Validation of GWAS-derived novel regulators of musculoskeletal homeostasis

Aim & Background: Validation of the role of novel musculoskeletal genes discovered by GWAS is required to make a bridge between in-silico discoveries and application for improving human life quality. Advanced genome editing technologies such as CRISPR-Cas9 allows prioritizing and further validating novel musculoskeletal regulators in an animal model. The life cycle of zebrafish and high genetic similarity to human make it an efficient model for fast-time genetics manipulations. Most importantly, genetic determinants that control bone formation are highly conserved between zebrafish and mammals. The aim of our study is to validate novel genes, which popped out from human GWAS data, as associated with musculoskeletal homeostasis using the zebrafish model. Methods: We performed preliminary expression screening analysis for GWAS candidates. Based on expression assay we selected relevant candidate genes for generating stable mutant lines. The expression screening was done by RNA in-situ hybridization in zebrafish intact and 4 dpa regenerated (4 days post-amputation) fins and by qPCR in bone tissues (scales, operculum) and muscles tissues (skeletal muscles, heart).Results&Conclusions: The adamtsl3, fubp3, jag1a, mef2ca, wnt4a, wls genes pointed out the involvement in bone homeostasis, during bone regeneration in zebrafish fins. The high expression in muscles compared to bone was observed for fam210aa, fam210ab, mef2ca. Novel candidate genes expressed in muscle tissue are also relevant for further research. Therefore, we conclude to focus on adamtsl3, fam210aa, fam210ab, fubp3 as relevant candidates for generating mutant lines, as further research is required to identify the role of these novel genes in bone homeostasis.