室间隔缺损与HAND2基因罕见变异的关联分析

李美琨; 逄淑超; 闫波

doi:10.7499/j.issn.1008-8830.2212057

PDF(883 KB)

HTML

中国当代儿科杂志 ›› 2023, Vol. 25 ›› Issue (4) : 388-393. DOI: 10.7499/j.issn.1008-8830.2212057

论著·临床研究

室间隔缺损与HAND2基因罕见变异的关联分析

李美琨¹, 逄淑超^2,3, 闫波^1,4

作者信息 +

Association of ventricular septal defect with rare variations of the HAND2 gene

LI Mei-Kun, PANG Shu-Chao, YAN Bo

Author information +

文章历史 +

摘要

目的对室间隔缺损（ventricular septal defect，VSD）与HAND2基因启动子区罕见变异的关联性进行分析，并对相关分子机制进行初步探索。方法收集349例VSD组患儿和345例健康对照组儿童血液样本，通过聚合酶链反应扩增目的片段并进行测序，确定HAND2基因启动子区的罕见变异位点。使用双荧光素酶检测进行变异位点功能分析，并通过电泳迁移率变动分析进行分子机制研究，同时使用TRANSFAC和JASPAR数据库预测转录因子。结果通过测序发现3个变异位点（g.173530852A>G、g.173531173A>G和g.173531213C>G）只出现在10例VSD患儿的HAND2基因启动子上，其中4例患儿仅有1个变异位点。双荧光素酶检测显示g.173531213C>G降低了HAND2基因启动子转录活性。电泳迁移率变动分析和转录因子预测提示g.173531213C>G产生了1个转录因子结合位点。结论 HAND2基因启动子区的罕见变异g.173531213C>G可能通过影响转录因子的结合，进而参与VSD的发生发展。［中国当代儿科杂志，2023，25（4）：388-393］

Abstract

Objective To study the association of ventricular septal defect (VSD) with rare variations in the promoter region of HAND2 gene, as well as related molecular mechanisms. Methods Blood samples were collected from 349 children with VSD and 345 healthy controls. The target fragments were amplified by polymerase chain reaction and sequenced to identify the rare variation sites in the promoter region of the HAND2 gene. Dual-luciferase reporter assay was used to perform a functional analysis of the variation sites. Electrophoretic mobility shift assay (EMSA) was used to investigate related molecular mechanisms. TRANSFAC and JASPAR databases were used to predict transcription factors. Results Sequencing revealed that three variation sites (g.173530852A>G, g.173531173A>G, and g.173531213C>G) were only observed in the promoter region of the HAND2 gene in 10 children with VSD, among whom 4 children had only one variation site. The dual-luciferase reporter assay revealed that g.173531213C>G reduced the transcriptional activity of the HAND2 gene promoter. EMSA and transcription factor prediction revealed that g.173531213C>G created a binding site for transcription factor. Conclusions The rare variation, g.173531213C>G, in the promoter region of the HAND2 gene participates in the development and progression of VSD possibly by affecting the binding of transcription factors. Citation:Chinese Journal of Contemporary Pediatrics, 2023, 25(4): 388-393

导出引用

李美琨, 逄淑超, 闫波. 室间隔缺损与HAND2基因罕见变异的关联分析[J]. 中国当代儿科杂志. 2023, 25(4): 388-393 https://doi.org/10.7499/j.issn.1008-8830.2212057

LI Mei-Kun, PANG Shu-Chao, YAN Bo. Association of ventricular septal defect with rare variations of the HAND2 gene[J]. Chinese Journal of Contemporary Pediatrics. 2023, 25(4): 388-393 https://doi.org/10.7499/j.issn.1008-8830.2212057

