CRB1 is required for recycling by RAB11A+ vesicles in human retinal organoids

Thilo M. Buck, Peter M. J. Quinn, Lucie P. Pellissier, Aat A. Mulder, Aldo Jongejan, Xuefei Lu, Nanda Boon, Daniëlle Koot, Hind Almushattat, Christiaan H. Arendzen, Rogier M. Vos, Edward J. Bradley, Christian Freund, Harald M. M. Mikkers, Camiel J. F. Boon, Perry D. Moerland, Frank Baas, Abraham J. Koster, Jacques Neefjes, Ilana BerlinCarolina R. Jost, Jan Wijnholds

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)

Abstract

CRB1 gene mutations can cause early- or late-onset retinitis pigmentosa, Leber congenital amaurosis, or maculopathy. Recapitulating human CRB1 phenotypes in animal models has proven challenging, necessitating the development of alternatives. We generated human induced pluripotent stem cell (iPSC)-derived retinal organoids of patients with retinitis pigmentosa caused by biallelic CRB1 mutations and evaluated them against autologous gene-corrected hiPSCs and hiPSCs from healthy individuals. Patient organoids show decreased levels of CRB1 and NOTCH1 expression at the retinal outer limiting membrane. Proximity ligation assays show that human CRB1 and NOTCH1 can interact via their extracellular domains. CRB1 patient organoids feature increased levels of WDFY1+ vesicles, fewer RAB11A+ recycling endosomes, decreased VPS35 retromer complex components, and more degradative endolysosomal compartments relative to isogenic control organoids. Taken together, our data demonstrate that patient-derived retinal organoids enable modeling of retinal degeneration and highlight the importance of CRB1 in early endosome maturation receptor recycling in the retina.
Original languageEnglish
Pages (from-to)1793-1810
Number of pages18
JournalStem cell reports
Volume18
Issue number9
DOIs
Publication statusPublished - 12 Sept 2023

Keywords

  • CRB1
  • NOTCH1
  • RAB11A
  • VPS35
  • WDFY1
  • autophagy
  • cell polarity
  • endolysosomal system
  • organoids
  • retina
  • retromer

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