Lack of CCDC146, a ubiquitous centriole and microtubule-associated protein, leads to non-syndromic male infertility in human and mouse

Rapport Preprint Lack of CCDC146, a ubiquitous centriole and microtubule-associated protein, leads to non-syndromic male infertility in human and mouse eng 2024-01-25 author Muroňová Jana author Kherraf Zine-Eddine author Giordani Elsa author Lambert Emeline author Eckert Simon author Cazin Caroline author Amiri-Yekta Amir author Court Magali author Chevalier Geneviève author Martinez Guillaume 0000-0002-7572-9096 846857 author Neirijnck Yasmine 834332 author Kuhne Françoise 23661 author Wehrli Lydia 834827 author Klena Nikolai 868290 author Hamel Virginie 0000-0001-5092-2343 846734 Département de biologie moléculaire et cellulaire (MOCEL) author De Macedo Lisa author Escoffier Breancon Jessica 0000-0001-8166-5845 815071 author Guichard Paul 0000-0002-0363-1049 845268 Département de biologie moléculaire et cellulaire (MOCEL) author Coutton Charles author Mustapha Selima Fourati Ben author Kharouf Mashmoud author Bouin Anne-Pascale author Zouari Raoudha author Thierry-Mieg Nicolas author Nef Serge 0000-0001-5462-0676 29062 Département de médecine génétique et développement (GEDEV) author Geimer Stefan author Loeuillet Corinne author Ray Pierre 0000-0003-1544-7449 author Arnoult Christophe 0000-0002-3753-5901 78 10.7554/eLife.86845.2 https://elifesciences.org/reviewed-preprints/86845v2 <p>Genetic mutations are a recurrent cause of male infertility. Multiple morphological abnormalities of the flagellum (MMAF) syndrome is a heterogeneous genetic disease, with which more than 50 genes have been linked. Nevertheless, for 50% of patients with this condition, no genetic cause is identified. From a study of a cohort of 167 MMAF patients, pathogenic bi-allelic mutations were identified in the CCDC146 gene in two patients. This gene encodes a poorly characterized centrosomal protein which we studied in detail here. First, protein localization was studied in two cell lines. We confirmed the centrosomal localization in somatic cells and showed that the protein also presents multiple microtubule-related localizations during mitotic division, suggesting that it is a microtubule-associated protein (MAP). To better understand the function of the protein at the sperm level, and the molecular pathogenesis of infertility associated with CCDC146 mutations, two genetically modified mouse models were created: a Ccdc146 knock-out (KO) and a knock-in (KI) expressing a HA-tagged CCDC146 protein. KO male mice were completely infertile, and sperm exhibited a phenotype identical to our two MMAF patient’s phenotype with CCDC146 mutations. No other pathology was observed, and the animals were viable. CCDC146 expression starts during late spermiogenesis, at the time of flagellum biogenesis. In the spermatozoon, the protein is conserved but is not localized to centrioles, unlike in somatic cells, rather it is present in the axoneme at the level of microtubule doublets. Expansion microscopy associated with the use of the detergent sarkosyl to solubilize microtubule doublets, suggest that the protein may be a microtubule inner protein (MIP). At the subcellular level, the absence of CCDC146 affected the formation, localization and morphology of all microtubule-based organelles such as the manchette, the head–tail coupling apparatus (HTCA), and the axoneme. Through this study, we have characterized a new genetic cause of infertility, identified a new factor in the formation and/or structure of the sperm axoneme, and demonstrated that the CCDC146 protein plays several cellular roles, depending on the cell type and the stages in the cell cycle.</p> Dewey 570 7daed2ee-1311-4824-9df9-2e51260a966a Département de biologie moléculaire et cellulaire 16484 false 8d549600-9ec0-4bf1-8218-f54ccb1cb4d2 Preprint Public Preprint-from-unpaywall.pdf 7486172 application/pdf CC BY-4.0 980f6ecdefaac54e877eb1c96bc1a6723b25f65d2473007a374a89bfc330cfe1 01829885-f1db-43fb-b11a-0565b1428853 Preprint Public 86845-v2.pdf 14866764 application/pdf CC BY d2c5cfc471bc8c5fef2340eb6cd9001f15abf4c69f0841112cc00439c8ce610c openAccess Faculté des sciences / Section de biologie / Département de biologie moléculaire et cellulaire

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Lack of CCDC146, a ubiquitous centriole and microtubule-associated protein, leads to non-syndromic male infertility in human and mouse				2024	105	124

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Martinez, Guillaume