Deciphering the autophagy regulatory network via single-cell transcriptome analysis reveals a requirement for autophagy homeostasis in spermatogenesis.
Deciphering the autophagy regulatory network via single-cell transcriptome analysis reveals a requirement for autophagy homeostasis in spermatogenesis.
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通过单细胞转录组分析破译自噬调节网络揭示了精子发生中自噬稳态的要求
DOI：
10.7150/thno.55645
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发表时间：
2021
期刊：
影响因子：
12.4
通讯作者：
Zhao XY
中科院分区：
文献类型：
--
作者：
Wang M;Xu Y;Zhang Y;Chen Y;Chang G;An G;Yang X;Zheng C;Zhao J;Liu Z;Wang D;Miao K;Rao S;Dai M;Wang D;Zhao XY
背景资料：自噬是精子发生的重要组成部分，在动物模型中，自噬功能障碍可导致生殖障碍，包括酵母、C。线虫和老鼠然而，复杂的转录网络的自噬基因在整个人类精子发生和它们的生物学意义仍然在很大程度上是未知的。研究方法：我们分析了人类精子发生过程中自噬相关基因的转录特征，通过评估来自9名生育对照（包括2名正常人和7名阻塞性无精子症（OA）患者）和1名非阻塞性无精子症（NOA）患者的标本，使用单细胞RNA测序（scRNA-seq）分析。通过免疫荧光染色证实了另外两名NOA患者的自噬失调。基因敲除用于鉴定Cst 3在精子发生期间自噬中的作用。结果：我们的数据揭示了自噬相关基因的独特的、全球性的阶段特异性富集。人-鼠比较分析表明，自噬相关基因的阶段特异性表达模式在哺乳动物中高度保守。更重要的是，在NOA患者中观察到一些自噬相关基因簇的失调，表明自噬与男性不育有关。Cst 3是一个在精原细胞和早期精母细胞中活跃表达的人-小鼠保守的自噬相关基因，被发现调节精原干细胞（SSC）的维持和随后的雄性生殖细胞发育。敲低Cst 3增加了小鼠SSC中的自噬活性，随后抑制了SSC核心因子如Oct 4、Id 1和Nanos 3的转录，这些因子可以通过操纵自噬活性来有效地拯救。结论：我们的研究为自噬相关基因的全球转录特征提供了全面的见解，并证实了自噬稳态在SSC维持和正常精子发生中的重要性，为进一步剖析自噬调控网络在精子发生和男性不育中的意义开辟了新的途径。
Background: Autophagy has been implicated as a crucial component in spermatogenesis, and autophagy dysfunction can lead to reproductive disorders in animal models, including yeast, C. elegans and mice. However, the sophisticated transcriptional networks of autophagic genes throughout human spermatogenesis and their biological significance remain largely uncharacterized. Methods: We profiled the transcriptional signatures of autophagy-related genes during human spermatogenesis by assessing specimens from nine fertile controls (including two normal persons and seven obstructive azoospermia (OA) patients) and one nonobstructive azoospermia (NOA) patient using single-cell RNA sequencing (scRNA-seq) analysis. Dysregulation of autophagy was confirmed in two additional NOA patients by immunofluorescence staining. Gene knockdown was used to identify the role of Cst3 in autophagy during spermatogenesis. Results: Our data uncovered a unique, global stage-specific enrichment of autophagy-related genes. Human-mouse comparison analysis revealed that the stage-specific expression pattern of autophagy-related genes was highly conserved in mammals. More importantly, dysregulation of some clusters of autophagy-related genes was observed in NOA patients, suggesting the association of autophagy with male infertility. Cst3, a human-mouse conserved and autophagy-related gene that is actively expressed in spermatogonia and early spermatocytes, was found to regulate spermatogonial stem cell (SSC) maintenance and subsequent male germ cell development. Knockdown of Cst3 increased autophagic activity in mouse SSCs and subsequently suppressed the transcription of SSC core factors such as Oct4, Id1, and Nanos3, which could be efficiently rescued by manipulating autophagic activity. Conclusions: Our study provides comprehensive insights into the global transcriptional signatures of autophagy-related genes and confirms the importance of autophagy homeostasis in SSC maintenance and normal spermatogenesis, opening new avenues for further dissecting the significance of the autophagy regulatory network in spermatogenesis as well as male infertility.
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影响因子：
13.3
作者：
Da Ros M;Lehtiniemi T;Olotu O;Fischer D;Zhang FP;Vihinen H;Jokitalo E;Sironen A;Toppari J;Kotaja N
通讯作者：
Kotaja N
影响因子：
12.3
作者：
Dong J;Hu Y;Fan X;Wu X;Mao Y;Hu B;Guo H;Wen L;Tang F
通讯作者：
Tang F
影响因子：
11.8
作者：
Green CD;Ma Q;Manske GL;Shami AN;Zheng X;Marini S;Moritz L;Sultan C;Gurczynski SJ;Moore BB;Tallquist MD;Li JZ;Hammoud SS
通讯作者：
Hammoud SS
影响因子：
23.9
作者：
Guo J;Grow EJ;Yi C;Mlcochova H;Maher GJ;Lindskog C;Murphy PJ;Wike CL;Carrell DT;Goriely A;Hotaling JM;Cairns BR
通讯作者：
Cairns BR
自噬核心蛋白 ATG5 是延长雄性小鼠精子细胞发育、精子个体化和正常生育能力所必需的
影响因子：
13.3
作者：
Huang, Qian;Liu, Yunhao;Zhang, Zhibing
通讯作者：
Zhang, Zhibing