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Cited by 53Retinal Development and DisordersPluripotent Stem Cells ResearchRetinal Diseases and Treatments

Antioxidant and lipid supplementation improve the development of photoreceptor outer segments in pluripotent stem cell-derived retinal organoids

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West, Emma L., Majumder, Paromita, Naeem, Arifa, Fernando, Milan, O'Hara-Wright, Michelle, Lanning, Emily, Kloc, Magdalena, Ribeiro, Joana +10 more·2022·Stem Cell Reports
DOI 10.1016/j.stemcr.2022.02.019PMC PMC9023802
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Abstract

The generation of retinal organoids from human pluripotent stem cells (hPSC) is now a well-established process that in part recapitulates retinal development. However, hPSC-derived photoreceptors that exhibit well-organized outer segment structures have yet to be observed. To facilitate improved inherited retinal disease modeling, we determined conditions that would support outer segment development in maturing hPSC-derived photoreceptors. We established that the use of antioxidants and BSA-bound fatty acids promotes the formation of membranous outer segment-like structures. Using new protocols for hPSC-derived retinal organoid culture, we demonstrated improved outer segment formation for both rod and cone photoreceptors, including organized stacked discs. Using these enhanced conditions to generate iPSC-derived retinal organoids from patients with X-linked retinitis pigmentosa, we established robust cellular phenotypes that could be ameliorated following adeno-associated viral vector-mediated gene augmentation. These findings should aid both disease modeling and the development of therapeutic approaches for the treatment of photoreceptor disorders.

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  • hESC-derived retinal organoids contain all the major cell types in the retina.

    “Here, we used our well-established 2D/3D differentiation protocol to generate hESC-derived retinal organoids containing all the major cell types in the retina (SI Appendix, Fig. S6) (26).”

    From Characterization of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina (2024) · cites this paper as [26]

    The Results section ('Efficient generation of human PSC-derived photoreceptors bearing outer segment-like structures'; page unavailable) describes generating hESC-derived retinal organoids and documents their photoreceptors. The Discussion also identifies rod and cone photoreceptors. Our copy does not establish that these organoids contain all the major retinal cell types. (LLM, conf=0.83)

    Experimental — accuracy not yet measured

  • Overall transduction was determined using an arbitrary cut-off (Cas13bt3 read counts ≥ 10) based on previous experience with retinal organoids.

    “It should also be noted that overall transduction was determined using an arbitrary cut-off (Cas13bt3 read counts ≥ 10) based on our previous experience with the retinal organoids (26).”

    From Characterization of RNA editing and gene therapy with a compact CRISPR-Cas13 in the retina (2024) · cites this paper as [26]

    The Results section on AAV-mediated gene supplementation reports transduction of retinal organoids and estimates its efficiency using GFP-positive cells. It does not describe Cas13bt3 read counts or a threshold of ≥10, so it supports the cited prior experience with organoid transduction but not the stated cut-off. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 35

