MYO9A deficiency in motor neurons is associated with reduced neuromuscular agrin secretion

Hum Mol Genet. 2018 Apr 15;27(8):1434-1446. doi: 10.1093/hmg/ddy054.

Abstract

Congenital myasthenic syndromes (CMS) are a group of rare, inherited disorders characterized by compromised function of the neuromuscular junction, manifesting with fatigable muscle weakness. Mutations in MYO9A were previously identified as causative for CMS but the precise pathomechanism remained to be characterized. On the basis of the role of MYO9A as an actin-based molecular motor and as a negative regulator of RhoA, we hypothesized that loss of MYO9A may affect the neuronal cytoskeleton, leading to impaired intracellular transport. To investigate this, we used MYO9A-depleted NSC-34 cells (mouse motor neuron-derived cells), revealing altered expression of a number of cytoskeletal proteins important for neuron structure and intracellular transport. On the basis of these findings, the effect on protein transport was determined using a vesicular recycling assay which revealed impaired recycling of a neuronal growth factor receptor. In addition, an unbiased approach utilizing proteomic profiling of the secretome revealed a key role for defective intracellular transport affecting proper protein secretion in the pathophysiology of MYO9A-related CMS. This also led to the identification of agrin as being affected by the defective transport. Zebrafish with reduced MYO9A orthologue expression were treated with an artificial agrin compound, ameliorating defects in neurite extension and improving motility. In summary, loss of MYO9A affects the neuronal cytoskeleton and leads to impaired transport of proteins, including agrin, which may provide a new and unexpected treatment option.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actin Cytoskeleton / ultrastructure
  • Actins / genetics
  • Actins / metabolism
  • Agrin / genetics
  • Agrin / metabolism*
  • Amides
  • Animals
  • Cell Movement
  • Disease Models, Animal
  • Embryo, Nonmammalian
  • Enzyme Inhibitors
  • Gene Expression Regulation
  • Humans
  • Intermediate Filaments / genetics
  • Intermediate Filaments / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Motor Neurons / metabolism*
  • Motor Neurons / ultrastructure
  • Muscle Weakness / genetics*
  • Muscle Weakness / metabolism
  • Muscle Weakness / pathology
  • Myasthenic Syndromes, Congenital / genetics*
  • Myasthenic Syndromes, Congenital / metabolism
  • Myasthenic Syndromes, Congenital / pathology
  • Myosins / deficiency
  • Myosins / genetics*
  • Nerve Growth Factor / genetics*
  • Nerve Growth Factor / metabolism
  • Neuromuscular Junction / metabolism*
  • Neuromuscular Junction / ultrastructure
  • Protein Transport
  • Pyridines
  • Tubulin / genetics
  • Tubulin / metabolism
  • Zebrafish
  • rho GTP-Binding Proteins / genetics
  • rho GTP-Binding Proteins / metabolism

Substances

  • Actins
  • Agrin
  • Amides
  • Enzyme Inhibitors
  • Membrane Proteins
  • Pyridines
  • Tubulin
  • myosin IXa protein, mouse
  • periaxin
  • Y 27632
  • Nerve Growth Factor
  • Myosins
  • RhoA protein, mouse
  • rho GTP-Binding Proteins