DDX3Y gene rescue of a Y chromosome AZFa deletion restores germ cell formation and transcriptional programs.
DDX3Y gene rescue of a Y chromosome AZFa deletion restores germ cell formation and transcriptional programs.
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DOI：
10.1038/srep15041
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发表时间：
2015-10-12
影响因子：
4.6
通讯作者：
Reijo Pera R
中科院分区：
文献类型：
--
作者：
Ramathal C;Angulo B;Sukhwani M;Cui J;Durruthy-Durruthy J;Fang F;Schanes P;Turek PJ;Orwig KE;Reijo Pera R
人类Y染色体的AZFa区域（A精子因子-a）区域的缺失导致不可逆的生精失败，其在临床上在男性中表现为睾丸的仅支持细胞（SCO）病理学。AZFa区域的缺失通常包括两个基因：DDX 3 Y和USP 9 Y。然而，人类遗传学证据表明，SCO与DDX 3 Y的缺失联系最紧密，并且USP 9 Y的缺失/突变可以从一代传递到下一代。在这里，我们产生了具有AZFa缺失的稳定iPSC系，通过引入DDX 3 Y测试了互补，并评估了在异种移植模型中体内形成生殖细胞的能力。我们观察到从补充的供体iPSC形成生殖细胞样细胞（GCLC）的可量化的改善。此外，通过在AZFa背景上引入DDX 3 Y补充，UTF 1（一种促性腺激素蛋白）的表达在细胞中得以恢复。纯化的GCLC的全基因组RNA测序揭示了DDX 3 Y拯救的GCLC中参与翻译抑制和转录控制的基因相对于突变GCLC的富集，突变GCLC保持了与未分化的iPSC更相似的分子表型。这项研究证明了通过互补来探索人类生殖细胞形成的基础遗传学的能力，并表明DDX 3 Y在人类生殖细胞发育的最早阶段发挥作用。
Deletions of the AZFa region (AZoospermia Factor-a) region of the human Y chromosome cause irreversible spermatogenic failure that presents clinically in men as Sertoli-cell only (SCO) pathology of the testis. Deletions of the AZFa region typically encompass two genes: DDX3Y and USP9Y. However, human genetic evidence indicates that SCO is most tightly linked to deletion of DDX3Y and that deletions/mutations of USP9Y can be transmitted from one generation to the next. Here, we generated stable iPSC lines with AZFa deletions, tested complementation via introduction of DDX3Y, and assessed ability to form germ cells in vivo in a xenotransplantation model. We observed a quantifiable improvement in formation of germ cell like cells (GCLCs) from complemented donor iPSCs. Moreover, expression of UTF1, a prospermatogonial protein, was restored in cells complemented by introduction of DDX3Y on the AZFa background. Whole-genome RNA sequencing of purified GCLCs revealed an enrichment of genes involved in translational suppression and transcriptional control in DDX3Y-rescued GCLCs over mutant GCLCs, which maintained a molecular phenotype more similar to undifferentiated iPSCs. This study demonstrates the ability to probe fundamental genetics of human germ cell formation by complementation and indicates that DDX3Y functions in the earliest stages of human germ cell development.