Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug
Abstract Background The most common gene responsible for autosomal recessive retinitis pigmentosa (RP) is EYS. The manner of decay of genetically defective EYS gene transcripts varies depending on the type of mutation using our cellular model, which consists of induced photoreceptor-directed fibrobl...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-04-01
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| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02827-x |
| _version_ | 1828378324659863552 |
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| author | Dilip Rai Masaki Iwanami Yoriko Takahashi Yukari Komuta Noriyuki Aoi Akihiro Umezawa Yuko Seko |
| author_facet | Dilip Rai Masaki Iwanami Yoriko Takahashi Yukari Komuta Noriyuki Aoi Akihiro Umezawa Yuko Seko |
| author_sort | Dilip Rai |
| collection | DOAJ |
| description | Abstract Background The most common gene responsible for autosomal recessive retinitis pigmentosa (RP) is EYS. The manner of decay of genetically defective EYS gene transcripts varies depending on the type of mutation using our cellular model, which consists of induced photoreceptor-directed fibroblasts from EYS-RP patients (EYS-RP cells). However, disease-specific profiles have not been clarified in EYS-RP cells. Herein we investigated comprehensive gene expression patterns and restoration of altered expression by low molecular weight molecules in EYS-RP cells. Methods Using induced photoreceptor-like cells by CRX, RAX, NeuroD, and OTX2, we employed qRT-PCR and DNA microarray analysis to compare expression levels of disease-related genes in EYS-RP cells. We investigated the effect of antiapoptotic or anti-endoplasmic reticulum (ER) stress/antioxidant reagents on the restoration of altered gene expression. Results Expression levels of phototransduction-related genes (blue opsin, rhodopsin, S-antigen, GNAT1, GNAT2) were lower in EYS-RP cells. CRYGD was extracted by global gene expression analysis, as a downregulated, retina-related and apoptosis-, endoplasmic reticulum (ER) stress- or aging-related gene. Pathway enrichment analysis suggested that “complement and coagulation cascades,” “ECM-receptor interaction” and “PI3K-Akt signaling pathway” could be involved in EYS-RP-associated pathogenesis. Among the matching/overlapping genes involved in those pathways, F2R was suggested as an EYS-RP-associated gene. The downregulation of CRYGD and F2R was completely restored by additional 4-PBA, an inhibitor of ER stress, and partially restored by metformin or NAC. In addition, 4-PBA normalized the expression level of cleaved caspase-3. Conclusions Our cellular model may reflect the ER stress-mediated degenerative retina and serve as a pathogenesis-oriented cost-effective rescue strategy for RP patients. |
| first_indexed | 2024-04-14T08:25:27Z |
| format | Article |
| id | doaj.art-797a7297b0fc4f06b589b5c8a7cda1e5 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-04-14T08:25:27Z |
| publishDate | 2022-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-797a7297b0fc4f06b589b5c8a7cda1e52022-12-22T02:04:06ZengBMCStem Cell Research & Therapy1757-65122022-04-0113111810.1186/s13287-022-02827-xEvaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drugDilip Rai0Masaki Iwanami1Yoriko Takahashi2Yukari Komuta3Noriyuki Aoi4Akihiro Umezawa5Yuko Seko6Sensory Functions Section, Research Institute, National Rehabilitation Center for Persons With DisabilitiesDepartment of Ophthalmology, Hospital, National Rehabilitation Center for Persons With DisabilitiesBioscience and Healthcare Engineering Division, Mitsui Knowledge Industry Co., Ltd.Sensory Functions Section, Research Institute, National Rehabilitation Center for Persons With DisabilitiesDepartment of Plastic, Oral and Maxillofacial Surgery, Teikyo University School of MedicineNational Center for Child Health and Development, Research InstituteSensory Functions Section, Research Institute, National Rehabilitation Center for Persons With DisabilitiesAbstract Background The most common gene responsible for autosomal recessive retinitis pigmentosa (RP) is EYS. The manner of decay of genetically defective EYS gene transcripts varies depending on the type of mutation using our cellular model, which consists of induced photoreceptor-directed fibroblasts from EYS-RP patients (EYS-RP cells). However, disease-specific profiles have not been clarified in EYS-RP cells. Herein we investigated comprehensive gene expression patterns and restoration of altered expression by low molecular weight molecules in EYS-RP cells. Methods Using induced photoreceptor-like cells by CRX, RAX, NeuroD, and OTX2, we employed qRT-PCR and DNA microarray analysis to compare expression levels of disease-related genes in EYS-RP cells. We investigated the effect of antiapoptotic or anti-endoplasmic reticulum (ER) stress/antioxidant reagents on the restoration of altered gene expression. Results Expression levels of phototransduction-related genes (blue opsin, rhodopsin, S-antigen, GNAT1, GNAT2) were lower in EYS-RP cells. CRYGD was extracted by global gene expression analysis, as a downregulated, retina-related and apoptosis-, endoplasmic reticulum (ER) stress- or aging-related gene. Pathway enrichment analysis suggested that “complement and coagulation cascades,” “ECM-receptor interaction” and “PI3K-Akt signaling pathway” could be involved in EYS-RP-associated pathogenesis. Among the matching/overlapping genes involved in those pathways, F2R was suggested as an EYS-RP-associated gene. The downregulation of CRYGD and F2R was completely restored by additional 4-PBA, an inhibitor of ER stress, and partially restored by metformin or NAC. In addition, 4-PBA normalized the expression level of cleaved caspase-3. Conclusions Our cellular model may reflect the ER stress-mediated degenerative retina and serve as a pathogenesis-oriented cost-effective rescue strategy for RP patients.https://doi.org/10.1186/s13287-022-02827-xRedirect differentiationDermal fibroblastPhotoreceptorDisease modelingRetinitis pigmentosaEYS |
| spellingShingle | Dilip Rai Masaki Iwanami Yoriko Takahashi Yukari Komuta Noriyuki Aoi Akihiro Umezawa Yuko Seko Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug Stem Cell Research & Therapy Redirect differentiation Dermal fibroblast Photoreceptor Disease modeling Retinitis pigmentosa EYS |
| title | Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug |
| title_full | Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug |
| title_fullStr | Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug |
| title_full_unstemmed | Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug |
| title_short | Evaluation of photoreceptor-directed fibroblasts derived from retinitis pigmentosa patients with defects in the EYS gene: a possible cost-effective cellular model for mechanism-oriented drug |
| title_sort | evaluation of photoreceptor directed fibroblasts derived from retinitis pigmentosa patients with defects in the eys gene a possible cost effective cellular model for mechanism oriented drug |
| topic | Redirect differentiation Dermal fibroblast Photoreceptor Disease modeling Retinitis pigmentosa EYS |
| url | https://doi.org/10.1186/s13287-022-02827-x |
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