International Journal of Stem Cells

Korean Society for Stem Cell Research (한국줄기세포학회)
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Human Pluripotent Stem Cell-Derived Retinal Organoids: A Viable Platform for Investigating the Efficacy of Adeno-Associated Virus Gene Therapy

  • Hyeon-Jin Na (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jae-Eun Kwon (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seung-Hyun Kim (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jiwon Ahn (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Ok-Seon Kwon (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kyung-Sook Chung (Stem Cell Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Received : 2023.05.24
  • Accepted : 2023.11.28
  • Published : 2024.05.30
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Abstract

With recent advances in adeno-associated virus (AAV)-based gene therapy, efficacy and toxicity screening have become essential for developing gene therapeutic drugs for retinal diseases. Retinal organoids from human pluripotent stem cells (hPSCs) offer a more accessible and reproducible human test platform for evaluating AAV-based gene therapy. In this study, hPSCs were differentiated into retinal organoids composed of various types of retinal cells. The transduction efficiencies of AAV2 and AAV8, which are widely used in clinical trials of inherited retinal diseases, were analyzed using retinal organoids. These results suggest that retinal organoids derived from hPSCs serve as suitable screening platforms owing to their diverse retinal cell types and similarity to the human retina. In summary, we propose an optimal stepwise protocol that includes the generation of retinal organoids and analysis of AAV transduction efficacy, providing a comprehensive approach for evaluating AAV-based gene therapy for retinal diseases.

Keywords

Acknowledgement

This research was supported by grants from Korea Research Institute of Bioscience and Biotechnology (KRIBB) Research Initiative Program (KGM5362313), and the National Research Foundation of Korea (NRF-2022R1C1C1011202).

References

  1. Cideciyan AV, Hauswirth WW, Aleman TS, et al. Human RPE65 gene therapy for Leber congenital amaurosis: persistence of early visual improvements and safety at 1 year. Hum Gene Ther 2009;20:999-1004
  2. Cukras C, Wiley HE, Jeffrey BG, et al. Retinal AAV8-RS1 gene therapy for X-linked retinoschisis: initial findings from a phase I/IIa trial by intravitreal delivery. Mol Ther 2018;26:2282-2294 https://doi.org/10.1016/j.ymthe.2018.05.025
  3. Meng D, Ragi SD, Tsang SH. Therapy in rhodopsin-mediated autosomal dominant retinitis pigmentosa. Mol Ther 2022; 30:2633 Erratum for: Mol Ther 2020;28:2139-2149
  4. Wang D, Tai PWL, Gao G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat Rev Drug Discov 2019;18:358-378 https://doi.org/10.1038/s41573-019-0012-9
  5. Nakano T, Ando S, Takata N, et al. Self-formation of optic cups and storable stratified neural retina from human ESCs. Cell Stem Cell 2012;10:771-785 https://doi.org/10.1016/j.stem.2012.05.009
  6. Zhong X, Gutierrez C, Xue T, et al. Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs. Nat Commun 2014;5:4047
  7. Kuwahara A, Ozone C, Nakano T, Saito K, Eiraku M, Sasai Y. Generation of a ciliary margin-like stem cell niche from self-organizing human retinal tissue. Nat Commun 2015;6:6286
  8. Cowan CS, Renner M, De Gennaro M, et al. Cell types of the human retina and its organoids at single-cell resolution. Cell 2020;182:1623-1640.e34 https://doi.org/10.1016/j.cell.2020.08.013
  9. Finkbeiner C, Ortuno-Lizaran I, Sridhar A, Hooper M, Petter S, Reh TA. Single-cell ATAC-seq of fetal human retina and stem-cell-derived retinal organoids shows changing chromatin landscapes during cell fate acquisition. Cell Rep 2022;38:110294
  10. Saha A, Capowski E, Fernandez Zepeda MA, Nelson EC, Gamm DM, Sinha R. Cone photoreceptors in human stem cell-derived retinal organoids demonstrate intrinsic light responses that mimic those of primate fovea. Cell Stem Cell 2022;29:460-471.e3 Erratum in: Cell Stem Cell 2022;29:487-489 https://doi.org/10.1016/j.stem.2022.02.003
  11. Kwon OS, Na HJ, Ahn J, Chung KS. Establishment of a human induced pluripotent stem cell line, KRIBBi006-A, from peripheral blood mononuclear cells derived from a healthy male donor. Stem Cell Res 2022;65:102950
  12. Zou C, Levine EM. Vsx2 controls eye organogenesis and retinal progenitor identity via homeodomain and non-homeodomain residues required for high affinity DNA binding. PLoS Genet 2012;8:e1002924
  13. Xie H, Zhang W, Zhang M, et al. Chromatin accessibility analysis reveals regulatory dynamics of developing human retina and hiPSC-derived retinal organoids. Sci Adv 2020;6:eaay5247
  14. Garita-Hernandez M, Routet F, Guibbal L, et al. AAVmediated gene delivery to 3D retinal organoids derived from human induced pluripotent stem cells. Int J Mol Sci 2020;21:994
  15. Lane A, Jovanovic K, Shortall C, et al. Modeling and rescue of RP2 retinitis pigmentosa using iPSC-derived retinal organoids. Stem Cell Reports 2020;15:67-79 https://doi.org/10.1016/j.stemcr.2020.05.007
  16. Volkner M, Pavlou M, Buning H, Michalakis S, Karl MO. Optimized adeno-associated virus vectors for efficient transduction of human retinal organoids. Hum Gene Ther 2021;32:694-706 https://doi.org/10.1089/hum.2020.321
  17. Vandenberghe LH, Bell P, Maguire AM, et al. Dosage thresholds for AAV2 and AAV8 photoreceptor gene therapy in monkey. Sci Transl Med 2011;3:88ra54 Erratum in: Sci Transl Med 2011;3:112er9
  18. Surace EM, Auricchio A. Versatility of AAV vectors for retinal gene transfer. Vision Res 2008;48:353-359 https://doi.org/10.1016/j.visres.2007.07.027
  19. Achberger K, Cipriano M, Duchs MJ, et al. Human stem cell-based retina on chip as new translational model for validation of AAV retinal gene therapy vectors. Stem Cell Reports 2021;16:2242-2256
  20. Byrne LC, Day TP, Visel M, et al. In vivo-directed evolution of adeno-associated virus in the primate retina. JCI Insight 2020;5:e135112
  21. Cazier AP, Blazeck J. Advances in promoter engineering: novel applications and predefined transcriptional control. Biotechnol J 2021;16:e2100239
  22. Pan X, Yue Y, Boftsi M, et al. Rational engineering of a functional CpG-free ITR for AAV gene therapy. Gene Ther 2022;29:333-345
  23. Fligor CM, Lavekar SS, Harkin J, et al. Extension of retinofugal projections in an assembled model of human pluripotent stem cell-derived organoids. Stem Cell Reports 2021;16:2228-2241  https://doi.org/10.1016/j.stemcr.2021.05.009