Early disruption of photoreceptor cell architecture and loss of vision in a humanized pig model of usher syndromes
2022; Springer Nature; Volume: 14; Issue: 4 Linguagem: Inglês
10.15252/emmm.202114817
ISSN1757-4684
AutoresSophia Grotz, Jessica Schäfer, Kirsten A. Wunderlich, Zdeňka Ellederová, Hannah Auch, Andrea Bähr, Petra Vochozková, Janet Fadl, Vanessa Arnold, Taras Ardan, Miroslav Veith, Gianluca Santamaria, G. Dhom, Wolfgang Hitzl, Barbara Keßler, C. Eckardt, Joshua P. Klein, Anna Brýmová, Joshua Linnert, Mayuko Kurome, Valeri Zakharchenko, Andrea Fischer, Andreas Blutke, Anna Döring, Stepanka Suchankova, Jiří Popelář, Eduardo Rodríguez‐Bocanegra, Julia Dlugaiczyk, Hans Straka, Helen May‐Simera, Weiwei Wang, Karl‐Ludwig Laugwitz, Luk H. Vandenberghe, Eckhard Wolf, Kerstin Nagel‐Wolfrum, Tobias Peters, Jan Motlík, M. Dominik Fischer, Uwe Wolfrum, Nikolai Klymiuk,
Tópico(s)CRISPR and Genetic Engineering
ResumoArticle7 March 2022Open Access Source DataTransparent process Early disruption of photoreceptor cell architecture and loss of vision in a humanized pig model of usher syndromes Sophia Grotz Sophia Grotz Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Jessica Schäfer Jessica Schäfer Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Kirsten A Wunderlich Corresponding Author Kirsten A Wunderlich [email protected] Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation, Methodology Search for more papers by this author Zdenka Ellederova Zdenka Ellederova Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Methodology, Project administration Search for more papers by this author Hannah Auch Hannah Auch Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Validation, Investigation, Methodology Search for more papers by this author Andrea Bähr Andrea Bähr Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology Search for more papers by this author Petra Runa-Vochozkova Petra Runa-Vochozkova orcid.org/0000-0003-3691-0340 Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology Search for more papers by this author Janet Fadl Janet Fadl Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Vanessa Arnold Vanessa Arnold Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Taras Ardan Taras Ardan Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Miroslav Veith Miroslav Veith orcid.org/0000-0001-6254-436X Ophthalmology Clinic, University Hospital Kralovske Vinohrady, Praha, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Gianluca Santamaria Gianluca Santamaria Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Software, Formal analysis, Methodology Search for more papers by this author Georg Dhom Georg Dhom Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Wolfgang Hitzl Wolfgang Hitzl orcid.org/0000-0002-7696-1479 Biostatistics and Data Science, Paracelsus Medical University, Salzburg, Austria Contribution: Formal analysis, Methodology Search for more papers by this author Barbara Kessler Barbara Kessler orcid.org/0000-0001-7036-238X Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Methodology Search for more papers by this author Christian Eckardt Christian Eckardt Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Joshua Klein Joshua Klein Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Anna Brymova Anna Brymova Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Investigation Search for more papers by this author Joshua Linnert Joshua Linnert Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Mayuko Kurome Mayuko Kurome Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Methodology Search for more papers by this author Valeri Zakharchenko Valeri Zakharchenko Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Search for more papers by this author Andrea Fischer Andrea Fischer Veterinary Faculty, Small Animal Clinics, LMU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology, Writing - review & editing Search for more papers by this author Andreas Blutke Andreas Blutke Institute of Experimental Genetics, Helmholtz Center Munich, Neuherberg, Germany Contribution: Investigation, Methodology, Writing - review & editing Search for more papers by this author Anna Döring Anna Döring Veterinary Faculty, Small Animal Clinics, LMU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Stepanka Suchankova Stepanka Suchankova Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Jiri Popelar Jiri Popelar Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic Contribution: Supervision Search for more papers by this author Eduardo Rodríguez-Bocanegra Eduardo Rodríguez-Bocanegra Centre for Ophthalmology, University Eye Hospital, University Hospital Tübingen, Tübingen, Germany Institute for Ophthalmic Research, Centre for Ophthalmology, University Hospital Tübingen, Tübingen, Germany Contribution: