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Heidi Barbara Stoehr, Marlene Schwarzfischer, Lisa-Marie Braun, Alexander Aslanidis, Thomas Langmann, Ulrich Kellner; Investigating the pathomechanism of FAM161A-associated retinal disease in primary ciliated cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3192.
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© ARVO (1962-2015); The Authors (2016-present)
Loss-of-function mutations in the FAM161A gene cause autosomal recessive retinitis pigmentosa (RP28). We sought to obtain primary ciliated cells lacking FAM161A as a model system to study the pathomechanism underlying FAM161A-associated ciliopathies.
DNA testing was performed via targeted gene panel diagnostics in a patient with simplex retinitis pigmentosa. Clinical assessment included visual acuity, visual fields, fundus (FAF) and near-infrared autofluorescence (NIA) and spectral domain optical coherence tomography (SD-OCT). RT-PCR was used for mRNA expression analysis. Human dermal fibroblasts and mouse lung fibroblasts were immunostained with antibodies against FAM161A, FAM161B and markers specific for microtubules and the Golgi apparatus.
A 31-year old female from Spain was found to carry a novel homozygous c.730delA;p.Met244* stop mutation in exon 3 of the FAM161A gene. Clinical examination revealed a visual acuity of 20/100 on both eyes, markedly constricted visual fields, FAF and NIA findings characteristic for retinitis pigmentosa. The SD-OCT showed preserved photoreceptor layers at the posterior pole without macular edema. RT-PCR analysis revealed endogenous mRNA expression of FAM161A and FAM161B, a direct interaction partner of FAM161A, in control fibroblast cultures. FAM161A co-localized with acetylated α-tubulin in primary cilia and FAM161B overlapped with γ-tubulin in the basal body. Neither FAM161A nor FAM161B showed overlapping localization with the Golgi marker GM-130.
FAM161A is present in primary cilia of fibroblast cultures. Therefore, fibroblasts or other ciliated cells such as urine-derived renal epithelial cells cultivated from patients or a Fam161a-deficient mouse model can be used for further ex vivo analysis of molecular mechanisms underlying FAM161A-associated disease.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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