Abstract：Objective To explore the differences of NKX2.5 and TBX5 gene mutations between in vitro fertilization (IVF) children with congenital heart disease (CHD) and naturally conceived children with CHD. Methods Blood samples from 68 IVF children with CHD and 98 naturally conceived children with CHD were collected. The mutations in coding regions 1 and 2 of the NKX2.5 gene, and coding regions 4, 5, and 8 of the TBX5 gene were examined by polymerase chain reaction (PCR) and DNA sequencing. Results An A-to-G mutation at nucleotide 63 (c.63A > G) in coding region 1 of the NKX2.5 gene was found in both IVF and naturally conceived children with CHD. There were no significant differences in genotype and allele frequencies at c.63A > G locus of the NKX2.5 gene between the two groups. No mutations were detected in coding region 2 of the NKX2.5 gene and coding regions 4, 5 and 8 of the TBX5 gene. Conclusions There is no difference in NKX2.5 and TBX5 gene mutations between IVF and naturally conceived children with CHD. Therefore, it is presumed that assisted reproductive technology may not lead to mutations in the NKX2.5 and TBX5 genes.
YANG Jing-Hui,XU Xiao-Yan,MI Hong-Ying et al. NKX2.5 and TBX5 gene mutations in in vitro fertilization children with congenital heart disease[J]. CJCP, 2017, 19(6): 652-657.
Schott JJ, Benson DW, Basson CT, et al. Congenital heart disease caused by mutations in the transcription factor NKX2-5[J]. Science, 1998, 281(5373):108-111.
[5]
McElhinney DB, Geiger E, Blinder J, et al. NKX2.5 mutations in patients with congenital heart disease[J]. J Am Coll Cardiol, 2003, 42(9):1650-1655.
[6]
Reamon-Buettner SM, Hecker H, Spanel-Borowski K, et al. Novel NKX2-5 mutations in diseased heart tissues of patients with cardiac malformations[J]. Am J Pathol, 2004, 164(6):2117-2125.
[7]
Bruneau BG, Nemer G, Schmitt JP, et al. A murine model of Holt-Oram syndrome defines roles of the T-Box transcription factor Tbx5 in cardiogenesis and disease[J]. Cell, 2001, 106(6):709-721.
[8]
Reamon-Buettner SM, Borlak J. TBX5 mutations in non-Holt-Oram syndrome (HOS) malformed hearts[J]. Hum Mutat, 2004, 24(1):104.
[9]
Zhang WM, Li XF, Ma ZY, et al. GATA4 and NKX2.5 gene analysis in Chinese Uygur patients with congenital heart disease[J]. Chin Med J, 2009, 122(4):416-419.
[10]
Pabst S, Wollnik B, Rohmann E, et al. A novel stop mutation truncating critical regions of the cardiac transcription factor NKX2-5 in a large family with autosomal-dominant inherited congenital heart disease[J]. Clin Res Cardiol, 2008, 97(1):39-42.
[11]
Dott M, Rasmussen SA, Hogue CJ, et al. Association between pregnancy intention and reproductive-health related behaviors before and after pregnancy recognition, National Birth Defects Prevention Study, 1997-2002[J]. Matern Child Health J, 2010, 14(3):373-381.
[12]
Koivurova S, Hartikainen AL, Gissler M, et al. Neonatal outcome and congenital malformations in children born after in-vitro fertilization[J]. Hum Reprod, 2002, 17(5):1391-1398.
[13]
Hoffman JI, Kaplan S. The incidence of congenital heart disease[J]. J Am Coll Cardiol, 2002, 39(12):1890-1900.
[14]
He A, Kong SW, Ma Q, et al. Co-occupancy by multiple cardiac transcription factors identifies transcriptional enhancers active in heart[J]. Proc Natl Acad Sci, 2011, 108(14):5632-5637.
[15]
Clark CD, Zhang B, Lee B, et al. Evolutionary conservation of Nkx2.5 autoregulation in the second heart field[J]. Dev Biol, 2013, 374(1):198-209.
[16]
Greulich F, Rudat C, Kispert A. Mechanisms of T-box gene function in the developing heart[J]. Cardiovasc Res, 2011, 91(2):212-222.
[17]
Gioli-Pereira L, Pereira AC, Mesquita SM, et al. NKX2.5 mutations in patients with non-syndromic congenital heart disease[J]. Int J Cardiol, 2009, 138(3):261-265.
[18]
Stallmeyer B, Fenge H, Nowak-Göttl U, et al. Mutational spectrum in the cardiac transcription factor gene NKX2.5(CSX) associated with congenital heart disease[J]. Clin Genet, 2010, 78(6):533-540.
[19]
Reamon-Buettner SM, Borlak J. NKX2-5:an update on this hypermutable homeodomain protein and its role in human congenital heart disease (CHD)[J]. Hum Mutat, 2010, 31(11):1185-1194.
[20]
Hatemi AC, Güleç C, Cine N, et al. Sequence variations of NKX2-5 and HAND1 genes in patients with atrial isomerism[J]. Anadolu Kardiyol Derg, 2011, 11(4):319-328.
Wang Z, Zou L, Zhong R, et al. Associations between two genetic variants in NKX2-5 and risk of congenital heart disease in Chinese population:a meta-analysis[J]. PLoS One, 2013, 8(8):e70979.
Tararbit K, Lelong N, Thieulin AC, et al. The risk for four specific congenital heart defects associated with assisted reproductive techniques:a population-based evaluation[J]. Hum Reprod, 2013, 28(2):367-374.
[30]
Bahtiyar MO, Campbell K, Dulay AT, et al. Is the rate of congenital heart defects detected by fetal echocardiography among pregnancies conceived by in vitro fertilization really increased?:a case-historical control study[J]. J Ultrasound Med, 2010, 29(6):917-922.
