高危神经母细胞瘤是一种复杂的遗传性疾病，尽管进行了强化的多模式治疗，但仍在50%以上的患者中致命。通过基因组范围的关联研究 (GWAS) 和下一代测序 (NGS)，我们发现 BARD1 中普通的单核苷酸多态性 (SNP) 和罕见的致病性 (P) 或可能致病性 (LP) 生殖细胞系功能缺陷 (LOF) 变异在神经母细胞瘤患者中富集。这些发现的功能意义尚不清楚。我们将 BARD1 基因型与正常组织和神经母细胞瘤中的表达相关联，以及肿瘤中的 DNA 损伤负担。为了验证生殖细胞病变 P-LP BARD1 变异的功能后果，我们使用 CRISPR/Cas9 生成了携带杂合 BARD1 LOF 变异 (R112*，R150*，E287fs 和 Q564*) 的神经母细胞瘤 (IMR-5) 和对照 (RPE1) 细胞模型，并通过 NGS 和功能分析测量 DNA 修复效率来量化这些细胞的基因组不稳定性。普通和罕见的神经母细胞瘤相关 BARD1 生殖细胞病变显著与 BARD1 mRNA 水平降低和 DNA 损伤负担增加相关。利用杂合 BARD1 LOF 变异细胞模型，我们在功能上验证了这种关联与 DNA 修复效率低下相关。BARD1 LOF 变异同源细胞表现出对 Cas9 诱导的 DNA 损伤修复效率降低，DNA 双链断裂位点上 RAD51 聚焦形成无效，以及在体内外对顺铂和多聚腺苷酸核糖聚合酶 (PARP) 抑制剂的敏感性增强。综上所述，我们证明了生殖细胞 BARD1 变异破坏了 DNA 修复的准确性。这是神经母细胞瘤发生的一个基本分子机制，可能具有重要的治疗意义。© 版权所有 2023 作者. 由牛津大学出版社出版。保留所有权利。如需授权，请发送电子邮件至：journals.permissions@oup.com。

High-risk neuroblastoma is a complex genetic disease that is lethal in over 50% of patients despite intense multimodal therapy. Through genome-wide association studies (GWAS) and next-generation sequencing (NGS), we have identified common single nucleotide polymorphisms (SNPs) and rare, pathogenic (P) or likely pathogenic (LP) germline loss-of-function (LOF) variants in BARD1 enriched in neuroblastoma patients. The functional implications of these findings remain poorly understood.We correlated BARD1 genotype with expression in normal tissues and neuroblastomas, along with the burden of DNA damage in tumors. To validate the functional consequences of germline P-LP BARD1 variants, we used CRISPR/Cas9 to generate isogenic neuroblastoma (IMR-5) and control (RPE1) cellular models harboring heterozygous BARD1 LOF variants (R112*, R150*, E287fs, and Q564*) and quantified genomic instability in these cells via NGS and with functional assays measuring the efficiency of DNA repair.Both common and rare neuroblastoma associated BARD1 germline variants were significantly associated with lower levels of BARD1 mRNA and an increased burden of DNA damage. Using isogenic heterozygous BARD1 LOF variant cellular models, we functionally validated this association with inefficient DNA repair. BARD1 LOF variant isogenic cells exhibited reduced efficiency in repairing Cas9-induced DNA damage, ineffective RAD51 focus formation at DNA double-strand break sites, and enhanced sensitivity to cisplatin and poly-ADP ribose polymerase (PARP) inhibition both in vitro and in vivo.Taken together, we demonstrate that germline BARD1 variants disrupt DNA repair fidelity. This is a fundamental molecular mechanism contributing to neuroblastoma initiation that may have important therapeutic implications.© The Author(s) 2023. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.