c0cf72700daf9c3057e5f819f000a990 pmc_10968961.pdf 6e9ec572590a8b4a5ae92fd3bdd56b62d852881e pmc_10968961.pdf 5ef1dc9ee361e62329442f9214869c597954728c484e55031333d0c96309acbc pmc_10968961.pdf Title: Primary Ciliary Dyskinesia and Retinitis Pigmentosa: Novel RPGR Variant and Possible Modifier Gene Subject: We report a novel RPGR missense variant co-segregated with a familial X-linked retinitis pigmentosa (XLRP) case. The brothers were hemizygous for this variant, but only the proband presented with primary ciliary dyskinesia (PCD). Thus, we aimed to elucidate the role of the RPGR variant and other modifier genes in the phenotypic variability observed in the family and its impact on motile cilia. The pathogenicity of the variant on the RPGR protein was evaluated by in vitro studies transiently transfecting the mutated RPGR gene, and immunofluorescence analysis on nasal brushing samples. Whole-exome sequencing was conducted to identify potential modifier variants. In vitro studies showed that the mutated RPGR protein could not localise to the cilium and impaired cilium formation. Accordingly, RPGR was abnormally distributed in the siblings’ nasal brushing samples. In addition, a missense variant in CEP290 was identified. The concurrent RPGR variant influenced ciliary mislocalisation of the protein. We provide a comprehensive characterisation of motile cilia in this XLRP family, with only the proband presenting PCD symptoms. The variant’s pathogenicity was confirmed, although it alone does not explain the respiratory symptoms. Finally, the CEP290 gene may be a potential modifier for respiratory symptoms in patients with RPGR mutations.12pt Keywords: primary cilia; motile cilia; primary ciliary dyskinesia; retinitis pigmentosa; PCD; XLRP; RPGR; CEP290; modifier gene Author: Noelia Baz-Redón, Laura Sánchez-Bellver, Mónica Fernández-Cancio, Sandra Rovira-Amigo, Thomas Burgoyne, Rai Ranjit, Virginia Aquino, Noemí Toro-Barrios, Rosario Carmona, Eva Polverino, Maria Cols, Antonio Moreno-Galdó, Núria Camats-Tarruella and Gemma Marfany Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Sat Mar 16 10:12:40 2024 CET ModDate: Sat Mar 16 10:25:40 2024 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 11155023 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JCYIDT+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 AMYURO+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 EUBVIG+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 UQJEXV+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 NKIZAJ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 32 0 DSBBYX+VnURWPalladioL Type 1 Custom yes yes yes 37 0 PFKOHU+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 53 0 MDJLZU+EURM10 Type 1 Builtin yes yes yes 73 0 FKHIHB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 88 0 FKHIHD+PalatinoLinotype TrueType WinAnsi yes yes no 91 0 FKHIHF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 94 0 FXXUVH+CMSY10 Type 1 Builtin yes yes yes 104 0 FKHIHB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 117 0 FKHIHD+PalatinoLinotype TrueType WinAnsi yes yes no 120 0 FKHIHF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 123 0 FKHIHB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 142 0 FKHIHC+PalatinoLinotype CID TrueType Identity-H yes yes yes 145 0 FKHIHD+PalatinoLinotype TrueType WinAnsi yes yes no 151 0 FKHIHF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 154 0 FKHIHB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 173 0 FKHIHC+PalatinoLinotype CID TrueType Identity-H yes yes yes 176 0 FKHIHD+PalatinoLinotype TrueType WinAnsi yes yes no 182 0 FKHIHF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 185 0 FKHIHB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 202 0 FKHIHC+PalatinoLinotype CID TrueType Identity-H yes yes yes 205 0 FKHIHD+PalatinoLinotype TrueType WinAnsi yes yes no 211 0 FKHIHF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 214 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-06-18 02:41:17 CEST RepresentationInformation: pmc_10968961.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-06-12 02:48:45 CEST Size: 11155023 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 483 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Patients Destination: subsection.2.1 Item: Title: Primary Ciliary Dyskinesia Diagnostic Evaluation Destination: subsection.2.2 Item: Title: In Silico Studies of the Candidate Variant Destination: subsection.2.3 Item: Title: In Vitro Functional Studies Destination: subsection.2.4 Item: Title: Immunofluorescence Analyses of RPGR and CEP290 in Nasal Brushing Samples Destination: subsection.2.5 Item: Title: Statistics Destination: subsection.2.6 Item: Title: Results Destination: section.3 Children: Item: Title: PCD Clinical Manifestation and Diagnosis Destination: subsection.3.1 Item: Title: Identification of the Pathogenic RPGR Variant Destination: subsection.3.2 Item: Title: RPGR Immunofluorescence Analysis in Respiratory Ciliated Cells Destination: subsection.3.3 Item: Title: Additive Effect of Modifier Genes on the PCD Phenotype Destination: subsection.3.4 Item: Title: CEP290 Immunofluorescence Analysis in Respiratory Ciliated Cells Destination: subsection.3.5 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Primary Ciliary Dyskinesia and Retinitis Pigmentosa: Novel RPGR Variant and Possible Modifier Gene Author: Noelia Baz-Redón, Laura Sánchez-Bellver, Mónica Fernández-Cancio, Sandra Rovira-Amigo, Thomas Burgoyne, Rai Ranjit, Virginia Aquino, Noemí Toro-Barrios, Rosario Carmona, Eva Polverino, Maria Cols, Antonio Moreno-Galdó, Núria Camats-Tarruella and Gemma Marfany Subject: We report a novel RPGR missense variant co-segregated with a familial X-linked retinitis pigmentosa (XLRP) case. The brothers were hemizygous for this variant, but only the proband presented with primary ciliary dyskinesia (PCD). Thus, we aimed to elucidate the role of the RPGR variant and other modifier genes in the phenotypic variability observed in the family and its impact on motile cilia. The pathogenicity of the variant on the RPGR protein was evaluated by in vitro studies transiently transfecting the mutated RPGR gene, and immunofluorescence analysis on nasal brushing samples. Whole-exome sequencing was conducted to identify potential modifier variants. In vitro studies showed that the mutated RPGR protein could not localise to the cilium and impaired cilium formation. Accordingly, RPGR was abnormally distributed in the siblings’ nasal brushing samples. In addition, a missense variant in CEP290 was identified. The concurrent RPGR variant influenced ciliary mislocalisation of the protein. We provide a comprehensive characterisation of motile cilia in this XLRP family, with only the proband presenting PCD symptoms. The variant’s pathogenicity was confirmed, although it alone does not explain the respiratory symptoms. 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