TDP-43 regulates LC3ylation in neural tissue through ATG4B cryptic splicing inhibition.

1. Acta Neuropathol. 2024 Sep 21;148(1):45. doi: 10.1007/s00401-024-02780-4. TDP-43 regulates LC3ylation in neural tissue through ATG4B cryptic splicing inhibition. Torres P(1), Rico-Rios S(1), Ceron-Codorniu M(1), Santacreu-Vilaseca M(1), Seoane-Miraz D(2)(3), Jad Y(2)(3), Ayala V(1), Mariño G(4)(5)(6), Beltran M(7), Miralles MP(7), Andrés-Benito P(8)(9), Fernandez-Irigoyen J(10), Santamaria E(10), López-Otín C(4)(5)(11), Soler RM(7), Povedano M(8)(9), Ferrer I(9)(12)(13), Pamplona R(1), Wood MJA(2)(3), Varela MA(14)(15), Portero-Otin M(16). Author information: (1)Metabolic Pathophysiology Research Group, Department of Experimental Medicine, University of Lleida (UdL), Lleida Biomedical Research Institute (IRBLleida), 25198, Lleida, Spain. (2)Department of Paediatrics, Institute of Developmental and Regenerative Medicine (IDRM), University of Oxford, Roosevelt Dr, Oxford, OX3 7TY, UK. (3)MDUK Oxford Neuromuscular Centre, University of Oxford, Oxford, UK. (4)Departamento de Biología Funcional, Facultad de Medicina, Universidad de Oviedo, 33006, Oviedo, Spain. (5)Instituto Universitario de Oncología (IUOPA), 33006, Oviedo, Spain. (6)Instituto de Investigación Sanitaria Del Principado de Asturias (ISPA), 33011, Oviedo, Spain. (7)Neuronal Signaling Unit, Department of Experimental Medicine, Lleida Biomedical Research Institute (IRBLleida), University of Lleida (UdL), 25198, Lleida, Spain. (8)Neurologic Diseases and Neurogenetics Group, Bellvitge Institute for Biomedical Research (IDIBELL), 08907, L'Hospitalet de Llobregat, Spain. (9)CIBERNED (Network Centre of Biomedical Research of Neurodegenerative Diseases), Institute of Health Carlos III, 08907, L'Hospitalet de Llobregat, Barcelona, Spain. (10)Clinical Neuroproteomics Unit, Proteomics Platform, Proteored-ISCIII, Navarrabiomed, Complejo Hospitalario de Navarra (CHN), Universidad Pública de Navarra (UPNA), diSNA, 31008, Pamplona, Spain. (11)Departamento de Bioquímica y Biología Molecular, Universidad de Oviedo, 33006, Oviedo, Spain. (12)Neuropathology Group, Institute of Biomedical Research, IDIBELL, 08907, L'Hospitalet de Llobregat, Spain. (13)Department of Pathology and Experimental Therapeutics, University of Barcelona, 08007, Barcelona, Spain. (14)Department of Paediatrics, Institute of Developmental and Regenerative Medicine (IDRM), University of Oxford, Roosevelt Dr, Oxford, OX3 7TY, UK. miguel.varela@paediatrics.ox.ac.uk. (15)MDUK Oxford Neuromuscular Centre, University of Oxford, Oxford, UK. miguel.varela@paediatrics.ox.ac.uk. (16)Metabolic Pathophysiology Research Group, Department of Experimental Medicine, University of Lleida (UdL), Lleida Biomedical Research Institute (IRBLleida), 25198, Lleida, Spain. manuel.portero@udl.cat. 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. © 2024. The Author(s). DOI: 10.1007/s00401-024-02780-4 PMCID: PMC11416411 PMID: 39305312 [Indexed for MEDLINE] Conflict of interest statement: P.T., M.J.A.W., M.A.V., M.P.-O. are inventors in a patent application (2315883.5), priority request in UK covering the use of ASOs targeting ATG4B cryptic exon and multi-targets for TDP-43 proteinopathies.