  1. Anderson R.E. O’Brien P.J. Wiegand R.D. Koutz C.A. Stinson A.M. Conservation of docosahexaenoic acid in the retina Adv. Exp. Med. Biol. 318 1992 285 294 1386176
  2. Agbaga M.-P. Merriman D.K. Brush R.S. Lydic T.A. Conley S.M. Naash M.I. Jackson S. Woods A.S. Reid G.E. Busik J.V. Differential composition of DHA and very-long-chain PUFAs in rod and cone photoreceptors J. Lipid Res. 59 2018 1586 1596 29986998 PMC6121944
  3. Aït-Ali N. Fridlich R. Millet-Puel G. Clérin E. Delalande F. Jaillard C. Blond F. Perrocheau L. Reichman S. Byrne L.C. Rod-derived cone viability factor promotes cone survival by stimulating aerobic glycolysis Cell 161 2015 817 832 25957687
  4. Arai E. Parmar V.M. Sahu B. Perusek L. Parmar T. Maeda A. Docosahexaenoic acid promotes differentiation of photoreceptor cells in three-dimensional neural retinas Neurosci. Res. 123 2017 1 7 28433627
  5. Bazan N.G. Gordon W.C. de Turco E.B.R. Docosahexaenoic acid uptake and metabolism in photoreceptors: retinal conservation by an efficient retinal pigment epithelial cell-mediated recycling process Adv. Exp. Med. Biol. 318 1992 295 306 1386177
  6. Boulton M. Dayhaw-Barker P. The role of the retinal pigment epithelium: topographical variation and ageing changes Eye (London, England) 15 2001 384 389 11450762
  7. Brooks M.J. Chen H.Y. Kelley R.A. Mondal A.K. Nagashima K. Val N.D. Li T. Chaitankar V. Swaroop A. Improved retinal organoid differentiation by modulating signaling pathways revealed by comparative transcriptome analyses with development in vivo Stem Cell Rep. 13 2019 891 905 PMC6895716 31631019
  8. Deng W.-L. Gao M.-L. Lei X.-L. Lv J.-N. Zhao H. He K.-W. Xia X.-X. Li L.-Y. Chen Y.-C. Li Y.-P. Gene correction reverses ciliopathy and photoreceptor loss in iPSC-derived retinal organoids from retinitis pigmentosa patients Stem Cell Rep. 10 2018 1267 1281 PMC5998840 29526738
  9. Gakovic M. Shu X. Kasioulis I. Carpanini S. Moraga I. Wright A.F. The role of RPGR in cilia formation and actin stability Hum. Mol. Genet. 20 2011 4840 4850 21933838
  10. Gao M.-L. Lei X.-L. Han F. He K.-W. Jin S.-Q. Zhang Y.-Y. Jin Z.-B. Patient-specific retinal organoids recapitulate disease features of late-onset retinitis pigmentosa Front. Cell Dev. Biol. 8 2020 128 32211407 PMC7068133
  11. Gonzalez-Cordero A. West E.L. Pearson R.A. Duran Y. Carvalho L.S. Chu C.J. Naeem A. Blackford S.J.I. Georgiadis A. Lakowski J. Photoreceptor precursors derived from three-dimensional embryonic stem cell cultures integrate and mature within adult degenerate retina Nat. Biotechnol. 31 2013 741 747 23873086 PMC3826328
  12. Gonzalez-Cordero A. Kruczek K. Naeem A. Fernando M. Kloc M. Ribeiro J. Goh D. Duran Y. Blackford S.J.I. Abelleira-Hervas L. Recapitulation of human retinal development from human pluripotent stem cells generates transplantable populations of cone photoreceptors Stem Cell Rep. 9 2017 820 837 PMC5599247 28844659
  13. Hong D.H. Yue G. Adamian M. Li T. Retinitis pigmentosa GTPase regulator (RPGRr)-interacting protein is stably associated with the photoreceptor ciliary axoneme and anchors RPGR to the connecting cilium J. Biol. Chem. 276 2001 12091 12099 11104772
  14. Hong D.-H. Pawlyk B.S. Adamian M. Sandberg M.A. Li T. A single, abbreviated RPGR-ORF15 variant reconstitutes RPGR function in vivo Invest. Ophthalmol. Vis. Sci. 46 2005 435 441 15671266
  15. Khani S.C. Pawlyk B.S. Bulgakov O.V. Kasperek E. Young J.E. Adamian M. Sun X. Smith A.J. Ali R.R. Li T. AAV-mediated expression targeting of rod and cone photoreceptors with a human rhodopsin Kinase promoter Invest. Ophth Vis. Sci. 48 2007 3954 3961 17724172
  16. Kruczek K. Cordero A.G. Goh D. Naeem A. Jonikas M. Blackford S.J.I. Kloc M. Duran Y. Georgiadis A. Sampson R.D. Differentiation and transplantation of embryonic stem cell-derived cone photoreceptors into a mouse model of end-stage retinal degeneration Stem Cell Rep. 8 2017 1659 1674 PMC5470175 28552606
  17. Lane A. Jovanovic K. Shortall C. Ottaviani D. Panes A.B. Schwarz N. Guarascio R. Hayes M.J. Palfi A. Chadderton N. Modeling and rescue of RP2 retinitis pigmentosa using iPSC-derived retinal organoids Stem Cell Rep. 15 2020 67 79 PMC7363745 32531192
  18. Lukovic D. Castro A.A. Kaya K.D. Munezero D. Gieser L. Davó-Martínez C. Corton M. Cuenca N. Swaroop A. Ramamurthy V. Retinal organoids derived from hiPSCs of an AIPL1-LCA patient maintain cytoarchitecture despite reduced levels of mutant AIPL1 Sci. Rep. 10 2020 5426 32214115 PMC7096529
  19. Megaw R. Abu-Arafeh H. Jungnickel M. Mellough C. Gurniak C. Witke W. Zhang W. Khanna H. Mill P. Dhillon B. Gelsolin dysfunction causes photoreceptor loss in induced pluripotent cell and animal retinitis pigmentosa models Nat. Commun. 8 2017 271 28814713 PMC5559447
  20. Neuringer M. Connor W.E. Petten C.V. Barstad L. Dietary omega-3 fatty acid deficiency and visual loss in infant rhesus monkeys J. Clin. Invest. 73 1984 272 276 6317716 PMC425011
  21. Nguyen L.N. Ma D. Shui G. Wong P. Cazenave-Gassiot A. Zhang X. Wenk M.R. Goh E.L.K. Silver D.L. Mfsd2a is a transporter for the essential omega-3 fatty acid docosahexaenoic acid Nature 509 2014 503 506 24828044
  22. O’Hara-Wright M. Cordero A.G. Retinal organoids: a window into human retinal development Development 147 2020 dev189746 33361444 PMC7774906
  23. Ovando-Roche P. West E.L. Branch M.J. Sampson R.D. Fernando M. Munro P. Georgiadis A. Rizzi M. Kloc M. Naeem A. Use of bioreactors for culturing human retinal organoids improves photoreceptor yields Stem Cell Res. Ther. 2018 1 14 29895313 PMC5998504
  24. Parfitt D.A. Lane A. Ramsden C.M. Carr A.-J.F. Munro P.M. Jovanovic K. Schwarz N. Kanuga N. Muthiah M.N. Hull S. Identification and correction of mechanisms underlying inherited blindness in human iPSC-derived optic cups Cell Stem Cell 18 2016 769 781 27151457 PMC4899423
  25. Pawlyk B.S. Bulgakov O.V. Sun X. Adamian M. Shu X. Smith A.J. Berson E.L. Ali R.R. Khani S. Wright A.F. Photoreceptor rescue by an abbreviated human RPGR gene in a murine model of X-linked retinitis pigmentosa Gene Ther. 23 2016 196 204 26348595 PMC4863462
  26. Rotstein N.P. Aveldaño M.I. Barrantes F.J. Roccamo A.M. Politi L.E. Apoptosis of retinal photoreceptors during development in vitro: protective effect of docosahexaenoic acid J. Neurochem. 69 1997 504 513 9231708
  27. Shindou H. Koso H. Sasaki J. Nakanishi H. Sagara H. Nakagawa K.M. Takahashi Y. Hishikawa D. Iizuka-Hishikawa Y. Tokumasu F. Docosahexaenoic acid preserves visual function by maintaining correct disc morphology in retinal photoreceptor cells J. Biol. Chem. 292 2017 12054 12064 28578316 PMC5519357
  28. Strauss O. The retinal pigment epithelium in visual function Physiol. Rev. 85 2005 845 881 15987797
  29. Tee J.J.L. Smith A.J. Hardcastle A.J. Michaelides M. RPGR-associated retinopathy: clinical features, molecular genetics, animal models and therapeutic options Br. J. Ophthalmol. 100 2016 1022 1027 26843488
  30. Wahlin K.J. Maruotti J.A. Sripathi S.R. Ball J. Angueyra J.M. Kim C. Grebe R. Li W. Jones B.W. Zack D.J. Photoreceptor outer segment-like structures in long-term 3D retinas from human pluripotent stem cells Sci. Rep. 7 2017 766 28396597 PMC5429674
  31. Wheeler T.G. Benolken R.M. Anderson R.E. Visual membranes: specificity of fatty acid precursors for the electrical response to illumination Science (New York, N.Y.) 188 1975 1312 1314 1145197
  32. Wu Z. Hiriyanna S. Qian H. Mookherjee S. Campos M.M. Gao C. Fariss R. Sieving P.A. Li T. Colosi P. A long-term efficacy study of gene replacement therapy for RPGR-associated retinal degeneration Hum. Mol. Genet. 24 2015 3956 3970 25877300 PMC4476444
  33. Young R.W. A difference between rods and cones in the renewal of outer segment protein Invest. Ophthalmol. 8 1969 222 231 5777484
  34. Zhong X. Gutierrez C. Xue T. Hampton C. Vergara M.N. Cao L.-H. Peters A. Park T.S. Zambidis E.T. Meyer J.S. Generation of three-dimensional retinal tissue with functional photoreceptors from human iPSCs Nat. Commun. 5 2014 4047 24915161 PMC4370190
  35. Zhou S. Flamier A. Abdouh M. Tétreault N. Barabino A. Wadhwa S. Bernier G. Differentiation of human embryonic stem cells into cone photoreceptors through simultaneous inhibition of BMP, TGFβ and Wnt signaling Development 142 2015 3294 3306 26443633

