Moldovan, Florina; Parent, Stefan; Barchi, Soraya; Hassan, Amani et Patten, Shunmoogum A. . Poc5 And Cilia Anomalies in Adolescent Idiopathic Scoliosis In: The Canadian Orthopaedic Association (COA) and Canadian Orthopaedic Research Society (CORS), 19–20 June 2020, virtuel.
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The etiology of adolescent idiopathic scoliosis (AIS) is largely unknown, but clinical observations revealed the role of hereditary and rapid growth in the development of this condition. More recently, several genes were suspected to cause or contribute to AIS. Our group identified gene variants of POC5 centriolar protein in a French and French-Canadian families with multiple members affected with AIS. We sought to expand on this study and to investigate for the role of POC5 gene and mutated protein. In this work, the potential pathogenic effect of mutated POC5 was investigated in vitro (human osteoblats cell culture) and in vivo in a zebrafish animal model.
To investigate the role of POC5 in AIS, we investigated subcellular localization of POC5 with respect to cilia in cells overexpressing wt or POC5 variants (C1286T, A429V) and in human osteoblasts from scoliotic patients carrying these POC5 variants and normal control cells (in vitro study). We also created a loss-of-function model in zebrafish (in vivo study). The role of POC5 was investigated by: 1) mass spectroscopy analysis and co-immunoprecipitation to identify differences in binding partners between the wild-type (wt POC5 and mut POC5 protein; 2) immunolocalization of POC5 wt and mut proteins at the cellular level; 3) histology and immunohistochemistry performed on tissues from wt (control) and scoliotic (poc5 mut) zebrafish.
Our work identified several interacting partners with POC5, and documented functional connections with respect to cilia and centrosome dysfunction. A number of ciliary proteins were identified to be interacting with wt POC5 but not mut POC5. At the cellular level, localization and co-localisation of wt POC5 and mut POC5 protein with alpha acetylated tubulin (cilia marker), confirmed the consequence of the mutation on subcellular location with respect to cilium structure, length and staining intensity of cilia. In vivo, several defects in the retina were identified in mut poc5 zebrafish compared wt zebrafish. Finally, using different markers for retinal layers and acetylated tubulin, the defects were localized in ganglion cell layer and cones of the retina.
Our findings confirm the involvement of POC5 in scoliosis. A role of POC5 with respect to the primary cilia was attributed. These findings open new avenues for the understanding the primary causes of AIS at the molecular and physiological levels.
| Type de document: | Document issu d'une conférence ou d'un atelier |
|---|---|
| Informations complémentaires: | Orthopaedic Proceedings Vol. 103-B, No. SUPP_3 p.42 https://online.boneandjoint.org.uk/toc/procs/103-B/SUPP_3 |
| Mots-clés libres: | - |
| Centre: | Centre INRS-Institut Armand Frappier |
| Date de dépôt: | 14 juill. 2021 15:43 |
| Dernière modification: | 14 juill. 2021 15:43 |
| URI: | https://espace.inrs.ca/id/eprint/11863 |
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