A novel experimental <i>in vitro</i> human definitive endoderm cell-derived model of Wilson disease

Q4 Medicine

Genes and Cells Pub Date : 2025-09-03 DOI:10.17816/gc677378

M. A. Berestovoy, Varvara D. Starodubova, Elizaveta V. Karpukhina, Arina S. Baranova, Arina G. Shokhina

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引用次数: 0

Abstract

BACKGROUND: Wilson disease is a rare autosomal recessive disorder involving mutations in the ATP7B gene, which encodes the copper-transporting ATPase. The dysfunctional protein disrupts biliary copper excretion, which results in copper accumulation in hepatocytes. The available range of cell models to investigate the molecular mechanisms of this disease and to identify novel therapeutic approaches is currently limited. AIM: To develop an in vitro induced human pluripotent stem cell-derived model for investigating the molecular mechanisms of Wilson disease and evaluating therapeutic strategies. METHODS: A 2D cell model has been developed using validated induced pluripotent definitive endoderm cells obtained from a healthy donor and differentiated by activin A and CHIR99021. The copper overload that is a hallmark of the pathogenesis of Wilson disease was simulated by introducing exogenous copper into the growth medium. Relative cell viability was measured by the Alamar Blue assay. RESULTS: The induced pluripotent stem cells demonstrate a normal karyotype. Their morphology is typical of embryonic stem cells. They express pluripotency markers (SOX2, OCT4, TRA-1-60, and SSEA-4) and form tissues of all three germ layers during spontaneous differentiation in embryoid bodies. The differentiation of these induced pluripotent stem cells using the suggested procedure produces definitive endoderm cells that exhibit the expected morphology and express the markers SOX17, FOXA2, and ATP7B. The obtained model demonstrates sensitivity to exogenous copper overload at IC50 197 μM. CONCLUSION: The developed platform of definitive endoderm cells obtained from healthy donor’s induced pluripotent stem cells can be used to model the copper overload in vitro, simulating a cellular metabolic dysfunction associated with Wilson disease. Therefore, the proposed model can be used for both fundamental research and the development of novel therapeutic approaches.

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一种新的实验性体外肝豆状核变性人最终内胚层细胞衍生模型

背景：威尔逊病是一种罕见的常染色体隐性遗传病，涉及编码铜转运atp酶的ATP7B基因突变。功能失调的蛋白质破坏胆铜排泄，导致铜在肝细胞中积聚。目前，用于研究这种疾病的分子机制和确定新的治疗方法的细胞模型的可用范围有限。目的：建立体外诱导的人多能干细胞衍生模型，用于研究肝豆状核变性的分子机制和评估治疗策略。方法：利用从健康供体获得的经过验证的诱导多能最终内胚层细胞，并通过激活素A和CHIR99021分化，建立了2D细胞模型。铜超载是威尔逊病发病机制的一个标志，通过向生长培养基中引入外源铜来模拟。用Alamar Blue法测定相对细胞活力。结果：诱导多能干细胞核型正常。它们的形态是典型的胚胎干细胞。它们表达多能性标记（SOX2、OCT4、TRA-1-60和SSEA-4），并在胚状体自发分化过程中形成所有三个胚层的组织。这些诱导多能干细胞的分化使用建议的程序产生最终的内胚层细胞，表现出预期的形态和表达标记SOX17， FOXA2和ATP7B。所获得的模型在IC50 197 μM下对外源铜过载具有敏感性。结论：建立的健康供体诱导多能干细胞终代内胚层细胞平台可用于体外铜超载模型，模拟与Wilson病相关的细胞代谢功能障碍。因此，所提出的模型既可用于基础研究，也可用于开发新的治疗方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Genes and Cells Medicine-Transplantation

CiteScore

0.50

自引率

0.00%

发文量

期刊介绍： “Genes and Cells” (the old name is “Cell Transplantology and Tissue Engineering”) is a peer-reviewed scientific and practical journal recommended by the Higher Attestation Commission of the Ministry of Education and Science of the Russian Federation for publishing basic materials of dissertation research. Originally conceived as a highly specialized publication, the Journal has now gained an ever wider target audience. If at the beginning of its journey — from September 2005 — the target audience of the journal were biotechnologists, specialists in the field of molecular and cell biology, by now it has expanded with medical practitioners. Such progressive dynamics are absolutely natural — over the last 7-10 years, biomedical technologies have come out of the exotic category, lack of understanding of the inhabitants and mistrust of clinicians have been replaced by interest and awareness of the need to use biotechnology tools in medicine no longer tomorrow, but today. The sections of the journal are formulated to fully disclose the target topics of the publication, convey to readers the opinions of leading experts in the field of biomedical technologies on topical issues of concern, acquaint them with the most significant recent foreign and domestic research, materials of thematic conferences, present analytical information on fundamental issues of biomedical technologies trends in the biotech business. The journal includes the following headings: “expert opinions”, “cell technology news”, “reviews”, “original research”, “clinical experience”, “discussion and general theoretical work”, “stem cell business”.