Investigation of PTC124-mediated Translational Readthrough in a Retinal Organoid Model of AIPL1-associated Leber Congenital Amaurosis

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2022 Sep 9

PMID 36084639

Authors

Amy Leung

Almudena Sacristan-Reviriego

Pedro R L Perdigao

Hali Sai

Michalis Georgiou

Angelos Kalitzeos

Amanda-Jayne F Carr

Peter J Coffey

Michel Michaelides

James Bainbridge

Michael E Cheetham

Jacqueline van der Spuy

Affiliations

Soon will be listed here.

Abstract

Leber congenital amaurosis type 4 (LCA4), caused by AIPL1 mutations, is characterized by severe sight impairment in infancy and rapidly progressing degeneration of photoreceptor cells. We generated retinal organoids using induced pluripotent stem cells (iPSCs) from renal epithelial cells obtained from four children with AIPL1 nonsense mutations. iPSC-derived photoreceptors exhibited the molecular hallmarks of LCA4, including undetectable AIPL1 and rod cyclic guanosine monophosphate (cGMP) phosphodiesterase (PDE6) compared with control or CRISPR-corrected organoids. Increased levels of cGMP were detected. The translational readthrough-inducing drug (TRID) PTC124 was investigated as a potential therapeutic agent. LCA4 retinal organoids exhibited low levels of rescue of full-length AIPL1. However, this was insufficient to fully restore PDE6 in photoreceptors and reduce cGMP. LCA4 retinal organoids are a valuable platform for in vitro investigation of novel therapeutic agents.

Citing Articles

A Comparative Analysis of Models for AAV-Mediated Gene Therapy for Inherited Retinal Diseases.

Alsalloum A, Gornostal E, Mingaleva N, Pavlov R, Kuznetsova E, Antonova E Cells. 2024; 13(20.

PMID: 39451224 PMC: 11506034. DOI: 10.3390/cells13201706.

Retinal Organoids from Induced Pluripotent Stem Cells of Patients with Inherited Retinal Diseases: A Systematic Review.

Lee Y, Jo D Stem Cell Rev Rep. 2024; 21(1):167-197.

PMID: 39422807 PMC: 11762450. DOI: 10.1007/s12015-024-10802-7.

Limb Girdle Muscular Dystrophy Type 2B (LGMD2B): Diagnosis and Therapeutic Possibilities.

Poudel B, Fletcher S, Wilton S, Aung-Htut M Int J Mol Sci. 2024; 25(11).

PMID: 38891760 PMC: 11171558. DOI: 10.3390/ijms25115572.

Effective AAV-mediated gene replacement therapy in retinal organoids modeling AIPL1-associated LCA4.

Sai H, Ollington B, Rezek F, Chai N, Lane A, Georgiadis T Mol Ther Nucleic Acids. 2024; 35(1):102148.

PMID: 38439910 PMC: 10910061. DOI: 10.1016/j.omtn.2024.102148.

The Potential and Application of iPSCs in Gene and Cell Therapy for Retinopathies and Optic Neuropathies.

Lapshin E, Gershovich Y, Karabelsky A Acta Naturae. 2024; 15(4):56-64.

PMID: 38234607 PMC: 10790360. DOI: 10.32607/actanaturae.25454.

References

Li M, Andersson-Lendahl M, Sejersen T, Arner A . Muscle dysfunction and structural defects of dystrophin-null sapje mutant zebrafish larvae are rescued by ataluren treatment. FASEB J. 2013; 28(4):1593-9. DOI: 10.1096/fj.13-240044. View

Kirschman L, Kolandaivelu S, Frederick J, Dang L, Goldberg A, Baehr W . The Leber congenital amaurosis protein, AIPL1, is needed for the viability and functioning of cone photoreceptor cells. Hum Mol Genet. 2010; 19(6):1076-87. PMC: 2830831. DOI: 10.1093/hmg/ddp571. View

Moosajee M, Tracey-White D, Smart M, Weetall M, Torriano S, Kalatzis V . Functional rescue of REP1 following treatment with PTC124 and novel derivative PTC-414 in human choroideremia fibroblasts and the nonsense-mediated zebrafish model. Hum Mol Genet. 2016; 25(16):3416-3431. DOI: 10.1093/hmg/ddw184. View

Hug N, Longman D, Caceres J . Mechanism and regulation of the nonsense-mediated decay pathway. Nucleic Acids Res. 2016; 44(4):1483-95. PMC: 4770240. DOI: 10.1093/nar/gkw010. View

Du M, Liu X, Welch E, Hirawat S, Peltz S, Bedwell D . PTC124 is an orally bioavailable compound that promotes suppression of the human CFTR-G542X nonsense allele in a CF mouse model. Proc Natl Acad Sci U S A. 2008; 105(6):2064-9. PMC: 2538881. DOI: 10.1073/pnas.0711795105. View

