| Resum: |
Injuries affecting peripheral or central nerves often lead to major deficits in motor and sensory function, underscoring their profound impact on patients' lives. Existing therapeutic strategies include non-surgical approaches, pharmacological approaches, administration of growth factors, and surgical interventions. However, these strategies still fail to address the complex regeneration environment in the Peripheral and Central Nervous Systems (PNS and CNS), and no effective approach is available for nervous injuries affecting both systems. Thus, gene therapy is a promising approach due to its cell-type specificity, ability to provide long-term effects without constant readministration, and less invasive surgeries. In this context, the anti-aging protein α-Klotho presents as a strong candidate given its multifaceted effects on neuroinflammation, oxidative stress, apoptosis, and myelination, processes central to nerve regeneration. This work has aimed to evaluate the effect of α-Klotho in degeneration, regeneration, and remyelination in the PNS, and to test an engineered AAV2 capsid for future application in optic nerve degeneration. Here, we have analyzed endogenous α-Klotho expression at the mRNA level after sciatic nerve resection and crush as distinct models for peripheral nerve injury (PNI), highlighting temporal and tissue-dependent differential regulation. In vitro, evaluating AAV-mediated sKL overexpression, we have detected a significant promotion in neuritogenesis in rat DRG organotypic cultures. In vivo, sKL overexpression through gene therapy with intrathecal administration enhanced axonal regeneration and remyelination in the tibial nerve after sciatic nerve cut-and-paste. With a view to its application in optic nerve atrophy, we have assessed the tropism of an engineered AAV2. GL capsid with different promoters, finding that under the control of the CAG promoter, higher retinal neuronal tropism was achieved than with the CMV promoter. Our results suggest that AAV-mediated α-Klotho overexpression improves nerve regeneration and remyelination, with potential clinical applications in PNI and optic nerve degeneration. |
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