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Identification and functional validation of a novel FBN1 variant in a Marfan syndrome family using a zebrafish model
Shitong Huang1,2, Jiansong Chen2, Qiuyu Wang3
1Department of Cardiac Surgical Intensive Care Unit, Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, 510080, China.
Background:
Marfan syndrome (MFS) is an inherited autosomal dominant disorder that affects connective tissue with an incidence of about 1 in 5,000 to 10,000 people. 90% of MFS is caused by mutations in the fibrillin-1 (FBN1) gene. We recruited a family with MFS phenotype in South China and identified a novel variant. This study investigated whether this genetic variant is pathogenic and the potential pathway related to lipid metabolism in MFS.
Methods:
A three-generation consanguineous family was recruited for this study. Whole exome sequencing (WES) was utilized on family members. The 3D structure of the protein was predicted using AlphaFold. CRISPR/Cas9 was applied to generate a similar fbn1 nonsense mutation (fbn1+/-) in zebrafish. RNA-seq analysis on zebrafish was performed to identify potential pathways related to MFS pathogenesis.
Results:
Our study identified a novel variant [NM_000138.5; c.7764 C > G: p.(Y2588*)] in FBN1 gene from the family and identified the same site mutation among the proband along with her son and daughter. Structural modeling showed the p.Y2588* mutation resulted from a truncated protein. Compared to wild-type zebrafish, the F2 generation fbn1+/- zebrafish exhibited MFS phenotype. RNA-seq analysis indicated that many genes related to leptin are up-regulating, which could affect bone development and adipose homeostasis.
Conclusion:
A novel variant was identified in FBN1 gene. In a zebrafish model, we found functional evidence supporting the pathogenicity of the detected nonsense mutation. Our research proposes a possible mechanism underlying the relationship between lipid metabolism and MFS. These findings can help improve the clinical diagnosis and treatment of MFS.
Insights
Researchers identified a new FBN1 gene variant causing Marfan syndrome (MFS). A zebrafish model confirmed the mutation
Area of Science:
- Genetics
- Molecular Biology
- Zebrafish Models
Background:
- Marfan syndrome (MFS) is a genetic connective tissue disorder affecting 1 in 5,000–10,000 individuals.
- Mutations in the fibrillin-1 (FBN1) gene cause 90% of MFS cases.
- A South China family with MFS phenotype was studied to identify a novel genetic variant.
Purpose of the Study:
- To investigate the pathogenicity of a novel FBN1 gene variant identified in a MFS family.
- To explore potential links between lipid metabolism and MFS pathogenesis.
Main Methods:
- Whole exome sequencing (WES) was performed on a three-generation consanguineous family.
- Protein 3D structure was predicted using AlphaFold.
- A novel nonsense mutation in FBN1 was created in zebrafish (fbn1+/-) using CRISPR/Cas9 for functional studies.
- RNA-sequencing (RNA-seq) was conducted on zebrafish to identify affected pathways.
Main Results:
- A novel pathogenic nonsense variant (c.7764 C>G: p.Y2588*) in the FBN1 gene was identified in the MFS family.
- Structural modeling predicted a truncated FBN1 protein due to the mutation.
- Zebrafish with the fbn1 nonsense mutation (fbn1+/-) displayed MFS phenotypes.
- RNA-seq revealed upregulation of leptin-related genes, suggesting an impact on bone development and adipose homeostasis.
Conclusions:
- A novel pathogenic FBN1 variant associated with Marfan syndrome was identified.
- Functional studies in zebrafish confirmed the pathogenicity of the nonsense mutation.
- The study suggests a potential mechanism linking lipid metabolism to MFS pathogenesis.
- Findings may aid in improving MFS clinical diagnosis and treatment.
