Tetralogy of Fallot: variants of MYH6 gene promoter and cellular functional analyses

• Published in: Pediatric research Vol. 96; no. 2; pp. 338 - 346
• Main Authors: Zuo, Ji-Yang, Chen, Huan-Xin, Yang, Qin, Liu, Zhi-Gang, He, Guo-Wei
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.07.2024; Nature Publishing Group
• Subjects: Animals; Basic Science Article; Binding sites; Cardiac Myosins - genetics; Cardiovascular disease; Cell Line; Child; Child, Preschool; Computational Biology; Congenital diseases; Family medical history; Female; Genes; Genetic Variation; Heart; HEK293 Cells; Hospitals; Humans; Male; Medical research; Medicine; Medicine & Public Health; Myosin Heavy Chains - genetics; Pediatric Surgery; Pediatrics; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Proteins; Surgery; Tetralogy of Fallot - genetics; Transcription factors; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 0031-3998; 1530-0447; 1530-0447
• DOI: 10.1038/s41390-023-02955-x

Background Tetralogy of Fallot (TOF) is a common form of congenital heart disease. The MYH6 gene has important effects on cardiovascular growth and development. Methods In 608 subjects, including 315 TOF patients, we investigated the MYH6 gene promoter variants and verified the effect on gene expression by using cellular functional experiments with three cell lines (HEK-293, HL-1, and H9C2 cells) and bioinformatics analysis. Results In the MYH6 gene promoter, 12 variants were identified from 608 subjects. Five variants were found only in patients with TOF and two of them (g.3384G>T and g.4518T>C) were novel. Electrophoretic mobility shift assay with three cell lines (HEK-293, HL-1, and H9C2) showed significant changes in the transcription factors bound by the promoter variants compared to the wild-type. Dual luciferase reporter showed that four of the five variants reduced the transcriptional activity of the MYH6 gene promoter ( p  < 0.05). Conclusions This study is the first to test the cellular function of variants in the promoter region of the MYH6 gene in patients with TOF, which provides new insights into the genetic basis of TOF and provides a basis for further study of the mechanism of TOF formation. Impact DNA from 608 human subjects was sequenced for MYH6 gene promoter region variants with five variants found only in TOF patients and two were novel. EMSA and dual luciferase reporter experiments in three cell lines found these variants pathological. Prediction by JASPAR database indicated that these variants alter the transcription factor binding sites. The study, for the first time, confirmed that there are variants at the MYH6 gene promoter region and these variants alter the cellular function. The variants found in this study suggest the possible pathological role in the formation of TOF.