Limitations of PLX3397 as a microglial investigational tool: peripheral and off-target effects dictate the response to inflammation

Front Immunol. 2023 Nov 22:14:1283711. doi: 10.3389/fimmu.2023.1283711. eCollection 2023.

Abstract

Microglia, the resident macrophages of the central nervous system (CNS), play a critical role in CNS homeostasis and neuroinflammation. Pexidartinib (PLX3397), a colony-stimulating factor 1 (CSF1) receptor inhibitor, is widely used to deplete microglia, offering flexible options for both long-term depletion and highly versatile depletion-repopulation cycles. However, the potential impact of PLX3397 on peripheral (immune) cells remains controversial. Until now, the microglia-specificity of this type of compounds has not been thoroughly evaluated, particularly in the context of peripherally derived neuroinflammation. Our study addresses this gap by examining the effects of PLX3397 on immune cells in the brain, liver, circulation and bone marrow, both in homeostasis and systemic inflammation models. Intriguingly, we demonstrate that PLX3397 treatment not only influences the levels of tissue-resident macrophages, but also affects circulating and bone marrow immune cells beyond the mononuclear phagocyte system (MPS). These alterations in peripheral immune cells disrupt the response to systemic inflammation, consequently impacting the phenotype irrespective of microglial depletion. Furthermore, we observed that a lower dose of PLX3397, which does not deplete microglia, demonstrates similar (non-)MPS effects, both in the periphery and the brain, but fails to fully replicate the peripheral alterations seen in the higher doses, questioning lower doses as a 'peripheral control' strategy. Overall, our data highlight the need for caution when interpreting studies employing this compound, as it may not be suitable for specific investigation of microglial function in the presence of systemic inflammation.

Keywords: PLX3397; bile duct ligation (BDL); endotoxemia; lipopolysaccharide (LPS); microglia; neuroinflammation; peripheral inflammation.

MeSH terms

  • Brain
  • Humans
  • Inflammation / drug therapy
  • Microglia*
  • Neuroinflammatory Diseases*

Substances

  • pexidartinib

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. WC, LH, DV and FP are supported by a grants from the Research Foundation – Flanders (11A6420N, 1268823N, 11L2122N, 11D0522N). CS is a Francqui Foundation Professor and supported by an ERC starting grant (MyeFattyLiver, 851908). AG is a senior clinical researcher of the Research Foundation – Flanders (1805718N). RV is supported by a FWO junior research project grant (G055121N). GI is supported by a grant from the Stichting voor Alzheimer onderzoek (SAO) (20200032). This project is supported by VIB.