Functional Genome-wide Screen Identifies Pathways Restricting Central Nervous System Axonal Regeneration

Abstract

Summary: Axonal regrowth is crucial for recovery from CNS injury but is severely restricted in adult mammals. We used a genome-wide loss-of-function screen for factors limiting axonal regeneration from cerebral cortical neurons in vitro. Knockdown of 16,007 individual genes identified 580 significant phenotypes. These molecules share no significant overlap with those suggested by previous expression profiles. There is enrichment for genes in pathways related to transport, receptor binding, and cytokine signaling, including Socs4 and Ship2. Among transport-regulating proteins, Rab GTPases are prominent. In vivo assessment with C. elegans validates a cell-autonomous restriction of regeneration by Rab27. Mice lacking Rab27b show enhanced retinal ganglion cell axon regeneration after optic nerve crush and greater motor function and raphespinal sprouting after spinal cord trauma. Thus, a comprehensive functional screen reveals multiple pathways restricting axonal regeneration and neurological recovery after injury. : Sekine et al. conduct a genome-wide loss-of-function screen for factors limiting the success of CNS axonal regeneration in mice. They uncover a role for transport, receptor binding, and cytokine signaling pathways. In particular, in vivo loss of Rab27b expression increases axonal regeneration in worm and mouse optic nerve. Keywords: axon, spinal cord injury, CNS trauma, axonal regeneration, rab27, shRNA screen, optic nerve regeneration