Microglia determine an immune-challenged environment and facilitate ibuprofen action in human retinal organoids.

Abstract

Prenatal immune challenges pose significant risks to human embryonic brain and eye development. However, we still lack knowledge about the safe usage of anti-inflammatory drugs during pregnancy. While human induced pluripotent stem cells (hIPSC)-derived brain organoid models have started to explore functional consequences upon viral stimulation, these models commonly lack microglia. Yet, microglia are susceptible to and promote inflammation and will influence the effects. Here, we generate hIPSC-derived microglia precursor cells and assemble them into retinal organoids to dissect their interplay with the developing nervous system and expose them to the immunostimulant poly(I:C). We first identified successful hIPSC-derived microglia (iMG) integration into the retinal organoid at the time when the outer plexiform layer forms. To improve ganglion cell surveillance, we adapted the retinal organoid model and found that the ganglion cell number significantly decreased only with iMG presence. While poly(I:C) exposure alters the iMG phenotype, it does not hinder their interaction with ganglion cells. Furthermore, iMG significantly enhances the supernatants inflammatory secretome and increases retinal cell proliferation. Simultaneous exposure with the non-steroidal anti-inflammatory drug (NSAID) ibuprofen dampens the poly(I:C)-mediated changes of the iMG phenotype and ameliorates cell proliferation. Remarkably, while poly(I:C) disrupts neuronal calcium dynamics independent of iMG, ibuprofen rescues this effect only if iMG are present. This effect depends on cyclooxygenase 1 in microglia and cyclooxygenase 2, common NSAID targets. These findings underscore the importance of microglia in the context of prenatal immune challenges and provide insight into the mechanisms by which ibuprofen exerts its protective effects during embryonic development.