HIWI2 Influences Endosomal Trafficking and Eph Receptor Signaling in Photoreceptor Cells

Photoreceptor integrity depends on the precise coordination of membrane trafficking and signal transmission. Despite their well-known roles in germline biology, the functions of PIWI family proteins in post-mitotic neuronal cells remain unclear. We investigated the role of HIWI2 in photoreceptor-derived 661W cells. Silencing of HIWI2 resulted in a significant decrease in the early endosomal marker, Rab5, and its effector EEA1, and reduced expression of the recycling endosome marker Rab11, indicating poor endosomal sorting and receptor recycling. In contrast, the marker for late endosomes, Rab7, was significantly upregulated, suggesting a shift toward degradative trafficking pathways, in line with increased receptor breakdown. These trafficking shifts led to the degradation of EphA2 and EphB2 receptors, as confirmed by a phospho-proteome receptor tyrosine kinase array and further supported by immunoblotting, and were accompanied by a compensatory increase in Akt phosphorylation. Furthermore, HIWI2 deficiency impaired cell motility in wound-healing assays. These results propose HIWI2 as a critical regulator of endosomal sorting and Eph receptor stability, providing a novel link between the PIWI pathway and photoreceptor integrity.