Iribarne M, Nishiwaki Y, Nakamura S, Araragi M, Oguri E, Masai I . Aipl1 is required for cone photoreceptor function and survival through the stability of Pde6c and Gc3 in zebrafish. Sci Rep. 2017; 7:45962. PMC: 5381001. DOI: 10.1038/srep45962. View

Lukovic D, Castro A, Kaya K, Munezero D, Gieser L, Davo-Martinez C . Retinal Organoids derived from hiPSCs of an AIPL1-LCA Patient Maintain Cytoarchitecture despite Reduced levels of Mutant AIPL1. Sci Rep. 2020; 10(1):5426. PMC: 7096529. DOI: 10.1038/s41598-020-62047-2. View

van der Spuy J, Chapple J, Clark B, Luthert P, Sethi C, Cheetham M . The Leber congenital amaurosis gene product AIPL1 is localized exclusively in rod photoreceptors of the adult human retina. Hum Mol Genet. 2002; 11(7):823-31. DOI: 10.1093/hmg/11.7.823. View

Kolandaivelu S, Huang J, Hurley J, Ramamurthy V . AIPL1, a protein associated with childhood blindness, interacts with alpha-subunit of rod phosphodiesterase (PDE6) and is essential for its proper assembly. J Biol Chem. 2009; 284(45):30853-61. PMC: 2781484. DOI: 10.1074/jbc.M109.036780. View

10.

Sacristan-Reviriego A, van der Spuy J . The Leber Congenital Amaurosis-Linked Protein AIPL1 and Its Critical Role in Photoreceptors. Adv Exp Med Biol. 2018; 1074:381-386. DOI: 10.1007/978-3-319-75402-4_47. View

11.

Ng M, Li H, Ghelfi M, Goldman Y, Cooperman B . Ataluren and aminoglycosides stimulate read-through of nonsense codons by orthogonal mechanisms. Proc Natl Acad Sci U S A. 2021; 118(2). PMC: 7812769. DOI: 10.1073/pnas.2020599118. View

12.

Sohocki M, Bowne S, Sullivan L, Blackshaw S, Cepko C, Payne A . Mutations in a new photoreceptor-pineal gene on 17p cause Leber congenital amaurosis. Nat Genet. 1999; 24(1):79-83. PMC: 2581448. DOI: 10.1038/71732. View

13.

Goldmann T, Overlack N, Wolfrum U, Nagel-Wolfrum K . PTC124-mediated translational readthrough of a nonsense mutation causing Usher syndrome type 1C. Hum Gene Ther. 2011; 22(5):537-47. DOI: 10.1089/hum.2010.067. View

14.

Schwarz N, Carr A, Lane A, Moeller F, Chen L, Aguila M . Translational read-through of the RP2 Arg120stop mutation in patient iPSC-derived retinal pigment epithelium cells. Hum Mol Genet. 2014; 24(4):972-86. PMC: 4986549. DOI: 10.1093/hmg/ddu509. View

15.

Ramsden C, Nommiste B, R Lane A, Carr A, Powner M, Smart M . Rescue of the MERTK phagocytic defect in a human iPSC disease model using translational read-through inducing drugs. Sci Rep. 2017; 7(1):51. PMC: 5427915. DOI: 10.1038/s41598-017-00142-7. View

16.

Ramamurthy V, Niemi G, Reh T, Hurley J . Leber congenital amaurosis linked to AIPL1: a mouse model reveals destabilization of cGMP phosphodiesterase. Proc Natl Acad Sci U S A. 2004; 101(38):13897-902. PMC: 518850. DOI: 10.1073/pnas.0404197101. View

17.

Mort M, Ivanov D, Cooper D, Chuzhanova N . A meta-analysis of nonsense mutations causing human genetic disease. Hum Mutat. 2008; 29(8):1037-47. DOI: 10.1002/humu.20763. View

18.

Manuvakhova M, Keeling K, Bedwell D . Aminoglycoside antibiotics mediate context-dependent suppression of termination codons in a mammalian translation system. RNA. 2000; 6(7):1044-55. PMC: 1369979. DOI: 10.1017/s1355838200000716. View

19.

Torriano S, Erkilic N, Baux D, Cereso N, De Luca V, Meunier I . The effect of PTC124 on choroideremia fibroblasts and iPSC-derived RPE raises considerations for therapy. Sci Rep. 2018; 8(1):8234. PMC: 5974348. DOI: 10.1038/s41598-018-26481-7. View

20.

Dharmaraj S, Leroy B, Sohocki M, Koenekoop R, Perrault I, Anwar K . The phenotype of Leber congenital amaurosis in patients with AIPL1 mutations. Arch Ophthalmol. 2004; 122(7):1029-37. DOI: 10.1001/archopht.122.7.1029. View