WT1-MMP9 regulatory axis and its modulation by TNF-α in early and late onset glaucoma phenotypes

Glaucomas comprise a group of heterogeneous optic neuropathies that are linked to extracellular matrix (ECM) dysregulation, aberrant morphogenesis, and inflammatory signaling. Despite overlapping clinical features, the molecular underpinnings in early onset (primary congenital glaucoma [PCG]) and late onset (primary angle closure glaucoma [PACG]) diseases are poorly understood. Herein, we investigated the potential molecular connections between transcription factor WT1 and its downstream effector MMP9 along with TNF-α signaling towards bridging early and late-onset glaucomas. The study was driven by identifications of rare pathogenic variants in WT1 and MMP9 in a large cohort of PCG cases devoid of mutations in the candidate genes along with the significant association of a TNFα promoter variant in a cohort of PACG. Interestingly, the observed WT1 mutations clustered in the nuclear export sequence, uncovering a previously unrecognized WT1–MMP9 regulatory axis. WT1 is a versatile transcription factor located adjacent to PAX6, a master regulator of eye development. Here we showed combined wt1–pax6 suppression in zebrafish disrupted ocular morphogenesis, indicating a developmental interdependence. Further, WT1 acted as a repressor to MMP9 expression, in human trabecular meshwork (HTM) cells leading to reduced MMP9 function. We also demonstrated that TNFα signaling modulated this axis via nitric oxide (NO)–dependent nuclear export of WT1, leading to delayed MMP9 upregulation in HTM cells. This translocation was reversible by pharmacological inhibition of nuclear export or nitric oxide synthase, revealing a dynamic and drug-responsive mechanism. In addition, a patient-derived WT1 mutation (p.Pro264Leu) in PCG, impaired this translocation, leading to dysregulated MMP9 expression. Furthermore, LPS mediated inflammation in zebrafish resulted in elevated mmp9 and neurodegenerative markers, both mitigated by NO blockade, establishing the conservation of the TNFα–WT-MMP9 regulatory axis. Collectively, these findings suggested WT1 as a nodal regulator integrating developmental and inflammatory mechanisms in glaucoma pathogenesis. The shared transcriptional and signaling pathways expanded the mechanistic landscape across two distinct glaucoma phenotypes and identified potential molecular targets for early intervention and therapeutic modulation.