PMC10014717_01
Patient's clinical characteristics. (A) Bilateral elbow wrist joint examination, demonstrating a partial fusion of bones at the proximal end of both the ulna and radius, widening of the radial ulnar joint at the distal end, and poor alignment. (B) Electrocardiography before (top) and after surgery (bottom). (C) Computed tomography angiography of the cardiac structure before (top) and after surgery (bottom). (D) Resting and adenosine load-gated myocardial perfusion imaging demonstrated that there was no typical reversible abnormal perfusion in the left ventricular myocardium, and the left ventricular cardiac function index was normal. (E) Three-dimensionally printed heart model used to determine the surgical approach.
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A bilateral elbow wrist joint examination demonstrated the partial fusion of the bones at the proximal end of both the ulna and radius, widening of the radial ulnar joint at the distal end, and poor alignment, suggesting congenital dysplasia in the wrist and elbow joints (Figure 1A). Outpatient coronary computed tomography angiography and angiography after admission revealed that her left coronary artery originated from the right coronary sinus and the slender left coronary artery was contorted between the aorta and the pulmonary artery (Figures 1B, C). Resting and adenosine load-gated myocardial perfusion imaging revealed no typical reversible abnormal perfusion in the left ventricular myocardium and showed that the left ventricular cardiac function index was normal (Figure 1D). A three-dimensional printed model was used to determine the appropriate surgical approach (Figure 1E).
Effects of TBX5 expression on cardiomyocytes. (A) TBX5 expression after TBX5 knockdown in the HL-1 cell line by transfection of small interfering RNAs. (B) Expression of TEKT2, TEKT4, and SPTB decreased after TBX5 knockdown, indicating that they are downstream of TBX5. (C) Chromatin immunoprecipitation analysis showed that binding of TEKT2, TEKT4, and SPTB DNA fragments to TBX5 decreased after TBX5 knockdown, demonstrating that TBX5 promoted TEKT2, TEKT4, and SPTB transcription by binding to the promoters. beta-Actin was used as the internal control. (D) Motif of binding sites between TBX5 protein and the promoters of the DNA fragments. (E) Sequence alignment between TBX5 3'-UTRs, including rs883079 and rs10850326, and microRNAs (miRNA), including hsa-miR-98-5p, hsa-miR-10a, hsa-miR-10b, and hsa-miR-182-5p. The bases on the rs883079 and rs10850326 are labeled yellow. (F) Venn diagram of single-nucleotide polymorphisms and mutations in the HOS patient's family determined using whole-exome sequencing. *p < 0.05, **p < 0.01, and ***p < 0.001; ns, not significant.
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We explored the effects of TBX5 on the HL-1 cell line with TBX5 knockdown via transfection of small interfering RNAs (Ubigene, Guangzhou, China; Figure 2A). Quantitative polymerase chain reaction analysis demonstrated that the expression of TEKT2, TEKT4, and SPTB decreased after TBX5 knockdown (Figure 2B), and chromatin immunoprecipitation analysis further revealed that TBX5 binds to TEKT2, TEKT4, and SPTB promoter regions to promote gene transcription (Figures 2C, D; Table 2). TBX5 3'-UTR that included rs883079, while only hsa-miR-10a and hsa-miR-182-5p could bind to the region that included rs10850326, demonstrating that the increased binding of miRNAs and TBX5 3'-UTR in HOS further decreased TBX5 expression (Figure 2E). These SNPs and familial mutations may be associated with the differential expression of TBX5 (Figure 2F).