High-fidelity reprogramming into Leydig-like cells by CRISPR activation and paracrine factors.
High-fidelity reprogramming into Leydig-like cells by CRISPR activation and paracrine factors.
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通过 CRISPR 激活和旁分泌因子高保真重编程为 Leydig 样细胞。
DOI：
10.1093/pnasnexus/pgac179
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发表时间：
2022-09
期刊：
影响因子：
--
通讯作者：
Su, Zhijian
中科院分区：
文献类型：
--
作者：
Li, Zhaohui;Fan, Yuxiao;Xie, Cankun;Liu, Jierong;Guan, Xiaoju;Li, Shijun;Huang, Yadong;Zeng, Rong;Chen, Haolin;Su, Zhijian
摘要雄激素缺乏是一种影响所有年龄段男性的常见疾病。睾酮产生细胞移植是治疗男性性腺功能减退的一种很有前途的方法。然而，获得具有间质细胞特性的细胞来源仍然是一个挑战。本文描述了基于表观遗传机制的皮肤源性成纤维细胞高效重编程为功能性leydigi样细胞（LLCs）。通过对LCs和成纤维细胞的全基因组DNA甲基化和转录组谱进行综合分析，确定了潜在的表观遗传调节类固醇基因和信号通路。然后利用CRISPR/dCas9激活系统和信号通路调节剂，将雄性或雌性来源的成纤维细胞重编程为具有主要lc特异性特征的LLCs。转录组学分析进一步表明，LLCs和LCs之间的全局基因和转录因子的相关系数分别大于0.81和0.96。移植到性腺功能低下的啮齿动物模型睾丸后，LLCs显著提高了血清睾酮浓度。在2型糖尿病大鼠模型中，腋窝移植LLCs能够恢复血清睾酮水平，改善高血糖状态。总之，我们的方法使皮肤来源的成纤维细胞能够高保真地重编程为llc，为男性性腺功能减退和代谢相关合并症的治疗提供了潜在的细胞来源。
Abstract Androgen deficiency is a common medical conditions that affects males of all ages. Transplantation of testosterone-producing cells is a promising treatment for male hypogonadism. However, getting a cell source with the characteristics of Leydig cells (LCs) is still a challenge. Here, a high-efficiency reprogramming of skin-derived fibroblasts into functional Leydig-like cells (LLCs) based on epigenetic mechanism was described. By performing an integrated analysis of genome-wide DNA methylation and transcriptome profiling in LCs and fibroblasts, the potentially epigenetic-regulating steroidogenic genes and signaling pathways were identified. Then by using CRISPR/dCas9 activation system and signaling pathway regulators, the male- or female-derived fibroblasts were reprogrammed into LLCs with main LC-specific traits. Transcriptomic analysis further indicated that the correlation coefficients of global genes and transcription factors between LLCs and LCs were higher than 0.81 and 0.96, respectively. After transplantation in the testes of hypogonadal rodent models, LLCs increased serum testosterone concentration significantly. In type 2 diabetic rats model, LLCs which were transplanted in armpit, have the capability to restore the serum testosterone level and improve the hyperglycemia status. In conclusion, our approach enables skin-derived fibroblasts reprogramming into LLCs with high fidelity, providing a potential cell source for the therapeutics of male hypogonadism and metabolic-related comorbidities.
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影响因子：
4.1
作者：
Chen H;Ge RS;Zirkin BR
通讯作者：
Zirkin BR
DOI：
10.1621/nrs.11001
发表时间：
2013
期刊：
Nuclear receptor signaling
影响因子：
--
作者：
Chang C;Yeh S;Lee SO;Chang TM
通讯作者：
Chang TM
影响因子：
2.9
作者：
通讯作者：
--
DOI：
10.1073/pnas.1908207116
发表时间：
2019-11-12
影响因子：
11.1
作者：
Li, Lu;Li, Yuchang;Papadopoulos, Vassilios
通讯作者：
Papadopoulos, Vassilios
影响因子：
5.8
作者：
Bhasin, Shalender;Brito, Juan P.;Yialamas, Maria A.
通讯作者：
Yialamas, Maria A.