title: Identification and Correction of Mechanisms Underlying Inherited Blindness in Human iPSC-Derived Optic Cups.
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.
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: David A
authors.initials: DA
authors.lastname: Parfitt
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Amelia
authors.initials: A
authors.lastname: Lane
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK; Moorfields Eye Hospital, 162 City Road, London EC1V 2PD, UK.
authors.forename: Conor M
authors.initials: CM
authors.lastname: Ramsden
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Amanda-Jayne F
authors.initials: AF
authors.lastname: Carr
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Munro
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Katarina
authors.initials: K
authors.lastname: Jovanovic
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Nele
authors.initials: N
authors.lastname: Schwarz
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Naheed
authors.initials: N
authors.lastname: Kanuga
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK; Moorfields Eye Hospital, 162 City Road, London EC1V 2PD, UK.
authors.forename: Manickam N
authors.initials: MN
authors.lastname: Muthiah
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK; Moorfields Eye Hospital, 162 City Road, London EC1V 2PD, UK.
authors.forename: Sarah
authors.initials: S
authors.lastname: Hull
authors.affiliation: Maurice Wohl Clinical Neurosciences Institute, Institute of Psychiatry, Psychology, and Neuroscience, Kings College London, London SE5 9NU, UK.
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Gallo
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK; Moorfields Eye Hospital, 162 City Road, London EC1V 2PD, UK.
authors.forename: Lyndon
authors.initials: L
authors.lastname: da Cruz
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK; Moorfields Eye Hospital, 162 City Road, London EC1V 2PD, UK.
authors.forename: Anthony T
authors.initials: AT
authors.lastname: Moore
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Alison J
authors.initials: AJ
authors.lastname: Hardcastle
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Coffey
authors.affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, London EC1V 9EL, UK. Electronic address: michael.cheetham@ucl.ac.uk.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Cheetham
mesh_terms: D000951:Antigens, Neoplasm / Q000235:genetics / Q000378:metabolism; D001766:Blindness / Q000473:pathology* / Q000628:therapy*; D018797:Cell Cycle Proteins; D002454:Cell Differentiation / Q000187:drug effects; D002923:Cilia / Q000187:drug effects / Q000378:metabolism; D003598:Cytoskeletal Proteins; D005091:Exons / Q000235:genetics; D005136:Eye Proteins / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D040582:Inheritance Patterns / Q000235:genetics*; D057130:Leber Congenital Amaurosis / Q000473:pathology; D008297:Male; D060172:Morpholinos / Q000494:pharmacology; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D055355:Opsins / Q000378:metabolism; D009898:Optic Disk / Q000166:cytology*; D038081:Organogenesis / Q000187:drug effects; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D012326:RNA Splicing / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D020691:rab GTP-Binding Proteins / Q000378:metabolism
chemical_list: D000951:Antigens, Neoplasm; D018797:Cell Cycle Proteins; C493903:Cep290 protein, human; D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D060172:Morpholinos; D009363:Neoplasm Proteins; D055355:Opsins; D012333:RNA, Messenger; C099929:RPGR protein, human; C082377:RAB8A protein, human; D020691:rab GTP-Binding Proteins