Generation and characterization of human induced pluripotent stem cells (iPSCs) from three patients with age-related macular degeneration carrying rare variants in the CFH gene

Koolen, Louet; Gagliardi, Giuliana; Brink, Sofie C.A. ten; Breuk, Anita de; Heesterbeek, Thomas J.; Hoyng, Carel B.; Albert, Silvia; Hollander, Anneke I. den

doi:10.1016/j.scr.2022.102669

Cited by 4 publications

(3 citation statements)

References 2 publications

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“…Blood was obtained from subjects arRP36 (heterozygous for c.4397-3890A>G), USH46 (heterozygous for c.1551-504C>T), and a healthy control. Peripheral blood mononuclear cells were isolated and reprogrammed into induced pluripotent stem cells (iPSCs) through episomal nucleofection, as described previously, 28 and subsequently differentiated into photoreceptor precursor cells (PPCs). 17 , 29 iPSC lines were generated at the Stem Cell Technology Center of the Radboudumc (Nijmegen, the Netherlands).…”

Section: Methodsmentioning

confidence: 99%

Whole genome sequencing for USH2A-associated disease reveals several pathogenic deep-intronic variants that are amenable to splice correction

Reurink¹,

Weisschuh²,

Garanto³

et al. 2023

Human Genetics and Genomics Advances

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Section: Methodsmentioning

confidence: 99%

Whole genome sequencing for USH2A-associated disease reveals several pathogenic deep-intronic variants that are amenable to splice correction

Reurink¹,

Weisschuh²,

Garanto³

et al. 2023

Human Genetics and Genomics Advances

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“…The generation of the patient-derived iPSC line from blood samples was performed by the Stem Cell Technology Center of the Radboud University Medical Center as described in previous publications. 33 – 37 One patient (patient B:II-1) harboring the complex allele c.[3023-15T>A;3023G>A] in IMPG2 and one control line were used to obtain PPCs as previously described 33 – 35 , 38 (see Supplementary Methods ). Two wells for each cell line were differentiated in each replicate.…”

Section: Methodsmentioning

confidence: 99%

Identification of a Complex Allele in IMPG2 as a Cause of Adult-Onset Vitelliform Macular Dystrophy

Vázquez-Domínguez

Fadaie

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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Purpose Inherited retinal diseases are a group of clinically and genetically heterogeneous disorders with approximately 270 genes involved. IMPG2 is associated with adult-onset vitelliform macular dystrophy. Here, we investigated two unrelated patients with vitelliform macular dystrophy to identify the underlying genetic cause. Methods Whole-exome sequencing identified a putative causal complex allele consisting of c.3023-15T>A and c.3023G>A (p.(Gly1008Asp)) in IMPG2 in both individuals. To assess its effect, in vitro splice assays in HEK293T and further characterization in patient-derived photoreceptor precursor cells (PPCs) were conducted. Results The results of the midigene splice assays in HEK293T showed that the complex allele causes a variety of splicing defects ranging from a small deletion to (multiple-)exon skipping. This finding was further validated using patient-derived PPCs that showed a significant increase of out-of-frame transcripts lacking one or multiple exons compared to control-derived PPCs. Overall, control PPCs consistently showed low levels of aberrantly spliced IMPG2 transcripts that were highly elevated in patient-derived PPCs. These differences were even more obvious upon inhibition of nonsense-mediated decay with cycloheximide. Conclusions We report a heterozygous complex allele in IMPG2 causative for adult-onset vitelliform macular dystrophy in two unrelated individuals with mild visual loss and bilateral vitelliform lesions. The predicted causal missense mutation c.3023G>A, located in the consensus splice acceptor site, enhances the splicing effect of the upstream variant c.3023-15T>A, leading to the generation of aberrant transcripts that decrease the full-length IMPG2 levels. These results suggest a haploinsufficiency mechanism of action and highlight the complementarity of using different models to functionally assesses splicing defects.

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“…In this case, retinal pigment epithelium derived from AMD-derived iPSCs was used to show impaired mitochondrial function under stress conditions and its link to the presence of the high-risk allele for the complement factor H ( CFH locus) ( 95 ). More generated iPSC lines from three patients carrying the rare variants in the CFH locus and suffering from AMD are available tools for further cellular studies and the development of novel treatments ( 96 ).…”

Section: Ipsc-based In Vitro Systems To Study Ieimentioning

confidence: 99%

In vitro systems to study inborn errors of immunity using human induced pluripotent stem cells

Nikolouli

Reichstein²,

Hansen³

et al. 2022

Front. Immunol.

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In the last two decades, the exponential progress in the field of genetics could reveal the genetic impact on the onset and progression of several diseases affecting the immune system. This knowledge has led to the discovery of more than 400 monogenic germline mutations, also known as “inborn errors of immunity (IEI)”. Given the rarity of various IEI and the clinical diversity as well as the limited available patients’ material, the continuous development of novel cell-based in vitro models to elucidate the cellular and molecular mechanisms involved in the pathogenesis of these diseases is imperative. Focusing on stem cell technologies, this review aims to provide an overview of the current available in vitro models used to study IEI and which could lay the foundation for new therapeutic approaches. We elaborate in particular on the use of induced pluripotent stem cell-based systems and their broad application in studying IEI by establishing also novel infection culture models. The review will critically discuss the current limitations or gaps in the field of stem cell technology as well as the future perspectives from the use of these cell culture systems.

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Generation and characterization of human induced pluripotent stem cells (iPSCs) from three patients with age-related macular degeneration carrying rare variants in the CFH gene

Cited by 4 publications

References 2 publications

Whole genome sequencing for USH2A-associated disease reveals several pathogenic deep-intronic variants that are amenable to splice correction

Whole genome sequencing for USH2A-associated disease reveals several pathogenic deep-intronic variants that are amenable to splice correction

Identification of a Complex Allele in IMPG2 as a Cause of Adult-Onset Vitelliform Macular Dystrophy

In vitro systems to study inborn errors of immunity using human induced pluripotent stem cells

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