title: Localizing the RPGR protein along the cilium: a new method to determine efficacies to treat RPGR mutations.
abstract: Retinal dystrophies constitute a group of clinically and genetically heterogeneous diseases that cause visual impairment. As treatments are not readily available, readout assays performed in patient-derived cells can aid in the development and comparative analysis of therapeutic approaches. We describe a new method with which the localization of the retinitis pigmentosa GTPase regulator (RPGR) protein along the cilium can be used as a measure for treatment efficacy. In a patient-derived fibroblast cell line, we found that the RPGR protein is mislocalized along the ciliary axoneme. The patient carried a point mutation that leads to skipping of RPGR exon 10. We confirmed that this skipping is causative for the impaired localization of RPGR using a U7 small nuclear RNA (U7snRNA)-based antisense approach in control cells. Treatment of the patient-derived fibroblasts with therapeutic U1snRNA significantly corrected the proteins' mislocalization. In this proof of principle study, we show that detecting the RPGR protein along the cilium provides a reliable and quantifiable readout assay to evaluate the efficacy of therapies intended to correct or silence RPGR gene mutations. This method opens the possibility to compare different therapeutic agents, and thus facilitate the identification of treatment options for the clinically and molecularly complex RPGR-associated diseases.
authors: 1] Department of Human Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, Germany [2] Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland.
authors: R
authors: R
authors: Da Costa
authors: Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland.
authors: E
authors: E
authors: Glaus
authors: Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland.
authors: A
authors: A
authors: Tiwari
authors: 1] Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland [2] Department of Biology, ETH Zurich, Zurich, Switzerland.
authors: B
authors: B
authors: Kloeckener-Gruissem
authors: 1] Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland [2] Neuroscience Center Zurich, University of Zurich and ETH Zurich, Zurich, Switzerland [3] Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors: W
authors: W
authors: Berger
authors: 1] Department of Human Genetics, Faculty of Medicine and Health Sciences, University of Oldenburg, Oldenburg, Germany [2] Institute of Medical Molecular Genetics, University of Zurich, Schlieren, Switzerland [3] Research Center Neurosensory Science, Carl von Ossietzky Universität Oldenburg, Oldenburg, Germany.
authors: J
authors: J
authors: Neidhardt
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002923:Cilia / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005136:Eye Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017354:Point Mutation; D021381:Protein Transport; D012342:RNA, Small Nuclear / Q000235:genetics; D000067492:RNAi Therapeutics / Q000379:methods*; D058499:Retinal Dystrophies / Q000235:genetics* / Q000378:metabolism / Q000628:therapy
chemical_list: D005136:Eye Proteins; D012342:RNA, Small Nuclear; C099929:RPGR protein, human
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