Effective treatment of optic neuropathies by intraocular delivery of MSC-sEVs through augmenting the G-CSF- macrophage pathway

Optic neuropathies, characterized by injury of retinal ganglion cell (RGC) axons of the optic nerve, cause incurable blindness worldwide. Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEVs) represent a promising "cell - free" therapy for regenerative medicine; however, the therapeutic effect on neural restoration fluctuates, and the underlying mechanism is poorly understood. Here, we illustrated that intraocular administration of MSC-sEVs promoted both RGC survival and axon regeneration in an optic nerve crush mouse model. Mechanistically, MSC-sEVs primarily targeted retinal mural cells to release high levels of colony- stimulating factor 3 (G-CSF) that recruited a neural restorative population of Ly6Clow monocytes/monocyte- derived macrophages (Mo/M(13). Intravitreal administration of G-CSF, a clinically proven agent for treating neutropenia, or donor Ly6Clow Mo/M(13 markedly improved neurological outcomes in vivo. Together, our data define a unique mechanism of MSC-sEV- induced G- CSF- to- Ly6Clow Mo/M(13 signaling in repairing optic nerve injury and highlight local delivery of MSC-sEVs, G-CSF, and Ly6Clow Mo/M(13 as therapeutic paradigms for the treatment of optic neuropathies.