Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice.
Precise Correction of Lhcgr Mutation in Stem Leydig Cells by Prime Editing Rescues Hereditary Primary Hypogonadism in Mice.
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DOI：
10.1002/advs.202300993
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发表时间：
2023-10
期刊：
影响因子：
15.1
通讯作者：
Xiang, Andy Peng
中科院分区：
文献类型：
--
作者：
Xia, Kai;Wang, Fulin;Tan, Zhipeng;Zhang, Suyuan;Lai, Xingqiang;Ou, Wangsheng;Yang, Cuifeng;Chen, Hong;Peng, Hao;Luo, Peng;Hu, Anqi;Tu, Xiang'an;Wang, Tao;Ke, Qiong;Deng, Chunhua;Xiang, Andy Peng
遗传性原发性性腺功能减退症（HPH）是由睾丸间质细胞睾酮合成相关基因突变引起的，通常损害男性性发育和精子发生。遗传校正的间质干细胞移植可能为HPH的治疗提供新的途径。在这里，首先基于相对于HPH患者的基因突变产生了一种新的无义点突变小鼠模型（Lhcgr W495X）。为了验证SLC移植治疗HPH的有效性和可行性，将野生型SLC移植到Lhcgr W495 X小鼠体内，其中SLC明显挽救HPH表型。通过比较几种编辑策略，优化的PE2蛋白（PEmax）系统被确定为一个有效的和精确的方法来纠正致病性点突变Lhcgr。此外，通过慢病毒递送内含肽裂解PEmax系统成功地离体校正了来自Lhcgr W495X小鼠的SLC中的突变。来自Lhcgr W495X小鼠的基因校正的SLC在体外发挥分化为功能性Leydig细胞的能力。值得注意的是，基因校正的SLC的移植有效地再生了Leydig细胞，恢复了睾酮的产生，重新启动了性发育，挽救了精子发生，并在Lhcgr W495X小鼠中产生了可生育的后代。总之，这些结果表明，在离体SLC中基于PE的基因编辑是HPH治疗的一种有前途的策略，并可能用于解决生殖系统中更多的遗传性疾病。本研究建立了一种新型遗传性原发性性腺功能减退症小鼠模型，该模型在Lhcgr基因（Lhcgr W495X）中存在无义点突变。优化的PE2蛋白（PEmax）系统是一种有效和精确的方法来纠正Lhcgr W495X小鼠的干Leydig细胞（SLC）中的致病性突变。基因校正的SLC移植恢复了Lhcgr W495X小鼠的睾酮产生，重新启动性发育，挽救精子发生，并产生可生育的后代。
Hereditary primary hypogonadism (HPH), caused by gene mutation related to testosterone synthesis in Leydig cells, usually impairs male sexual development and spermatogenesis. Genetically corrected stem Leydig cells (SLCs) transplantation may provide a new approach for treating HPH. Here, a novel nonsense‐point‐mutation mouse model (Lhcgr W495X) is first generated based on a gene mutation relative to HPH patients. To verify the efficacy and feasibility of SLCs transplantation in treating HPH, wild‐type SLCs are transplanted into Lhcgr W495X mice, in which SLCs obviously rescue HPH phenotypes. Through comparing several editing strategies, optimized PE2 protein (PEmax) system is identified as an efficient and precise approach to correct the pathogenic point mutation in Lhcgr. Furthermore, delivering intein‐split PEmax system via lentivirus successfully corrects the mutation in SLCs from Lhcgr W495X mice ex vivo. Gene‐corrected SLCs from Lhcgr W495X mice exert ability to differentiate into functional Leydig cells in vitro. Notably, the transplantation of gene‐corrected SLCs effectively regenerates Leydig cells, recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in Lhcgr W495X mice. Altogether, these results suggest that PE‐based gene editing in SLCs ex vivo is a promising strategy for HPH therapy and is potentially leveraged to address more hereditary diseases in reproductive system. This study generates a novel hereditary primary hypogonadism mouse model with a nonsense‐point‐mutation in Lhcgr gene (Lhcgr W495X). The optimized PE2 protein (PEmax) system is an efficient and precise approach to correct the pathogenic mutation in stem Leydig cells (SLCs) from Lhcgr W495X mice ex vivo. Transplantation of gene‐corrected SLCs recovers testosterone production, restarts sexual development, rescues spermatogenesis, and produces fertile offspring in Lhcgr W495X mice.
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影响因子：
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作者：
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通讯作者：
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通过表达优化和祖先重建改进胞苷和腺嘌呤碱基编辑器。
影响因子：
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