Gene Correction of Wiskott-Aldrich syndrome iPS Cells Rescues Proplatelet Defects and Improves Platelet Size.

IF 5 2区 医学 Q1 HEMATOLOGY
Praewphan Ingrungruanglert, Sarinya Phodang, Pramuk Amarinthnukrowh, Phattarawan Meehart, Pornpitra Pratedrat, Narissara Suratannon, Vorasuk Shotelersuk, Kanya Suphapeetiporn, Nipan Israsena
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Abstract

Wiskott-Aldrich syndrome (WAS) is a severe X-linked disorder caused by loss-of-function mutations in the WAS gene, responsible for encoding WAS protein (WASP), a key regulator of the actin cytoskeleton in all hematopoietic cells, except red blood cells. The mechanism underlying microthrombocytopenia, a distinctive feature of WAS and a major contributor to mortality, remains not fully elucidated. In this study, using different gene-editing strategies, we corrected mutations in patient-derived WAS-induced pluripotent stem cell (iPSC) lines, generating isogeneic WAS-iPSC lines. These included lines with direct mutation-specific correction and lines incorporating a WASP transgene cassette regulated by the MND or WAS1.6 kb promoter integrated at the safe harbor AAV1 site. Our results demonstrated that direct mutation correction successfully restored WASP levels to the equivalent of the wild-type in iPSC-derived megakaryocytes (MKs). In contrast, the AAV1-targeted strategy using the MND and WAS1.6 promoters yielded a lower level of WASP. Notably, only the mutation-specific correction lines exhibited improvements in proplatelet structures and generated larger-sized platelets. Our findings underscore the crucial roles of WASP during human thrombopoiesis and suggest that therapeutic approaches, such as direct gene correction, which can achieve physiologic levels of WASP in MKs, hold promise for ameliorating platelet defects in individuals with WAS.

wiskott - aldrich综合征诱导多能干细胞的基因校正修复前血小板缺陷并改善血小板大小。
Wiskott-Aldrich综合征(WAS)是一种严重的x连锁疾病,由WAS基因的功能缺失突变引起,WAS基因负责编码WASP, WASP是除红细胞外所有造血细胞中肌动蛋白细胞骨架的关键调节因子。微血小板减少症是WAS的一个显著特征,也是导致死亡率的一个主要因素,其机制尚未完全阐明。在这项研究中,我们使用不同的基因编辑策略,纠正了患者来源的WAS诱导的多能干细胞系的突变,生成了等基因WAS iPSC系。这些包括具有直接突变特异性纠正的系和包含由MND或在安全港AAV1位点整合的WAS1.6 kb启动子调控的WASP转基因盒的系。我们的研究结果表明,在ipsc衍生的巨核细胞中,直接突变校正成功地将WASP水平恢复到相当于野生型的水平。相比之下,使用MND和WAS1.6启动子的aav1靶向策略产生了较低水平的WASP。值得注意的是,只有突变特异性校正系表现出前血小板结构的改善,并产生更大的血小板。我们的研究结果强调了WASP在人类血小板形成过程中的关键作用,并表明治疗方法,如直接基因校正,可以在巨核细胞中达到生理水平的WASP,有望改善WAS患者的血小板缺陷。
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来源期刊
Thrombosis and haemostasis
Thrombosis and haemostasis 医学-外周血管病
CiteScore
11.90
自引率
9.00%
发文量
140
审稿时长
1 months
期刊介绍: Thrombosis and Haemostasis publishes reports on basic, translational and clinical research dedicated to novel results and highest quality in any area of thrombosis and haemostasis, vascular biology and medicine, inflammation and infection, platelet and leukocyte biology, from genetic, molecular & cellular studies, diagnostic, therapeutic & preventative studies to high-level translational and clinical research. The journal provides position and guideline papers, state-of-the-art papers, expert analysis and commentaries, and dedicated theme issues covering recent developments and key topics in the field.
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