In vitro cellular reprogramming to model gonad development and its disorders.
In vitro cellular reprogramming to model gonad development and its disorders.
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DOI：
10.1126/sciadv.abn9793
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发表时间：
2023-01-04
期刊：
影响因子：
13.6
通讯作者：
中科院分区：
文献类型：
--
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在胚胎发育过程中，相互拮抗的信号级联决定了性腺向睾丸或卵巢的命运。这一过程中的错误会导致性发育障碍（DSD），其特征是染色体、性腺和解剖性别之间的不一致。缺乏适当的、可访问的体外系统是理解性别决定/DSD 机制的主要障碍。在这里，我们描述了小鼠和人类多能细胞向性腺祖细胞分化的方案。转录组分析表明，体外衍生的小鼠性腺细胞相当于胚胎第 11.5 天的体内祖细胞。使用类似的条件，源自 46,XY 人类诱导多能干细胞 (hiPSC) 的支持样细胞表现出睾丸特异性基因的持续表达、分泌抗苗勒氏管激素、迁移并形成管状结构。源自 46,XY DSD 雌性 hiPSC 的细胞携带 NR5A1 变体，显示出异常的基因表达和缺乏小管形成。 CRISPR-Cas9 介导的变异校正挽救了表型。这是了解性别决定机制和模型 DSD 的强大工具。建立多能细胞衍生的体外系统来模拟哺乳动物性腺发育和性发育障碍。
During embryonic development, mutually antagonistic signaling cascades determine gonadal fate toward a testicular or ovarian identity. Errors in this process result in disorders of sex development (DSDs), characterized by discordance between chromosomal, gonadal, and anatomical sex. The absence of an appropriate, accessible in vitro system is a major obstacle in understanding mechanisms of sex-determination/DSDs. Here, we describe protocols for differentiation of mouse and human pluripotent cells toward gonadal progenitors. Transcriptomic analysis reveals that the in vitro–derived murine gonadal cells are equivalent to embryonic day 11.5 in vivo progenitors. Using similar conditions, Sertoli-like cells derived from 46,XY human induced pluripotent stem cells (hiPSCs) exhibit sustained expression of testis-specific genes, secrete anti-Müllerian hormone, migrate, and form tubular structures. Cells derived from 46,XY DSD female hiPSCs, carrying an NR5A1 variant, show aberrant gene expression and absence of tubule formation. CRISPR-Cas9–mediated variant correction rescued the phenotype. This is a robust tool to understand mechanisms of sex determination and model DSDs. Pluripotent cell-derived in vitro system is established to model mammalian gonad development and disorders of sex development.
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