Key Laboratory of Zebrafish Modeling and Drug Screening for Human Diseases of Guangdong Higher Education Institutes, Department of Developmental Biology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China; Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology, Guangzhou 510006.
Division of Cell, Developmental and Integrative Biology, School of Medicine, South China University of Technology, Guangzhou 510006.
Haematologica. 2022 Jul 1;107(7):1655-1668. doi: 10.3324/haematol.2021.278893.
Platelets play an essential role in thrombosis and hemostasis. Abnormal hemostasis can cause spontaneous or severe post-traumatic bleeding. Bernard-Soulier syndrome (BSS) is a rare inherited bleeding disorder caused by a complete quantitative deficiency in the GPIb-IX-V complex. Multiple mutations in GP9 lead to the clinical manifestations of BSS. Understanding the roles and underlying mechanisms of GP9 in thrombopoiesis and establishing a proper animal model of BSS would be valuable to understand the disease pathogenesis and to improve its medical management. Here, by using CRISPR-Cas9 technology, we created a zebrafish gp9SMU15 mutant to model human BSS. Disruption of zebrafish gp9 led to thrombocytopenia and a pronounced bleeding tendency, as well as an abnormal expansion of progenitor cells. The gp9SMU15 zebrafish can be used as a BSS animal model as the roles of GP9 in thrombocytopoiesis are highly conserved from zebrafish to mammals. Utilizing the BSS model, we verified the clinical GP9 mutations by in vivo functional assay and tested clinical drugs for their ability to increase platelets. Thus, the inherited BSS zebrafish model could be of benefit for in vivo verification of patient-derived GP9 variants of uncertain significance and for the development of potential therapeutic strategies for BSS.
血小板在血栓形成和止血中起着至关重要的作用。异常止血会导致自发性或严重创伤后出血。伯纳德-苏利耶综合征（BSS）是一种罕见的遗传性出血性疾病，由 GPIb-IX-V 复合物完全定量缺乏引起。GP9 的多种突变导致 BSS 的临床表现。了解 GP9 在血小板生成中的作用和潜在机制，并建立适当的 BSS 动物模型，对于了解疾病发病机制和改善其医疗管理将是有价值的。在这里，我们使用 CRISPR-Cas9 技术创建了一种斑马鱼 gp9SMU15 突变体来模拟人类 BSS。斑马鱼 gp9 的破坏导致血小板减少和明显的出血倾向，以及前体细胞的异常扩张。gp9SMU15 斑马鱼可用作 BSS 动物模型，因为 GP9 在从斑马鱼到哺乳动物的血小板生成中的作用高度保守。利用 BSS 模型，我们通过体内功能测定验证了临床 GP9 突变，并测试了临床药物增加血小板的能力。因此，遗传性 BSS 斑马鱼模型可能有助于对不确定意义的患者衍生 GP9 变异体进行体内验证，并为 BSS 的潜在治疗策略的开发提供帮助。
