The Role of TBX20 Gene Mutations in the Pathogenesis of Congenital Heart Disease: Functional Analysis and Genetic Association Study

Objective: This study aims to investigate the pathogenic role of TBX20 gene mutations in congenital heart disease (CHD), evaluate their impact on cardiac development and function, and quantify the association between TBX20 gene mutations and CHD risk through meta-analysis. Methods: Genetic screening of 353 CHD patients and 350 healthy children was conducted using high-throughput sequencing technology to identify TBX20 gene mutations. Homology modeling and molecular dynamics simulations were employed to assess the mutations' effects on the structure and function of the TBX20 protein. The impact of these mutations on the cardiac cell phenotype was further verified through in vitro experiments. A meta-analysis, incorporating literature search and quality assessment, was conducted to quantitatively evaluate the relationship between TBX20 gene mutations and CHD risk. Results: Two critical mutations in the TBX20 gene (missense mutation I121F and synonymous mutation T262T) were identified, and bioinformatics predictions along with molecular modeling revealed potential decreases in protein structural stability. The meta-analysis, including five studies, indicated that TBX20 gene mutations significantly increase CHD risk (pooled OR=5.73, 95%CI=[2.54, 12.91]). The influence of mutant TBX20 on its mRNA expression levels and downstream target gene ANF promoter activity further supported this finding. Sensitivity analysis and publication bias assessment confirmed the robustness of the results. Conclusion: This study confirms that TBX20 gene mutations play a significant role in the pathogenesis of CHD, affecting protein structure and function and significantly increasing CHD risk. These findings offer new insights into the genetic basis of CHD and may impact future diagnostic and therapeutic strategies.