BUSCADOR RECOLECTA

Amyotrophic lateral sclerosis (ALS) is an adult-onset motor neuron disease with a mean survival time of three years. The 97% of the cases have TDP-43 nuclear depletion and cytoplasmic aggregation in motor neurons. TDP-43 prevents non-conserved cryptic exon splicing in certain genes, maintaining transcript stability, including ATG4B, which is crucial for autophagosome maturation and Microtubule-associated proteins 1A/1B light chain 3B (LC3B) homeostasis. In ALS mice (G93A), Atg4b depletion worsens survival rates and autophagy function. For the first time, we observed an elevation of LC3ylation in the CNS of both ALS patients and atg4b−/− mouse spinal cords. Furthermore, LC3ylation modulates the distribution of ATG3 across membrane compartments. Antisense oligonucleotides (ASOs) targeting cryptic exon restore ATG4B mRNA in TARDBP knockdown cells. We further developed multi-target ASOs targeting TDP-43 binding sequences for a broader effect. Importantly, our ASO based in peptide-PMO conjugates show brain distribution post-IV administration, offering a non-invasive ASO-based treatment avenue for neurodegenerative diseases., Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. G.M. was supported by grants from Ministerio Ciencia e Innovación (Spain) (PID2021-127534OB-I00). M.P.-O. and R.M.S. were supported by grants from Instituto de Salud Carlos III (PI20/000155, PI23/00176 and PI20/00098). R.P. was supported by Generalitat de Catalunya (2021 SGR 00990). P.T. was supported by an Unzué-Luzón grant. P.T. received a Margarita Salas postdoctoral fellowship from the Spanish Ministry of Universities (Spanish Government) which was supported by NextGenerationEU. Work supported by IRBLleida Scientific and Technical Service of Immunohistochemistry.