Validation Search for more papers by this author Julia Dlugaiczyk Julia Dlugaiczyk Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Zurich (USZ), University of Zurich, Zurich, Switzerland Contribution: Supervision Search for more papers by this author Hans Straka Hans Straka Faculty of Biology, LMU Munich, Planegg, Germany Contribution: Formal analysis Search for more papers by this author Helen May-Simera Helen May-Simera orcid.org/0000-0002-9477-4036 Institute of Molecular Physiology, Cilia Biology, JGU Mainz, Mainz, Germany Contribution: Conceptualization, Investigation, Methodology, Writing - review & editing Search for more papers by this author Weiwei Wang Weiwei Wang orcid.org/0000-0003-3518-9328 Grousbeck Gene Therapy Center, Mass Eye and Ear and Harvard Medical School, Boston, MA, USA Contribution: Resources Search for more papers by this author Karl-Ludwig Laugwitz Karl-Ludwig Laugwitz Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Conceptualization, Methodology, Writing - review & editing Search for more papers by this author Luk H Vandenberghe Luk H Vandenberghe Grousbeck Gene Therapy Center, Mass Eye and Ear and Harvard Medical School, Boston, MA, USA Contribution: Resources Search for more papers by this author Eckhard Wolf Eckhard Wolf orcid.org/0000-0002-0430-9510 Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Conceptualization, Methodology, Writing - review & editing Search for more papers by this author Kerstin Nagel-Wolfrum Kerstin Nagel-Wolfrum Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Institute of Developmental Biology and Neurobiology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Conceptualization, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Tobias Peters Tobias Peters orcid.org/0000-0002-9187-4904 Centre for Ophthalmology, University Eye Hospital, University Hospital Tübingen, Tübingen, Germany Institute for Ophthalmic Research, Centre for Ophthalmology, University Hospital Tübingen, Tübingen, Germany Contribution: Formal analysis Search for more papers by this author Jan Motlik Jan Motlik Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Supervision, Methodology, Project administration, Writing - review & editing Search for more papers by this author M Dominik Fischer M Dominik Fischer Oxford Eye Hospital, Oxford University NHS Foundation Trust, Oxford, UK Nuffield Laboratory of Ophthalmology, NDCN, University of Oxford, Oxford, UK Search for more papers by this author Uwe Wolfrum Uwe Wolfrum orcid.org/0000-0002-4756-5872 Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Conceptualization, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Nikolai Klymiuk Corresponding Author Nikolai Klymiuk [email protected] orcid.org/0000-0003-3532-1659 Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Conceptualization, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Project administration, Writing - review & editing Search for more papers by this author Sophia Grotz Sophia Grotz Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Jessica Schäfer Jessica Schäfer Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Kirsten A Wunderlich Corresponding Author Kirsten A Wunderlich [email protected] Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation, Methodology Search for more papers by this author Zdenka Ellederova Zdenka Ellederova Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Methodology, Project administration Search for more papers by this author Hannah Auch Hannah Auch Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Validation, Investigation, Methodology Search for more papers by this author Andrea Bähr Andrea Bähr Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology Search for more papers by this author Petra Runa-Vochozkova Petra Runa-Vochozkova orcid.org/0000-0003-3691-0340 Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology Search for more papers by this author Janet Fadl Janet Fadl Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Vanessa Arnold Vanessa Arnold Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Taras Ardan Taras Ardan Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Miroslav Veith Miroslav Veith orcid.org/0000-0001-6254-436X Ophthalmology Clinic, University Hospital Kralovske Vinohrady, Praha, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Gianluca Santamaria Gianluca Santamaria Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Software, Formal analysis, Methodology Search for more papers by this author Georg Dhom Georg Dhom Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Wolfgang Hitzl Wolfgang Hitzl orcid.org/0000-0002-7696-1479 Biostatistics and Data Science, Paracelsus Medical University, Salzburg, Austria Contribution: Formal analysis, Methodology Search for more papers by