From Injury Canalization to Molecular Rescue: A Human-Derived Retinal Ganglion Cell State Space Predicts Optic Nerve Injury and Treatment Response

Abstract

Background: Retinal ganglion cell (RGC) injury is typically analyzed by cell type, differential expression, or treatment-specificregeneration programs. A complementary question is whether injury and therapeutic response occupy a common molecularstate space that can be specified independently of the treatment dataset. Methods: A three-dimensional human RGC staterepresentation was defined by maintenance loss, stress response, and apoptosis-vulnerability coordinates. A pathologydirection was fixed from a human RGC branch architecture before evaluation of independent mouse data. First, 17,887 controland 13,906 optic-nerve-crush (ONC) day-2 RGCs from GSE137398 were used as an external injury validation. Second, 62,808RGCs from GSE201254 were analyzed across NoCrush, untreated ONC day 7, Pten, Pten+CNTF, and Pten/Socs3+CNTFgroups. A fixed 63-gene homologous panel was used without outcome-driven axis refitting. Results: In the external ONC day-2test, the pathology-score median increased from 0.258 in control to 1.855 after ONC, with a median shift of +1.598 (bootstrap95% CI 1.561-1.635) and rank-biserial effect 0.763. In GSE201254, median pathology score increased from 0.064 in NoCrushto 2.848 after untreated ONC day 7. Pten, Pten+CNTF, and Pten/Socs3+CNTF shifted medians to 0.757, 1.190, and 0.719,respectively; treatment-minus-ONC shifts were -2.091, -1.658, and -2.129 with bootstrap intervals excluding zero. Advancedinjury-state occupancy fell from 25.0% in untreated ONC to 3.0-4.9% after intervention, whereas 72.2-78.7% of treated RGCsoccupied the homeostatic-like region. Conclusions: A human-derived molecular direction generalized to independent mouseaxonal injury and showed an opposite redistribution under established neuroprotective/regenerative interventions. The resultssupport a treatment-responsive RGC molecular state-space hypothesis and identify an intermediate stress-dominant region asa candidate intervention window for causal testing. The analysis does not demonstrate reversal of individual-cell trajectories orclinical treatability. Keywords: retinal ganglion cell; optic nerve crush; single-cell RNA sequencing; molecular state space; PTEN; SOCS3; CNTF;neuroprotection; axon regeneration; therapeutic window