BRCA1 相关蛋白 1 (BAP1) 中的致病性种系变异会导致 BAP1 肿瘤易感综合征 (BAP1-TPDS)。携带者尤其面临葡萄膜 (UM) 和皮肤黑色素瘤、恶性间皮瘤和透明细胞肾癌的风险。越来越多报道的 BAP1 变体中大约有一半缺乏准确的分类。正确解释致病性可以通过更好地了解 BAP1-TPDS 进行肿瘤筛查来改善患者的预后。我们使用 CRISPR-Cas9 在 HAP1 细胞中编辑了从 UM 患者中鉴定出的具有不同功能特征的 5 种罕见 BAP1 变体，并测定了它们对细胞粘附的影响/扩散（4 小时）和增殖（48 小时），使用 xCELLigence 实时分析系统以细胞指数 (CI) 进行测量。在 BAP1 敲除 HAP1 培养物中，细胞数量为 48 时野生型 (WT) 培养物的一半h (p = 0.00021)，较晚达到汇合，CI 降低了 78% (p < 0.0001)。 BAP1-TPDS 相关无效变体 c.67 1G>T 和 c.1780_1781insT，以及可能的致病性错义变体 c.281A>G 降低了粘附（所有 p ≤ 0.015）和增殖 74%-83%（所有 p ≤ 0.032） ）。另一种可能的致病性错义变异 c.680G>A 均减少了至少 50%（全部 p≤0.032），而用可能良性变异 c.1526C>T 编辑的细胞与 WT 类似地生长。BAP1 对于 HAP1 细胞的最佳适应性至关重要。致病性和可能致病性的 BAP1 变异降低了细胞适应性，这反映在粘附/扩散和增殖特性上。此外，中等影响是可以量化的。使用 CRISPR-Cas9 对 HAP1 进行变体建模，能够对内源表达系统中的编码区和非编码区变体进行功能分析。© 作者 2023。由牛津大学出版社出版。

Pathogenic germline variants in BRCA1-Associated Protein 1 (BAP1) cause BAP1 tumor predisposition syndrome (BAP1-TPDS). Carriers run especially a risk of uveal (UM) and cutaneous melanoma, malignant mesothelioma, and clear cell renal carcinoma. Approximately half of increasingly reported BAP1 variants lack accurate classification. Correct interpretation of pathogenicity can improve prognosis of the patients through tumor screening with better understanding of BAP1-TPDS.We edited five rare BAP1 variants with differing functional characteristics identified from patients with UM in HAP1 cells using CRISPR-Cas9 and assayed their effect on cell adhesion/spreading (at 4 h) and proliferation (at 48 h), measured as cell index (CI), using xCELLigence real-time analysis system.In BAP1 knockout HAP1 cultures, cell number was half of wild type (WT) cultures at 48 h (p = 0.00021), reaching confluence later, and CI was 78% reduced (p < 0.0001). BAP1-TPDS-associated null variants c.67+1G>T and c.1780_1781insT, and a likely pathogenic missense variant c.281A>G reduced adhesion (all p ≤ 0.015) and proliferation by 74%-83% (all p ≤ 0.032). Another likely pathogenic missense variant c.680G>A reduced both by at least 50% (all p ≤ 0.032), whereas cells edited with likely benign one c.1526C>T grew similarly to WT.BAP1 is essential for optimal fitness of HAP1 cells. Pathogenic and likely pathogenic BAP1 variants reduced cell fitness, reflected in adhesion/spreading and proliferation properties. Further, moderate effects were quantifiable. Variant modelling in HAP1 with CRISPR-Cas9 enabled functional analysis of coding and non-coding region variants in an endogenous expression system.© The Author(s) 2023. Published by Oxford University Press.