Emerging evidence suggests that children conceived through assisted reproductive technology (ART) have a higher risk of congenital heart defects (CHDs) even when there is no family history. De novo mutation (DNM) is a well-known cause of sporadic congenital diseases; however, whether ART procedures increase the number of germline DNM (gDNM) has not yet been well studied. Here, we performed whole-genome sequencing of 1137 individuals from 160 families conceived through ART and 205 families conceived spontaneously. Children conceived via ART carried 4.59 more gDNMs than children conceived spontaneously, including 3.32 paternal and 1.26 maternal DNMs, after correcting for parental age at conception, cigarette smoking, alcohol drinking, and exercise behaviors. Paternal DNMs in offspring conceived via ART are characterized by C>T substitutions at CpG sites, which potentially affect protein-coding genes and are significantly associated with the increased risk of CHD. In addition, the accumulation of non-coding functional mutations was independently associated with CHD and 87.9% of the mutations were originated from the father. Among ART offspring, infertility of the father was associated with elevated paternal DNMs; usage of both recombinant and urinary follicle-stimulating hormone and high-dosage human chorionic gonadotropin trigger was associated with an increase of maternal DNMs. In sum, the increased gDNMs in offspring conceived by ART were primarily originated from fathers, indicating that ART itself may not be a major reason for the accumulation of gDNMs. Our findings emphasize the importance of evaluating the germline status of the fathers in families with the use of ART.

新出现的证据表明，即使没有家族史，通过辅助生殖技术 (ART) 受孕的儿童也有较高的先天性心脏缺陷 (CHD) 风险。从头突变（DNM）是散发性先天性疾病的众所周知的原因。然而，ART 程序是否会增加生殖系 DNM (gDNM) 的数量尚未得到很好的研究。在这里，我们对来自 160 个通过 ART 受孕的家庭和 205 个自发受孕的家庭的 1137 名个体进行了全基因组测序。在校正了父母的受孕年龄、吸烟、饮酒和运动行为后，通过 ART 受孕的孩子比自然受孕的孩子携带的 gDNM 多 4.59 个，其中包括 3.32 个父亲和 1.26 个母亲 DNM。通过 ART 受孕的后代中的父系 DNM 的特征是 C> CpG 位点的 T 取代可能影响蛋白质编码基因，并与 CHD 风险增加显着相关。此外，非编码功能突变的积累与冠心病独立相关，87.9%的突变来源于父亲。在 ART 后代中，父亲不育与父亲 DNM 升高有关；重组和尿促卵泡激素以及高剂量人绒毛膜促性腺激素触发器的使用与母体 DNM 的增加有关。综上所述，ART 孕育的后代中 gDNMs 增加主要来自父亲，这表明 ART 本身可能不是 gDNMs 积累的主要原因。