CCDC78: Unveiling the Function of a Novel Gene Associated with Hereditary Myopathy
Abstract
:1. Introduction
2. Materials and Methods
2.1. Standard Protocol Approvals and Patient Consents
2.2. Morphological Analysis of Muscle Biopsies and HeLa Cells
2.3. DNA Analysis
2.4. Cells Culture
2.5. RNA Extraction
2.6. Transcripts Analysis
2.7. Transcriptome Profiling
2.8. RT-qPCR
2.9. Western Blotting Study
2.10. Co-Immunoprecipitation (Co-Ip) Assay
2.11. Mass Spectroscopy Study
2.12. nLC-nESI-HRMS/MS
2.13. Filtering and Selecting Interactors Candidates for Validation
2.14. Three-Dimensional (3D) Modelling
3. Results
3.1. The Nonsense CCDC78 Mutation Leads to a Dystrophic Muscle Process Mainly Involving the SR
3.2. Human Tissues Express Multiple CCDC78 Isoforms, and the Nonsense CCDC78 Variant Leads to a Mutant Allele That Is Mostly Degraded by NMD
3.3. The Dysfunction of CCDC78 Leads to an Upregulation of Genes Involved in the SR
3.4. CCDC78 Interacts with Two Important SR Proteins: SERCA1 and CASQ1
3.5. CCDC78 Interacts with Other Proteins outside the SR: MYH1, ACTN2, and ACTA1
3.6. CCDC78 Does Not Directly Interact with RyR1
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Lopergolo, D.; Gallus, G.N.; Pieraccini, G.; Boscaro, F.; Berti, G.; Serni, G.; Volpi, N.; Formichi, P.; Bianchi, S.; Cassandrini, D.; et al. CCDC78: Unveiling the Function of a Novel Gene Associated with Hereditary Myopathy. Cells 2024, 13, 1504. https://doi.org/10.3390/cells13171504
Lopergolo D, Gallus GN, Pieraccini G, Boscaro F, Berti G, Serni G, Volpi N, Formichi P, Bianchi S, Cassandrini D, et al. CCDC78: Unveiling the Function of a Novel Gene Associated with Hereditary Myopathy. Cells. 2024; 13(17):1504. https://doi.org/10.3390/cells13171504
Chicago/Turabian StyleLopergolo, Diego, Gian Nicola Gallus, Giuseppe Pieraccini, Francesca Boscaro, Gianna Berti, Giovanni Serni, Nila Volpi, Patrizia Formichi, Silvia Bianchi, Denise Cassandrini, and et al. 2024. "CCDC78: Unveiling the Function of a Novel Gene Associated with Hereditary Myopathy" Cells 13, no. 17: 1504. https://doi.org/10.3390/cells13171504