this author Barbara Kessler Barbara Kessler orcid.org/0000-0001-7036-238X Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Methodology Search for more papers by this author Christian Eckardt Christian Eckardt Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Joshua Klein Joshua Klein Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Anna Brymova Anna Brymova Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Investigation Search for more papers by this author Joshua Linnert Joshua Linnert Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Investigation Search for more papers by this author Mayuko Kurome Mayuko Kurome Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Methodology Search for more papers by this author Valeri Zakharchenko Valeri Zakharchenko Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Search for more papers by this author Andrea Fischer Andrea Fischer Veterinary Faculty, Small Animal Clinics, LMU Munich, Munich, Germany Contribution: Supervision, Investigation, Methodology, Writing - review & editing Search for more papers by this author Andreas Blutke Andreas Blutke Institute of Experimental Genetics, Helmholtz Center Munich, Neuherberg, Germany Contribution: Investigation, Methodology, Writing - review & editing Search for more papers by this author Anna Döring Anna Döring Veterinary Faculty, Small Animal Clinics, LMU Munich, Munich, Germany Contribution: Investigation Search for more papers by this author Stepanka Suchankova Stepanka Suchankova Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic Contribution: Investigation, Methodology Search for more papers by this author Jiri Popelar Jiri Popelar Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic Contribution: Supervision Search for more papers by this author Eduardo Rodríguez-Bocanegra Eduardo Rodríguez-Bocanegra Centre for Ophthalmology, University Eye Hospital, University Hospital Tübingen, Tübingen, Germany Institute for Ophthalmic Research, Centre for Ophthalmology, University Hospital Tübingen, Tübingen, Germany Contribution: Validation Search for more papers by this author Julia Dlugaiczyk Julia Dlugaiczyk Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Zurich (USZ), University of Zurich, Zurich, Switzerland Contribution: Supervision Search for more papers by this author Hans Straka Hans Straka Faculty of Biology, LMU Munich, Planegg, Germany Contribution: Formal analysis Search for more papers by this author Helen May-Simera Helen May-Simera orcid.org/0000-0002-9477-4036 Institute of Molecular Physiology, Cilia Biology, JGU Mainz, Mainz, Germany Contribution: Conceptualization, Investigation, Methodology, Writing - review & editing Search for more papers by this author Weiwei Wang Weiwei Wang orcid.org/0000-0003-3518-9328 Grousbeck Gene Therapy Center, Mass Eye and Ear and Harvard Medical School, Boston, MA, USA Contribution: Resources Search for more papers by this author Karl-Ludwig Laugwitz Karl-Ludwig Laugwitz Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Conceptualization, Methodology, Writing - review & editing Search for more papers by this author Luk H Vandenberghe Luk H Vandenberghe Grousbeck Gene Therapy Center, Mass Eye and Ear and Harvard Medical School, Boston, MA, USA Contribution: Resources Search for more papers by this author Eckhard Wolf Eckhard Wolf orcid.org/0000-0002-0430-9510 Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany Center for Innovative Medical Models, LMU Munich, Munich, Germany Contribution: Conceptualization, Methodology, Writing - review & editing Search for more papers by this author Kerstin Nagel-Wolfrum Kerstin Nagel-Wolfrum Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Institute of Developmental Biology and Neurobiology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Conceptualization, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Tobias Peters Tobias Peters orcid.org/0000-0002-9187-4904 Centre for Ophthalmology, University Eye Hospital, University Hospital Tübingen, Tübingen, Germany Institute for Ophthalmic Research, Centre for Ophthalmology, University Hospital Tübingen, Tübingen, Germany Contribution: Formal analysis Search for more papers by this author Jan Motlik Jan Motlik Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic Contribution: Supervision, Methodology, Project administration, Writing - review & editing Search for more papers by this author M Dominik Fischer M Dominik Fischer Oxford Eye Hospital, Oxford University NHS Foundation Trust, Oxford, UK Nuffield Laboratory of Ophthalmology, NDCN, University of Oxford, Oxford, UK Search for more papers by this author Uwe Wolfrum Uwe Wolfrum orcid.org/0000-0002-4756-5872 Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany Contribution: