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 manifestation 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 high bleeding tendency, as well as an abnormal expansion of progenitor cells. The gp9SMU15 zebrafish can be used as a BSS animal model as GP9 roles 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 increasing 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 potential BSS therapeutic strategy development.

中文翻译：

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斑马鱼Bernard-Soulier综合征模型的建立。

血小板在血栓形成和止血中起重要作用。异常止血可导致自发性或严重的创伤后出血。Bernard-Soulier 综合征 (BSS) 是一种罕见的遗传性出血性疾病，由 GPIb-IX-V 复合体的完全定量缺陷引起。GP9的多重突变导致BSS的临床表现。了解GP9在血小板生成中的作用和潜在机制，建立合适的BSS动物模型，对于了解疾病发病机制和改善其医疗管理具有重要意义。在这里，通过使用 CRISPR-Cas9 技术，我们创建了一个斑马鱼 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高出血倾向，以及祖细胞的异常扩增。gp9SMU15 斑马鱼可用作 BSS 动物模型，因为 GP9 在血小板生成中的作用从斑马鱼到哺乳动物都是高度保守的。利用 BSS 模型，我们通过体内功能测定验证了临床 GP9 突变，并测试了增加血小板的临床药物。因此，遗传的 BSS 斑马鱼模型可能有助于体内验证患者衍生的具有不确定意义的 GP9 变体和潜在的 BSS 治疗策略开发。