Experimental Hepatology Unit, Instituto de Investigación Sanitaria La Fe, CIBERehd, Hospital Universitari i Politècnic La Fe, Avda. Fernando Abril Martorell 106, 46026, Valencia, Spain.
Biochemistry and Molecular Biology Department, Universidad de Valencia, Valencia, Spain.
Sci Rep. 2022 Feb 10;12(1):2308. doi: 10.1038/s41598-022-06184-w.
Autologous cell replacement therapy for inherited metabolic disorders requires the correction of the underlying genetic mutation in patient's cells. An unexplored alternative for females affected from X-linked diseases is the clonal selection of cells randomly silencing the X-chromosome containing the mutant allele, without in vivo or ex vivo genome editing. In this report, we have isolated dermal fibroblasts from a female patient affected of ornithine transcarbamylase deficiency and obtained clones based on inactivation status of either maternally or paternally inherited X chromosome, followed by differentiation to hepatocytes. Hepatocyte-like cells derived from these clones display indistinct features characteristic of hepatocytes, but express either the mutant or wild type OTC allele depending on X-inactivation pattern. When clonally derived hepatocyte-like cells were transplanted into FRG KO mice, they were able to colonize the liver and recapitulate OTC-dependent phenotype conditioned by X-chromosome inactivation pattern. This approach opens new strategies for cell therapy of X-linked metabolic diseases and experimental in vitro models for drug development for such diseases.
自体细胞替代疗法治疗遗传性代谢疾病需要纠正患者细胞中潜在的基因突变。对于 X 连锁疾病的女性患者，一种尚未被探索的替代方法是随机选择沉默携带突变等位基因的 X 染色体的细胞进行克隆选择，而无需体内或体外基因组编辑。在本报告中，我们从一名患有鸟氨酸转氨甲酰酶缺乏症的女性患者中分离出皮肤成纤维细胞，并根据母源或父源 X 染色体失活状态获得克隆，然后分化为肝细胞。这些克隆衍生的类肝细胞表现出明显不同于肝细胞的特征，但根据 X 染色体失活模式表达突变型或野生型 OTC 等位基因。当克隆衍生的类肝细胞被移植到 FRG KO 小鼠中时，它们能够定植于肝脏，并重现由 X 染色体失活模式决定的 OTC 依赖性表型。这种方法为 X 连锁代谢疾病的细胞治疗和此类疾病的药物开发提供了新的实验体外模型。