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Authors
West, E. L., Majumder, P., Naeem, A., Fernando, M., O'Hara-Wright, M., Lanning, E., Kloc, M., Ribeiro, J., Ovando-Roche, P., Shum, I. O., Jumbu, N., Sampson, R., Hayes, M., Bainbridge, J. W. B., Georgiadis, A., Smith, A. J., Gonzalez-Cordero, A., & Ali, R. R.
Affiliations
  • University College London (GB)
  • Moorfields Eye Hospital NHS Foundation Trust (GB)
Corresponding author
  • Gonzalez-Cordero, Anai
  • Ali, Robin R.
ORCID
Venue
Stem Cell Reports
Publisher
Elsevier BV
Published
2022-04
Volume
17
Issue
4
Pages
775-788
ISSN
2213-6711 (ISSN-L)
DOI
10.1016/j.stemcr.2022.02.019
PMC
PMC9023802
OpenAlex
W4221032109
Cited by
53
References
1
Citations by year
2026: 15 · 2025: 12 · 2024: 18 · 2023: 4 · 2022: 4
FWCI
3.27
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Reports (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • UCL Discovery (University College London) (submittedVersion)
Funders
Medical Research Council (MR/J004553/1, MR/M007871/1, MR/L012758/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
Keywords
retinal organoids, photoreceptor outer segments, human pluripotent stem cells, induced pluripotent stem cells, antioxidants, lipid supplementation, rod photoreceptors, cone photoreceptors, X-linked retinitis pigmentosa, retinal disease modeling, retinal development
MeSH terms
Antioxidants, Humans, Lipids, Organoids, Retina, Retinal Cone Photoreceptor Cells, Dietary Supplements, Pluripotent Stem Cells

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