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PreprintCited by 11Retinal Development and DisordersRNA Research and SplicingRetinal Diseases and Treatments

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Deciphering the Impact of PROM1 Alternative Splicing on Human Photoreceptor Development and Maturation

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Lako, Majlinda, Molina, Marina Moya, Dorgau, Birthe, Flood, Emily, Letteboer, Stef, Lorentzen, Esben, Coxhead, Jonathan, Smith, Graham, Roepman, Ronald, Nagaraja-Grellscheid, Sushma, Armstrong, Lyle·2024·Cell Death and Disease
DOI 10.21203/rs.3.rs-4348361/v1PMC PMC11445533

Preprint — published as 10.1038/s41419-024-07105-7

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Abstract

Conclusions: Importantly, the use of retinal organoids provides a valuable platform to study AS and unravel disease mechanisms in a more physiologically relevant context, opening avenues for further research and potential therapeutic interventions. Together our data indicate that cones may be more sensitive to PROM1 exon 4 skipping, corroborating the pathogenesis of late-onset mild maculopathy. Introduction: Alternative splicing (AS) is a crucial mechanism contributing to proteomic diversity, which is highly regulated in tissue- and development-specific patterns. Retinal tissue exhibits one of the highest levels of AS. In particular, photoreceptors have a distinctive AS pattern involving the inclusion of microexons not found in other cell types. PROM1 whose encoded protein Prominin-1 is located in photoreceptor outer segments (OSs), undergoes exon 4 inclusion from the 12 th post-conception week of human development through adulthood. Exon 4 skipping in PROM1 is associated with late-onset mild maculopathy, however its role in photoreceptor maturation and function is unknown. Methods: In this study retinal organoids, a valuable model system, were employed in combination with phosphorodiamidate morpholino oligos (PMOs) to assess the role of exon 4 AS in the development of human retina. Retinal organoids were treated with the PMOs for four weeks after which RT-PCR, western blotting and immunofluorescence analysis were performed to assess exon 4 exclusion and its impact on photoreceptors. The transcriptome of treated ROs was studied by bulk RNA-Seq. Results: Our data demonstrate that 55% skipping of PROM1 exon 4 resulted in decreased Prominin-1 expression by 40%, abnormal accumulation of cones in the basal side of the retinal organoids as well as detectable cone photoreceptor cilium defects. Transcriptomic and western blot analyses revealed decreased expression of cone, inner segment and connecting cilium basal body markers, increased expression of genes associated with stress response and the ubiquitin-proteasome system and downregulation of autophagy.

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Metadata

Authors
Lako, M., Molina, M. M., Dorgau, B., Flood, E., Letteboer, S., Lorentzen, E., Coxhead, J., Smith, G., Roepman, R., Nagaraja-Grellscheid, S., & Armstrong, L.
Affiliations
  • Newcastle University (GB)
  • Radboud University Medical Center (NL)
  • Aarhus University (DK)
  • Durham University
Corresponding author
Lako, Majlinda
ORCID
Venue
Cell Death and Disease
Publisher
Springer Science and Business Media LLC
Published
2024-05-24
Volume
15
Issue
10
Pages
721-721
ISSN
2041-4889 (ISSN-L)
DOI
10.21203/rs.3.rs-4348361/v1
PMC
PMC11445533
OpenAlex
W4403007768
Cited by
11
References
1
Citations by year
2026: 6 · 2025: 4 · 2024: 1
FWCI
1.50
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Death and Disease (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Durham Research Online (Durham University) (publishedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Radboud Repository (Radboud University) (submittedVersion)
Funders
  • Wellcome Trust
  • Retina UK
  • Newcastle University
  • European Commission (HORIZON2020, 861329)
  • National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC/C016106/1)
  • Medical Research Council (MR/R006237/1, MR/X008304/1, MR/T017503/1, MR/X001687/1)
  • Engineering and Physical Sciences Research Council (EP/Y031016/1)
  • Biotechnology and Biological Sciences Research Council (BB/T004460/1)
Topics
  • Health Sciences › Medicine › Ophthalmology › Retinal Development and Disorders
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
  • Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
Keywords
CD133, alternative splicing, photoreceptor development, retinal organoids, cone photoreceptors, connecting cilium, basal bodies, microexons, autophagy, ubiquitin-proteasome system, stress response, human retina
MeSH terms
AC133 Antigen, Exons, Humans, Organoids, Retina, Alternative Splicing, Photoreceptor Cells, Vertebrate

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