Retinitis pigmentosa (RP) is a rare, progressive disease that affects photoreceptors and retinal pigment epithelial (RPE) cells with blindness as a final outcome. Despite high medical and social impact, there is currently no therapeutic options to slow down the progression of or cure the disease. The development of effective therapies was largely hindered by high genetic heterogeneity, inaccessible disease tissue, and unfaithful model organisms. The fact that components of ubiquitously expressed splicing factors lead to the retina-specific disease is an additional intriguing question. Herein, we sought to correlate the retinal cell-type-specific disease phenotype with the splicing profile shown by a patient with autosomal recessive RP, caused by a mutation in pre-mRNA splicing factor 8 (PRPF8). In order to get insight into the role of PRPF8 in homeostasis and disease, we capitalize on the ability to generate patient-specific RPE cells and reveal differentially expressed genes unique to RPE cells. We found that spliceosomal complex and ribosomal functions are crucial in determining cell-type specificity through differential expression and alternative splicing (AS) and that PRPF8 mutation causes global changes in splice site selection and exon inclusion that particularly affect genes involved in these cellular functions. This finding corroborates the hypothesis that retinal tissue identity is conferred by a specific splicing program and identifies retinal AS events as a framework toward the design of novel therapeutic opportunities.

• Klíčová slova
• PRPF8, RNA-Seq, RPE, alternative splicing, iPSC, pre-mRNA splicing, retinitis pigmentosa,
• Publikační typ
• časopisecké články MeSH

The human iPSC cell line, CARS-FiPS4F1 (ESi064-A), derived from dermal fibroblast from the apparently healthy carrier of the mutation of the gene SACSIN, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors. The pluripotency was assessed by immunocytochemistry and RT-PCR. This iPSC line can be used as control for Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) disease.

• MeSH
• dospělí MeSH
• indukované pluripotentní kmenové buňky metabolismus   MeSH
• Krüppel-like faktor 4 MeSH
• lidé MeSH
• mutace MeSH
• proteiny tepelného šoku genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• KLF4 protein, human MeSH Prohlížeč
• Krüppel-like faktor 4 MeSH
• proteiny tepelného šoku MeSH
• SACS protein, human MeSH Prohlížeč

The human iPSC cell line, ARS-FiPS4F1 (ESi063-A), derived from dermal fibroblast from the patient autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) caused by mutations on the gene SACSIN, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors. The pluripotency was assessed by immunocytochemistry and RT-PCR. Differentiation capacity was verified in vitro. This iPSC line can be further differentiated toward affected cells to better understand molecular mechanisms of disease and pathophysiology.

• MeSH
• buněčné linie MeSH
• indukované pluripotentní kmenové buňky metabolismus   MeSH
• Krüppel-like faktor 4 MeSH
• lidé MeSH
• mladiství MeSH
• mutace MeSH
• spinocerebelární ataxie vrozené   genetika   MeSH
• svalová spasticita genetika   MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH

The human iPSC cell line, GLC-FiPS4F1 (ESi047-A), derived from dermal fibroblast from the patient with congenital glaucoma caused by the mutation of the gene CYP1B1, was generated by non-integrative reprogramming technology using OCT3/4, SOX2, CMYC and KLF4 reprogramming factors.

• MeSH
• buněčná diferenciace MeSH
• buněčné kultury metody   MeSH
• buněčné linie MeSH
• cytochrom P450 CYP1B1 genetika   MeSH
• dospělí MeSH
• fibroblasty metabolismus   MeSH
• glaukom vrozené   genetika   MeSH
• indukované pluripotentní kmenové buňky cytologie   metabolismus   MeSH
• Krüppel-like faktor 4 MeSH
• lidé MeSH
• mutace genetika   MeSH
• Mycoplasma izolace a purifikace   MeSH
• přeprogramování buněk MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CYP1B1 protein, human MeSH Prohlížeč
• cytochrom P450 CYP1B1 MeSH
• KLF4 protein, human MeSH Prohlížeč
• Krüppel-like faktor 4 MeSH