Conceptualization, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Writing - review & editing Search for more papers by this author Nikolai Klymiuk Corresponding Author Nikolai Klymiuk [email protected] orcid.org/0000-0003-3532-1659 Center for Innovative Medical Models, LMU Munich, Munich, Germany Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany Contribution: Conceptualization, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Writing - original draft, Project administration, Writing - review & editing Search for more papers by this author Author Information Sophia Grotz1,2,†, Jessica Schäfer3,†, Kirsten A Wunderlich *,3,20, Zdenka Ellederova4, Hannah Auch1,2, Andrea Bähr2,5, Petra Runa-Vochozkova5, Janet Fadl1,3, Vanessa Arnold3, Taras Ardan4, Miroslav Veith6, Gianluca Santamaria5, Georg Dhom1,2, Wolfgang Hitzl7, Barbara Kessler1,2, Christian Eckardt2,5, Joshua Klein3, Anna Brymova4, Joshua Linnert3, Mayuko Kurome1,2, Valeri Zakharchenko1,2, Andrea Fischer8, Andreas Blutke9, Anna Döring8, Stepanka Suchankova10, Jiri Popelar10, Eduardo Rodríguez-Bocanegra11,12, Julia Dlugaiczyk13, Hans Straka14, Helen May-Simera15, Weiwei Wang16, Karl-Ludwig Laugwitz5, Luk H Vandenberghe16, Eckhard Wolf1,2, Kerstin Nagel-Wolfrum3,17, Tobias Peters11,12, Jan Motlik4, M Dominik Fischer18,19, Uwe Wolfrum3,‡ and Nikolai Klymiuk *,2,5,‡ 1Chair of Molecular Animal Breeding and Biotechnology, LMU Munich, Munich, Germany 2Center for Innovative Medical Models, LMU Munich, Munich, Germany 3Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University (JGU), Mainz, Germany 4Institute of Animal Physiology and Genetics, Czech Academy of Science, Libechov, Czech Republic 5Large Animal Models in Cardiovascular Research, Internal Medical Department I, TU Munich, Munich, Germany 6Ophthalmology Clinic, University Hospital Kralovske Vinohrady, Praha, Czech Republic 7Biostatistics and Data Science, Paracelsus Medical University, Salzburg, Austria 8Veterinary Faculty, Small Animal Clinics, LMU Munich, Munich, Germany 9Institute of Experimental Genetics, Helmholtz Center Munich, Neuherberg, Germany 10Institute of Experimental Medicine, Czech Academy of Science, Prague, Czech Republic 11Centre for Ophthalmology, University Eye Hospital, University Hospital Tübingen, Tübingen, Germany 12Institute for Ophthalmic Research, Centre for Ophthalmology, University Hospital Tübingen, Tübingen, Germany 13Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Zurich (USZ), University of Zurich, Zurich, Switzerland 14Faculty of Biology, LMU Munich, Planegg, Germany 15Institute of Molecular Physiology, Cilia Biology, JGU Mainz, Mainz, Germany 16Grousbeck Gene Therapy Center, Mass Eye and Ear and Harvard Medical School, Boston, MA, USA 17Institute of Developmental Biology and Neurobiology, Johannes Gutenberg University (JGU), Mainz, Germany 18Oxford Eye Hospital, Oxford University NHS Foundation Trust, Oxford, UK 19Nuffield Laboratory of Ophthalmology, NDCN, University of Oxford, Oxford, UK 20Present address: Department of Physiological Genomics, LMU Munich, Munich, Germany † These authors contributed equally to this work as first authors ‡ These authors contributed equally to this work as senior authors *Corresponding author. Tel: +49 6131 3925148; E-mail: [email protected] *Corresponding author. Tel: +49 151 522 95 921; E-mail: [email protected] EMBO Mol Med (2022)14:e14817https://doi.org/10.15252/emmm.202114817 PDFDownload PDF of article text and main figures. Peer ReviewDownload a summary of the editorial decision process including editorial decision letters, reviewer comments and author responses to feedback. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Usher syndrome (USH) is the most common form of monogenic deaf-blindness. Loss of vision is untreatable and there are no suitable animal models for testing therapeutic strategies of the ocular constituent of USH, so far. By introducing a human mutation into the harmonin-encoding USH1C gene in pigs, we generated the first translational animal model for USH type 1 with characteristic hearing defect, vestibular dysfunction, and visual impairment. Changes in photoreceptor architecture, quantitative motion analysis, and electroretinography were characteristics of the reduced retinal virtue in USH1C pigs. Fibroblasts from USH1C pigs or USH1C patients showed significantly elongated primary cilia, confirming USH as a true and general ciliopathy. Primary cells also proved their capacity for assessing the therapeutic potential of CRISPR/Cas-mediated gene repair or gene therapy in vitro. AAV-based delivery of harmonin into the eye of USH1C pigs indicated therapeutic efficacy in vivo. Synopsis The full phenotypic spectrum of Usher Syndrome is reflected in a pig model carrying a patient-specific mutation after partial humanization