Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa

Nat Commun. 2018 Oct 12;9(1):4234. doi: 10.1038/s41467-018-06448-y.

Abstract

Mutations in pre-mRNA processing factors (PRPFs) cause autosomal-dominant retinitis pigmentosa (RP), but it is unclear why mutations in ubiquitously expressed genes cause non-syndromic retinal disease. Here, we generate transcriptome profiles from RP11 (PRPF31-mutated) patient-derived retinal organoids and retinal pigment epithelium (RPE), as well as Prpf31+/- mouse tissues, which revealed that disrupted alternative splicing occurred for specific splicing programmes. Mis-splicing of genes encoding pre-mRNA splicing proteins was limited to patient-specific retinal cells and Prpf31+/- mouse retinae and RPE. Mis-splicing of genes implicated in ciliogenesis and cellular adhesion was associated with severe RPE defects that include disrupted apical - basal polarity, reduced trans-epithelial resistance and phagocytic capacity, and decreased cilia length and incidence. Disrupted cilia morphology also occurred in patient-derived photoreceptors, associated with progressive degeneration and cellular stress. In situ gene editing of a pathogenic mutation rescued protein expression and key cellular phenotypes in RPE and photoreceptors, providing proof of concept for future therapeutic strategies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing / genetics
  • Alternative Splicing / physiology
  • Animals
  • Cell Adhesion / genetics
  • Cell Adhesion / physiology
  • Cell Differentiation / genetics
  • Cell Differentiation / physiology
  • Cilia / genetics
  • Cilia / metabolism
  • Cilia / physiology
  • Eye Proteins / genetics
  • Eye Proteins / metabolism*
  • Flow Cytometry
  • Humans
  • Immunohistochemistry
  • Induced Pluripotent Stem Cells / metabolism
  • Mice
  • Mutation / genetics
  • Organoids / cytology
  • Organoids / metabolism
  • RNA Splicing / genetics
  • RNA Splicing / physiology
  • Retina / cytology
  • Retina / metabolism
  • Retinitis Pigmentosa / etiology*
  • Retinitis Pigmentosa / genetics
  • Retinitis Pigmentosa / metabolism*

Substances

  • Eye Proteins
  • PRPF31 protein, mouse