Identification and Correction of Mechanisms Underlying Inherited Blindness in Human iPSC-Derived Optic Cups

Cell Stem Cell. 2016 Jun 2;18(6):769-781. doi: 10.1016/j.stem.2016.03.021. Epub 2016 Apr 14.

Abstract

Leber congenital amaurosis (LCA) is an inherited retinal dystrophy that causes childhood blindness. Photoreceptors are especially sensitive to an intronic mutation in the cilia-related gene CEP290, which causes missplicing and premature termination, but the basis of this sensitivity is unclear. Here, we generated differentiated photoreceptors in three-dimensional optic cups and retinal pigment epithelium (RPE) from iPSCs with this common CEP290 mutation to investigate disease mechanisms and evaluate candidate therapies. iPSCs differentiated normally into RPE and optic cups, despite abnormal CEP290 splicing and cilia defects. The highest levels of aberrant splicing and cilia defects were observed in optic cups, explaining the retinal-specific manifestation of this CEP290 mutation. Treating optic cups with an antisense morpholino effectively blocked aberrant splicing and restored expression of full-length CEP290, restoring normal cilia-based protein trafficking. These results provide a mechanistic understanding of the retina-specific phenotypes in CEP290 LCA patients and potential strategies for therapeutic intervention.

MeSH terms

  • Antigens, Neoplasm / genetics
  • Antigens, Neoplasm / metabolism
  • Blindness / pathology*
  • Blindness / therapy*
  • Cell Cycle Proteins
  • Cell Differentiation / drug effects
  • Cilia / drug effects
  • Cilia / metabolism
  • Cytoskeletal Proteins
  • Exons / genetics
  • Eye Proteins / metabolism
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Inheritance Patterns / genetics*
  • Leber Congenital Amaurosis / pathology
  • Male
  • Morpholinos / pharmacology
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Opsins / metabolism
  • Optic Disk / cytology*
  • Organogenesis / drug effects
  • Photoreceptor Cells, Vertebrate / metabolism
  • Photoreceptor Cells, Vertebrate / pathology
  • Photoreceptor Cells, Vertebrate / ultrastructure
  • RNA Splicing / drug effects
  • RNA Splicing / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Retinal Pigment Epithelium / drug effects
  • Retinal Pigment Epithelium / metabolism
  • Retinal Pigment Epithelium / pathology
  • Retinal Pigment Epithelium / ultrastructure
  • rab GTP-Binding Proteins / metabolism

Substances

  • Antigens, Neoplasm
  • Cell Cycle Proteins
  • Cep290 protein, human
  • Cytoskeletal Proteins
  • Eye Proteins
  • Morpholinos
  • Neoplasm Proteins
  • Opsins
  • RNA, Messenger
  • RPGR protein, human
  • RAB8A protein, human
  • rab GTP-Binding Proteins