Adaptor protein (AP) complexes have critical roles in transmembrane protein sorting. AP-4 remains poorly understood in the brain despite its loss of function leading to a hereditary spastic paraplegia termed AP-4 deficiency syndrome. Here we demonstrate that knockout (KO) of AP-4 in a mouse model leads to thinning of the corpus callosum and ventricular enlargement, anatomical defects previously described in patients. At the cellular level, we find that AP-4 KO leads to defects in axonal extension and branching, in addition to aberrant distal swellings. Interestingly, we show that ATG9A, a key protein in autophagosome maturation, is critically dependent on AP-4 for its sorting from the trans-golgi network. Failure of AP-4 mediated ATG9A sorting results in its dramatic retention in the trans-golgi network in vitro and in vivo leading to a specific reduction of the axonal pool of ATG9A. As a result, autophagosome biogenesis is aberrant in the axon of AP-4 deficient neurons. The specific alteration to axonal integrity and axonal autophagosome maturation in AP-4 knockout neurons may underpin the pathology of AP-4 deficiency.

AP-4 mediated ATG9A sorting underlies axonal and autophagosome biogenesis defects in a mouse model of AP-4 deficiency syndrome