PUBLICATION

Unconventional secretory pathway activation restores hair cell mechanotransduction in an USH3A model

Authors
Gopal, S.R., Lee, Y.T., Stepanyan, R., McDermott, B.M., Alagramam, K.N.
ID
ZDB-PUB-190518-5
Date
2019
Source
Proceedings of the National Academy of Sciences of the United States of America   116(22): 11000-11009 (Journal)
Registered Authors
Alagramam, Kumar N., Gopal, Suhasini R., McDermott Jr., Brian M.
Keywords
USH3A, hearing loss, mitigation
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Artemisinins/pharmacology
  • Hair Cells, Auditory/drug effects
  • Hair Cells, Auditory/physiology*
  • Mechanotransduction, Cellular/genetics*
  • Membrane Proteins*/genetics
  • Membrane Proteins*/metabolism
  • Membrane Proteins*/physiology
  • Secretory Pathway/genetics*
  • Zebrafish
PubMed
31097578 Full text @ Proc. Natl. Acad. Sci. USA
Abstract
The pathogenic variant c.144T>G (p.N48K) in the clarin1 gene (CLRN1) results in progressive loss of vision and hearing in Usher syndrome IIIA (USH3A) patients. CLRN1 is predicted to be an essential protein in hair bundles, the mechanosensory structure of hair cells critical for hearing and balance. When expressed in animal models, CLRN1 localizes to the hair bundle, whereas glycosylation-deficient CLRN1N48K aggregates in the endoplasmic reticulum, with only a fraction reaching the bundle. We hypothesized that the small amount of CLRN1N48K that reaches the hair bundle does so via an unconventional secretory pathway and that activation of this pathway could be therapeutic. Using genetic and pharmacological approaches, we find that clarin1 knockout (clrn1KO/KO ) zebrafish that express the CLRN1c.144T>G pathogenic variant display progressive hair cell dysfunction, and that CLRN1N48K is trafficked to the hair bundle via the GRASP55 cargo-dependent unconventional secretory pathway (GCUSP). On expression of GRASP55 mRNA, or on exposure to the drug artemisinin (which activates GCUSP), the localization of CLRN1N48K to the hair bundles was enhanced. Artemisinin treatment also effectively restored hair cell mechanotransduction and attenuated progressive hair cell dysfunction in clrn1KO/KO larvae that express CLRN1c.144T>G , highlighting the potential of artemisinin to prevent sensory loss in CLRN1c.144T>G patients.
Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping