A Human Retinal Pigment Epithelium-Based Screening Platform Reveals Inducers of Photoreceptor Outer Segments Phagocytosis
| العنوان: | A Human Retinal Pigment Epithelium-Based Screening Platform Reveals Inducers of Photoreceptor Outer Segments Phagocytosis |
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| المؤلفون: | Katerina Vafia, Marius Ader, Elly M. Tanaka, Rico Barsacchi, Mike O. Karl, Sven Schreiter, Seba Almedawar, Stephen H. Tsang, Marc Bickle |
| المصدر: | Stem Cell Reports Stem cell reports 15(6), 1347-1361 (2020). doi:10.1016/j.stemcr.2020.10.013 |
| بيانات النشر: | Elsevier BV, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, MERTK, metabolism [Human Embryonic Stem Cells], Phagocytosis, Human Embryonic Stem Cells, Retinal Pigment Epithelium, Biology, Biochemistry, Article, Cell Line, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, cytology [Human Embryonic Stem Cells], RP38, Retinitis pigmentosa, Genetics, medicine, Humans, ddc:610, metabolism [Photoreceptor Cells, Vertebrate], Induced pluripotent stem cell, cytology [Retinal Pigment Epithelium], Retinal pigment epithelium, cytology [Photoreceptor Cells, Vertebrate], phagocytosis, Retinal, Ramoplanin, Cell Biology, medicine.disease, Photoreceptor outer segment, Embryonic stem cell, eye diseases, POS, Cell biology, 030104 developmental biology, medicine.anatomical_structure, chemistry, RPE, sense organs, metabolism [Retinal Pigment Epithelium], 030217 neurology & neurosurgery, Photoreceptor Cells, Vertebrate, Developmental Biology |
| الوصف: | Summary Phagocytosis is a key function in various cells throughout the body. A deficiency in photoreceptor outer segment (POS) phagocytosis by the retinal pigment epithelium (RPE) causes vision loss in inherited retinal diseases and possibly age-related macular degeneration. To date, there are no effective therapies available aiming at recovering the lost phagocytosis function. Here, we developed a high-throughput screening assay based on RPE derived from human embryonic stem cells (hRPE) to reveal enhancers of POS phagocytosis. One of the hits, ramoplanin (RM), reproducibly enhanced POS phagocytosis and ensheathment in hRPE, and enhanced the expression of proteins known to regulate membrane dynamics and ensheathment in other cell systems. Additionally, RM rescued POS internalization defect in Mer receptor tyrosine kinase (MERTK) mutant hRPE, derived from retinitis pigmentosa patient induced pluripotent stem cells. Our platform, including a primary phenotypic screening phagocytosis assay together with orthogonal assays, establishes a basis for RPE-based therapy discovery aiming at a broad patient spectrum. Graphical Abstract Highlights • A phenotypic screening assay based on human RPE and POS was established • Screen identifies ramoplanin as an inducer of POS phagocytosis in hESC-RPE • Ramoplanin upregulates expression of genes involved in POS ensheathment • Ramoplanin rescues POS phagocytosis defect in retinitis pigmentosa RPE In this work, Schreiter et al. describe the establishment of a screening platform consisting of primary phenotypic assay and secondary assays based on hPSC-derived RPE. Using this platform ramoplanin was identified as a hit that upregulates POS phagocytosis in wild-type RPE and rescues phagocytosis defect in disease RPE. |
| تدمد: | 2213-6711 |
| DOI: | 10.1016/j.stemcr.2020.10.013 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::644644442a10dcf23eed5900b6fde9a9 https://doi.org/10.1016/j.stemcr.2020.10.013 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....644644442a10dcf23eed5900b6fde9a9 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 22136711 |
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| DOI: | 10.1016/j.stemcr.2020.10.013 |