mead, E., Svensson, F., & Bictash, M. N. (2021) · Molecular Neurodegeneration · doi:10.1186/s13024-021-00436-5 · Cited by 64 · see citing works
Obst, J., Hall-Roberts, H. L., Smith, T. B., Kreuzer, M., Magno, L., Di Daniel, E., Davis, J. B., & Mead, E. (2021) · Scientific Reports · doi:10.1038/s41598-021-96144-7 · Cited by 36 · see citing works
- Authors
- Maguire, E., Menzies, G. E., Phillips, T., Sasner, M., Williams, H. M., Czubala, M. A., Evans, N., Cope, E. L., Sims, R., Howell, G. R., Lloyd-Evans, E., Williams, J., Allen, N. D., & Taylor, P. R.
- Affiliations
- UK Dementia Research Institute (GB)
- Jackson Laboratory (US)
- Cardiff University (GB)
- Corresponding author
- Taylor, Philip R.
- ORCID
- Venue
- bioRxiv (Cold Spring Harbor Laboratory)
- Publisher
- openRxiv
- Published
- 2020-04-28
- OpenAlex
- W3023718274
- Cited by
- 4
- References
- 10
- Citations by year
- 2025: 1 · 2021: 2 · 2020: 1
- Type
- posted content
- Language
- en
- Open access
- green
- doi.org
- Licence
- Locations
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- Funders
- Wellcome Trust (107964, 107964/Z/15/Z, /Z/15/Z)
- Llywodraeth Cymru
- Eisai
- UK Dementia Research Institute
- Medical Research Council (MR/N013255/1, MR/L501517/1, MR/K013041/1, MC_PC_17112, N013255/1)
- Biotechnology and Biological Sciences Research Council (BB/T009543/1)
- European Regional Development Fund
- Topics
- Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
- Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- Alzheimer's disease, PLCγ2, autoinhibitory domain, calcium store release, microglia, macrophages, phagocytosis, endocytosis, amyloid-beta oligomers, TREM2, CSF1R, induced pluripotent stem cells, protective genetic variants, therapeutic target
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