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PLCγ2 regulates TREM2 signalling and integrin-mediated adhesion and migration of human iPSC-derived macrophages

Juliane Obst1, Hazel L Hall-Roberts2,3,4, Thomas B Smith2

  • 1Alzheimer's Research UK Oxford Drug Discovery Institute, Centre for Medicines Discovery, University of Oxford, Oxford, UK. juliane.obst@cmd.ox.ac.uk.

Scientific Reports
|October 7, 2021
PubMed

Insights

Phospholipase C gamma 2 (PLCγ2) is crucial for TREM2 signaling in brain immune cells, impacting Alzheimer's disease (AD) pathology. Its absence impairs microglial functions like calcium signaling and phagocytosis, suggesting PLCγ2 as a therapeutic target for AD.

Area of Science:

  • Neuroimmunology
  • Cellular Biology
  • Biochemistry

Background:

  • Genetic studies link microglial genes, including TREM2 and PLCG2, to Alzheimer's disease (AD) risk.
  • A specific PLCG2 variant (P522R) is associated with AD protection and increased enzymatic activity.
  • PLCγ2 mediates intracellular calcium (Ca2+) signals downstream of myeloid cell receptors like TREM2.

Purpose of the Study:

  • To investigate the functional relationship between PLCγ2 and TREM2 signaling.
  • To elucidate the role of PLCγ2 in regulating macrophage and microglial immune functions.

Main Methods:

  • Generated isogenic human induced pluripotent stem cell (iPSC)-derived macrophages with homozygous PLCG2 knockout (Ko).
  • Stimulated TREM2 signaling using a polyclonal antibody to assess calcium flux and inositol trisphosphate (IP1) accumulation.
  • Analyzed macrophage surface marker expression, phagocytic activity, survival, cytokine secretion (TNFα, IL-6), adhesion, and migration.

Main Results:

  • PLCγ2 knockout cells showed a complete absence of calcium flux and IP1 accumulation downstream of TREM2 stimulation, indicating a non-redundant role.
  • Loss of PLCγ2 resulted in altered macrophage surface markers, reduced phagocytosis, and decreased cell survival.
  • Deficiencies in cellular adhesion and migration were observed in PLCγ2-deficient cells.
  • Lipopolysaccharide (LPS)-induced secretion of TNFα and IL-6 remained unaffected.

Conclusions:

  • PLCγ2 is essential for TREM2-mediated calcium signaling and critical for diverse microglial functions, including phagocytosis, survival, adhesion, and migration.
  • Targeting PLCγ2 may offer a strategy to enhance beneficial microglial functions in Alzheimer's disease.

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