of the porcine USH1C gene. Retinal function in USH1C pigs was improved when harmonin expression was reconstituted by AAV-mediated gene therapy. Partial humanization of the porcine USH1C gene was facilitated by the high degree of sequence conservation in the N-terminal region of harmonin. Vision loss within the first year of life in USH1C pigs was consistently indicated by behavior tests, clinical examination and morphological analysis. Early onset of vision loss was correlated to disrupted photoreceptor cell architecture. Ciliopathy mechanisms and their therapeutic correction were investigated in primary cells of USH1C individuals. Local application and therapeutic efficacy of AAV-mediated treatments was examined in USH1C pigs in vivo. The paper explained Problem Due to the lack of sufficient animal models, the mechanisms of vision loss in Usher syndrome (USH) are incompletely understood and therapies cannot be sufficiently tested. As a consequence, patients inevitably suffer blindness. Results A pig model for USH reflects the combination of deafness, vestibular dysfunction, and vision loss seen in USH patients. Visual impairment in USH1C pigs, as documented by ophthalmologic examinations and parameterized behavioral tests, is accompanied by morphologic changes in photoreceptor cells. Further, photoreceptor cilia and primary cilia of fibroblasts were consistently elongated in USH1C pigs and facilitated the testing of gene supplementation and gene correction as potential therapeutics. Exploratory gene therapy experiments prove the amelioration of visual function in USH1C pigs. Impact The USH1C pig is the first animal model for studying the ocular component of the Usher syndrome in vivo, at the cellular and the molecular level. Our achievements will promote future research in two relevant ways: (i) reversible changes in primary cilia length make fibroblasts a simple, cheap and robust in vitro evaluation tool for testing treatment options in USH. (ii) The consistent multi-disciplinary examination of the ocular phenotype paves the way for a comprehensive evaluation of vision in pigs and therefore towards a more reliable judgment of pre-clinical therapy studies in the USH1C pig model in vivo. Introduction Usher syndrome (USH) is the most common form of inherited deaf-blindness in humans (Mathur & Yang, 2015). USH is clinically and genetically heterogeneous, with at least 10 genes assigned to three clinical USH types (Moller et al, 1989; Toms et al, 2020). The most severe of them is USH1, characterized by profound hearing loss from birth on, vestibular areflexia, and pre-pubertal onset of retinitis pigmentosa (RP). In patients, the congenital sensorineural hearing impairment can be compensated with cochlear implants, but no therapeutic option is presently available for the ocular disease constituent. Thus, most USH cases lead to severe visual impairment and blindness over time, adding a substantial psychological component to the clinical symptoms (Lonborg-Moller et al, 2020). Development of therapies and understanding of USH pathogenesis in the retina is arduous as existing rodent models for USH reflect the human deficits in the inner ear, but show only a very mild, if any, retinal phenotype (Lentz et al, 2007; Williams, 2008; El-Amraoui & Petit, 2014). This has been attributed to substantial differences between mouse and human in anatomy and cellular composition of the retina and stimulated discussion about alternative model species. Pigs reflect structure and function of the human eye much better than mice as they have a similar size, a comparable rod/cone ratio and a cone-rich region, the so-called visual streak (Chandler et al, 1999; Hendrickson & Hicks, 2002). Specifically, calyceal processes (CP), cellular extravaginations at the transition of the inner segment (IS) to the outer segment (OS) of photoreceptor cells (PRC) appear in many species, including human and pig, but not rodents (Wolfrum et al, 2010; Sahly et al, 2012; El-Amraoui & Petit, 2014). Various pig models have been described for retinal degeneration, with the majority of them induced by compounds or light (Li et al, 2009; Wang et al, 2011; Monés et al, 2016) or additive transgenesis (Petters et al, 1997; Sommer et al, 2011; Ross et al, 2012; Kostic et al, 2013). As the latter do not truly reflect the genetic constellation in patients, the phenotype was, however, difficult to interpret and limited the utility of these models. We therefore hypothesized that introducing a patient-specific mutation into a partially humanized USH1C gene would facilitate a better genotype–phenotype correlation of an USH1C pig model. USH1C comprises 28 exons with alternative splicing giving rise to numerous tra
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