Loss of CLN3, the gene mutated in juvenile neuronal ceroid lipofuscinosis, leads to metabolic impairment and autophagy induction in retinal pigment epithelium

Biochim Biophys Acta Mol Basis Dis. 2020 Oct 1;1866(10):165883. doi: 10.1016/j.bbadis.2020.165883. Epub 2020 Jun 25.


Juvenile neuronal ceroid lipofuscinosis (JNCL, aka. juvenile Batten disease or CLN3 disease) is a lysosomal storage disease characterized by progressive blindness, seizures, cognitive and motor failures, and premature death. JNCL is caused by mutations in the Ceroid Lipofuscinosis, Neuronal 3 (CLN3) gene, whose function is unclear. Although traditionally considered a neurodegenerative disease, CLN3 disease displays eye-specific effects: Vision loss not only is often one of the earliest symptoms of JNCL, but also has been reported in non-syndromic CLN3 disease. Here we described the roles of CLN3 protein in maintaining healthy retinal pigment epithelium (RPE) and normal vision. Using electroretinogram, fundoscopy and microscopy, we showed impaired visual function, retinal autofluorescent lesions, and RPE disintegration and metaplasia/hyperplasia in a Cln3 ~ 1 kb-deletion mouse model [1] on C57BL/6J background. Utilizing a combination of biochemical analyses, RNA-Seq, Seahorse XF bioenergetic analysis, and Stable Isotope Resolved Metabolomics (SIRM), we further demonstrated that loss of CLN3 increased autophagic flux, suppressed mTORC1 and Akt activities, enhanced AMPK activity, and up-regulated gene expression of the autophagy-lysosomal system in RPE-1 cells, suggesting autophagy induction. This CLN3 deficiency induced autophagy induction coincided with decreased mitochondrial oxygen consumption, glycolysis, the tricarboxylic acid (TCA) cycle, and ATP production. We also reported for the first time that loss of CLN3 led to glycogen accumulation despite of impaired glycogen synthesis. Our comprehensive analyses shed light on how loss of CLN3 affect autophagy and metabolism. This work suggests possible links among metabolic impairment, autophagy induction and lysosomal storage, as well as between RPE atrophy/degeneration and vision loss in JNCL.

Keywords: Autophagy; CLN3; Glycogen; Metabolism; Retinal pigment epithelium; Vision loss.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Atrophy / genetics
  • Atrophy / pathology
  • Autophagy
  • Blindness / genetics*
  • Blindness / pathology
  • Cell Line
  • Disease Models, Animal
  • Gene Knock-In Techniques
  • Gene Knockdown Techniques
  • Glycogen / metabolism
  • Humans
  • Lysosomes / pathology
  • Membrane Glycoproteins / deficiency*
  • Membrane Glycoproteins / genetics
  • Mice
  • Mice, Transgenic
  • Microscopy, Electron
  • Molecular Chaperones / genetics
  • Mutation
  • Neuronal Ceroid-Lipofuscinoses / complications
  • Neuronal Ceroid-Lipofuscinoses / genetics*
  • Neuronal Ceroid-Lipofuscinoses / pathology
  • RNA, Small Interfering / metabolism
  • Retinal Pigment Epithelium / pathology*
  • Retinal Pigment Epithelium / ultrastructure


  • CLN3 protein, human
  • CLN3 protein, mouse
  • Membrane Glycoproteins
  • Molecular Chaperones
  • RNA, Small Interfering
  • Glycogen