RT info:eu-repo/semantics/article
T1 Clinical exome analysis and targeted gene repair of the c.1354dupT variant in iPSC lines from patients with PROM1-related retinopathies exhibiting diverse phenotypes
A1 Puertas Neyra, Kevin Louis
A1 Coco Martín, Rosa María
A1 Hernández Rodríguez, Leticia A
A1 Gobelli, Dino Joaquin
A1 García Ferrer, Yenisey
A1 Palma Vecino, Raicel
A1 Tellería Orriols, Juan José
A1 Simarro Grande, María
A1 Fuente García, Miguel Ángel de la
A1 Fernández Bueno, Iván
K1 iPSC, Retinal diseases, PROM1 gene, CD133, Retinitis pigmentosa, Cone-rod dystrophy, Stargardt’s type 4 disease
K1 3201.09 Oftalmología
AB ABSTRACT. BACKGROUND: Inherited retinal dystrophies (IRD) are one of the main causes of incurable blindness worldwide. IRD are caused by mutations in genes that encode essential proteins for the retina, leading to photoreceptor degeneration and loss of visual function. IRD generates an enormous global financial burden due to the lack of understanding of a significant part of its pathophysiology, molecular diagnosis, and the near absence of non-palliative treatment options. Patient-derived induced pluripotent stem cells (iPSC) for IRD seem to be an excellent option for addressing these questions, serving as exceptional tools for in-depth studies of IRD pathophysiology and testing new therapeutic approaches. METHODS: From a cohort of 8 patients with PROM1-related IRD, we identified 3 patients carrying the same variant (c.1354dupT) but expressing three different IRD phenotypes: Cone and rod dystrophy (CORD), Retinitis pigmentosa (RP), and Stargardt disease type 4 (STGD4). These three target patients, along with one healthy relative from each, underwent comprehensive ophthalmic examinations and their genetic panel study was expanded through clinical exome sequencing (CES). Subsequently, non-integrative patient-derived iPSC were generated and fully characterized. Correction of the c.1354dupT mutation was performed using CRISPR/Cas9, and the genetic restoration of the PROM1 gene was confirmed through flow cytometry and western blotting in the patient-derived iPSC lines. RESULTS: CES revealed that 2 target patients with the c.1354dupT mutation presented monoallelic variants in genes associated with the complement system or photoreceptor differentiation and peroxisome biogenesis disorders, respectively. The pluripotency and functionality of the patient-derived iPSC lines were confirmed, and the correction of the target mutation fully restored the capability of encoding Prominin-1 (CD133) in the genetically repaired patient-derived iPSC lines. CONCLUSIONS: The c.1354dupT mutation in the PROM1 gene is associated to three distinct AR phenotypes of IRD. This pleotropic effect might be related to the influence of monoallelic variants in other genes associated with retinal dystrophies. However, further evidence needs to be provided. Future experiments should include gene-edited patient-derived iPSC due to its potential as disease modelling tools to elucidate this matter in question.
PB BMC Part of Springer Nature
SN 1757-6512
YR 2024
FD 2024-07-02
LK https://uvadoc.uva.es/handle/10324/68995
UL https://uvadoc.uva.es/handle/10324/68995
LA eng
NO Stem Cell Res Ther. 2 Jul 2024, vol. 15, n. 1, Article number: 192, 24 páginas.
NO Producción Científica
DS UVaDOC
RD 21-nov-2024