参考文献

1 Tsao CW, Aday AW, Almarzooq ZI, et al. Heart disease and stroke statistics—2022 update: a report from the American Heart Association[J]. Circulation, 2022, 145(8): e153-e639. PMID: 35078371. DOI: 10.1161/CIR.0000000000001052.
2 Muto M, Sugita K, Matsuba T, et al. How should we treat representative neonatal surgical diseases with congenital heart disease?[J]. Pediatr Surg Int, 2022, 38(9): 1235-1240. PMID: 35838788. DOI: 10.1007/s00383-022-05178-z.
3 Luo M, Wang T, Huang P, et al. Association of maternal betaine-homocysteine methyltransferase (BHMT) and BHMT2 genes polymorphisms with congenital heart disease in offspring[J]. Reprod Sci, 2023, 30(1): 309-325. PMID: 35835902. DOI: 10.1007/s43032-022-01029-3.
4 George RM, Firulli AB. Hand factors in cardiac development[J]. Anat Rec (Hoboken), 2019, 302(1): 101-107. PMID: 30288953. PMCID: PMC6312500. DOI: 10.1002/ar.23910.
5 Morton SU, Quiat D, Seidman JG, et al. Genomic frontiers in congenital heart disease[J]. Nat Rev Cardiol, 2022, 19(1): 26-42. PMID: 34272501. PMCID: PMC9236191. DOI: 10.1038/s41569-021-00587-4.
6 Bruneau BG. Signaling and transcriptional networks in heart development and regeneration[J]. Cold Spring Harb Perspect Biol, 2013, 5(3): a008292. PMID: 23457256. PMCID: PMC3578359. DOI: 10.1101/cshperspect.a008292.
7 Meilhac SM, Buckingham ME. The deployment of cell lineages that form the mammalian heart[J]. Nat Rev Cardiol, 2018, 15(11): 705-724. PMID: 30266935. DOI: 10.1038/s41569-018-0086-9.
8 Zhao K, Yang Z. The second heart field: the first 20 years[J]. Mamm Genome, 2022. Epub ahead of print. PMID: 36550326. DOI: 10.1007/s00335-022-09975-8.
9 Moore-Morris T, van Vliet PP, Andelfinger G, et al. Role of epigenetics in cardiac development and congenital diseases[J]. Physiol Rev, 2018, 98(4): 2453-2475. PMID: 30156497. DOI: 10.1152/physrev.00048.2017.
10 Vincentz JW, Toolan KP, Zhang W, et al. Hand factor ablation causes defective left ventricular chamber development and compromised adult cardiac function[J]. PLoS Genet, 2017, 13(7): e1006922. PMID: 28732025. PMCID: PMC5544250. DOI: 10.1371/journal.pgen.1006922.
11 Firulli AB, McFadden DG, Lin Q, et al. Heart and extra-embryonic mesodermal defects in mouse embryos lacking the bHLH transcription factor Hand1[J]. Nat Genet, 1998, 18(3): 266-270. PMID: 9500550. DOI: 10.1038/ng0398-266.
12 Srivastava D, Thomas T, Lin Q, et al. Regulation of cardiac mesodermal and neural crest development by the bHLH transcription factor, dHAND[J]. Nat Genet, 1997, 16(2): 154-160. PMID: 9171826. DOI: 10.1038/ng0697-154.
13 Ritter O, Haase H, Schulte HD, et al. Remodeling of the hypertrophied human myocardium by cardiac bHLH transcription factors[J]. J Cell Biochem, 1999, 74(4): 551-561. PMID: 10440925. DOI: 10.1002/(sici)1097-4644(19990915)74:4<551::aid-jcb5>3.3.co;2-0.
14 Tsuchihashi T, Maeda J, Shin CH, et al. Hand2 function in second heart field progenitors is essential for cardiogenesis[J]. Dev Biol, 2011, 351(1): 62-69. PMID: 21185281. PMCID: PMC3039109. DOI: 10.1016/j.ydbio.2010.12.023.
15 Shen L, Li XF, Shen AD, et al. Transcription factor HAND2 mutations in sporadic Chinese patients with congenital heart disease[J]. Chin Med J (Engl), 2010, 123(13): 1623-1627. PMID: 20819618.
16 Sun YM, Wang J, Qiu XB, et al. A HAND2 loss-of-function mutation causes familial ventricular septal defect and pulmonary stenosis[J]. G3 (Bethesda), 2016, 6(4): 987-992. PMID: 26865696. PMCID: PMC4825666. DOI: 10.1534/g3.115.026518.
17 Vincentz JW, Firulli BA, Toolan KP, et al. HAND transcription factors cooperatively specify the aorta and pulmonary trunk[J]. Dev Biol, 2021, 476: 1-10. PMID: 33757801. PMCID: PMC8172455. DOI: 10.1016/j.ydbio.2021.03.011.
18 Kuchenbaecker K, Appel EVR. Assessing rare variation in complex traits[J]. Methods Mol Biol, 2018, 1793: 51-71. PMID: 29876891. DOI: 10.1007/978-1-4939-7868-7_5.
19 Auer PL, Lettre G. Rare variant association studies: considerations, challenges and opportunities[J]. Genome Med, 2015, 7(1): 16. PMID: 25709717. PMCID: PMC4337325. DOI: 10.1186/s13073-015-0138-2.
20 Chen W, Coombes BJ, Larson NB. Recent advances and challenges of rare variant association analysis in the biobank sequencing era[J]. Front Genet, 2022, 13: 1014947. PMID: 36276986. PMCID: PMC9582646. DOI: 10.3389/fgene.2022.1014947.
21 Moorman AF, Christoffels VM. Cardiac chamber formation: development, genes, and evolution[J]. Physiol Rev, 2003, 83(4): 1223-1267. PMID: 14506305. DOI: 10.1152/physrev.00006.2003.
22 Harris IS, Black BL. Development of the endocardium[J]. Pediatr Cardiol, 2010, 31(3): 391-399. PMID: 20135106. PMCID: PMC2836465. DOI: 10.1007/s00246-010-9642-8.
23 Poelmann RE, Gittenberger-de Groot AC. Development and evolution of the metazoan heart[J]. Dev Dyn, 2019, 248(8): 634-656. PMID: 31063648. PMCID: PMC6767493. DOI: 10.1002/dvdy.45.
24 Tanaka AJ, Cho MT, Willaert R, et al. De novo variants in EBF3 are associated with hypotonia, developmental delay, intellectual disability, and autism[J]. Cold Spring Harb Mol Case Stud, 2017, 3(6): a002097. PMID: 29162653. PMCID: PMC5701309. DOI: 10.1101/mcs.a002097.
25 Yang L, Lin M, Ruan WJ, et al. Nkx2-1: a novel tumor biomarker of lung cancer[J]. J Zhejiang Univ Sci B, 2012, 13(11): 855-866. PMID: 23125078. PMCID: PMC3494024. DOI: 10.1631/jzus.B1100382.
26 Nameki N, Takizawa M, Suzuki T, et al. Structural basis for the interaction between the first SURP domain of the SF3A1 subunit in U2 snRNP and the human splicing factor SF1[J]. Protein Sci, 2022, 31(10): e4437. PMID: 36173164. PMCID: PMC9514218. DOI: 10.1002/pro.4437.