Microglial GRB2 is essential for brain ventriculogenesis and cerebrospinal fluid homeostasis

Microglia play essential yet poorly understood roles in brain development, including axon guidance, regulation of neurogenesis, and pruning of neuronal projections. Congenital hydrocephalus (CH), characterized by enlarged ventricles filled with cerebrospinal fluid (CSF), is a leading cause of pediatric brain surgery, but its molecular mechanisms remain unclear. We have identified what we believe to be novel, recurrent, damaging missense variants in the SH3-binding domain of the adaptor protein growth factor receptor-bound protein 2 (GRB2) in unrelated patients with CH. GRB2 is significantly coexpressed with 1 of its known upstream RTK partners, CSF1R, in the developing human brain, particularly in a microglial subtype associated with regulation of neural stem cells. IP validated GRB2-CSF1R binding in mouse microglial cells and a human monocyte cell line. Cx3cr1-Grb2fl/fl mice engineered with conditional deletion of Grb2 in microglia exhibit congenital absence of microglia and early postnatal severe communicating (nonobstructive) hydrocephalus, mimicking the condition of patients with the GRB2 mutation. The severe ventriculomegaly of Cx3cr1-Grb2fl/fl mice is associated with both depletion of cerebral cortical neurons and impairment of glia-lymphatic-mediated CSF flow. Together, these findings implicate a role of GRB2 in microglia that could be essential for brain development and CSF homeostasis.