PDF(Pubmed)
Abstract：
BACKGROUND: Infertility affects approximately 10-15% of reproductive-age men worldwide, and genetic causes play a role in one-third of cases. As a Bin-Amphiphysin-Rvs (BAR) domain protein, protein interacting with C-kinase 1 (PICK1) deficiency could lead to impairment of acrosome maturation. However, its effects on auxiliary germ cells such as Sertoli cells are unknown.
OBJECTIVE: The present work was aimed to use multi-omics analysis to research the effects of PICK1 deficiency on Sertoli cells and to identify effective biomarkers to distinguish fertile males from infertile males caused by PICK1 deficiency.
METHODS: Whole-exome sequencing (WES) was performed on 20 infertility patients with oligozoospermia to identify pathogenic PICK1 mutations. Multi-omics analysis of a PICK1 knockout (KO) mouse model was utilized to identify pathogenic mechanism. Animal and cell function experiments of Sertoli cell-specific PICK1 KO mouse were performed to verify the functional impairment of Sertoli cells.
RESULTS: Two loss-of-function deletion mutations c.358delA and c.364delA in PICK1 resulting in transcription loss of BAR functional domain were identified in infertility patients with a specific decrease in serum inhibin B, indicating functional impairment of Sertoli cells. Multi-omics analysis of PICK1 KO mouse illustrated that targeted genes of differentially expressed microRNAs and mRNAs are significantly enriched in the negative regulatory role in the vesicle trafficking pathway, while metabolomics analysis showed that the metabolism of amino acids, lipids, cofactors, vitamins, and endocrine factors changed. The phenotype of PICK1 KO mouse showed a reduction in testis volume, a decreased number of mature spermatozoa and impaired secretory function of Sertoli cells. In vitro experiments confirmed that the expression of growth factors secreted by Sertoli cells in PICK1 conditional KO mouse such as Bone morphogenetic protein 4 (BMP4) and Fibroblast growth factor 2 (FGF2) were decreased.
CONCLUSIONS: Our study attributed male infertility caused by PICK1 deficiency to impaired vesicle-related secretory function of Sertoli cells and identified a variety of significant candidate biomarkers for male infertility induced by PICK1 deficiency.
OBJECTIVE: The present work was aimed to use multi-omics analysis to research the effects of PICK1 deficiency on Sertoli cells and to identify effective biomarkers to distinguish fertile males from infertile males caused by PICK1 deficiency.
METHODS: Whole-exome sequencing (WES) was performed on 20 infertility patients with oligozoospermia to identify pathogenic PICK1 mutations. Multi-omics analysis of a PICK1 knockout (KO) mouse model was utilized to identify pathogenic mechanism. Animal and cell function experiments of Sertoli cell-specific PICK1 KO mouse were performed to verify the functional impairment of Sertoli cells.
RESULTS: Two loss-of-function deletion mutations c.358delA and c.364delA in PICK1 resulting in transcription loss of BAR functional domain were identified in infertility patients with a specific decrease in serum inhibin B, indicating functional impairment of Sertoli cells. Multi-omics analysis of PICK1 KO mouse illustrated that targeted genes of differentially expressed microRNAs and mRNAs are significantly enriched in the negative regulatory role in the vesicle trafficking pathway, while metabolomics analysis showed that the metabolism of amino acids, lipids, cofactors, vitamins, and endocrine factors changed. The phenotype of PICK1 KO mouse showed a reduction in testis volume, a decreased number of mature spermatozoa and impaired secretory function of Sertoli cells. In vitro experiments confirmed that the expression of growth factors secreted by Sertoli cells in PICK1 conditional KO mouse such as Bone morphogenetic protein 4 (BMP4) and Fibroblast growth factor 2 (FGF2) were decreased.
CONCLUSIONS: Our study attributed male infertility caused by PICK1 deficiency to impaired vesicle-related secretory function of Sertoli cells and identified a variety of significant candidate biomarkers for male infertility induced by PICK1 deficiency.
摘要：
背景：全世界约有10-15%的育龄男性受不孕症影响，遗传原因在三分之一的病例中起作用。作为Bin-Amphiphysin-Rvs（BAR）结构域蛋白，与C激酶1（PICK1）相互作用的蛋白质缺乏可能导致顶体成熟受损。然而,其对辅助生殖细胞如支持细胞的影响是未知的。
目的：本研究的目的是利用多组学分析来研究PICK1缺乏对支持细胞的影响，并确定有效的生物标志物来区分PICK1缺乏引起的育性男性和不育男性。
方法：对20例少精子症不育患者进行全外显子组测序（WES），以鉴定致病性PICK1突变。利用PICK1敲除(KO)小鼠模型的多组学分析来鉴定致病机制。对Sertoli细胞特异性PICK1KO小鼠进行动物和细胞功能实验以验证Sertoli细胞的功能损害。
结果：在不孕症患者中发现了导致BAR功能域转录丢失的PICK1中的两个功能缺失缺失突变c.358delA和c.364delA，提示支持细胞功能受损。对PICK1KO小鼠的多组学分析表明，差异表达的microRNAs和mRNAs的靶向基因在囊泡运输途径中具有明显的负调节作用，代谢组学分析表明,氨基酸的代谢,脂质，辅因子，维生素,内分泌因素发生了变化。PICK1KO小鼠的表型显示睾丸体积减少，成熟精子数量减少和支持细胞分泌功能受损。体外实验证实，PICK1条件性KO小鼠支持细胞分泌的生长因子如骨形态发生蛋白4（BMP4）和成纤维细胞生长因子2（FGF2）的表达降低。
结论：我们的研究将PICK1缺乏导致的男性不育归因于支持细胞的囊泡相关分泌功能受损，并确定了多种由PICK1缺乏导致的男性不育的重要候选生物标志物。
目的：本研究的目的是利用多组学分析来研究PICK1缺乏对支持细胞的影响，并确定有效的生物标志物来区分PICK1缺乏引起的育性男性和不育男性。
方法：对20例少精子症不育患者进行全外显子组测序（WES），以鉴定致病性PICK1突变。利用PICK1敲除(KO)小鼠模型的多组学分析来鉴定致病机制。对Sertoli细胞特异性PICK1KO小鼠进行动物和细胞功能实验以验证Sertoli细胞的功能损害。
结果：在不孕症患者中发现了导致BAR功能域转录丢失的PICK1中的两个功能缺失缺失突变c.358delA和c.364delA，提示支持细胞功能受损。对PICK1KO小鼠的多组学分析表明，差异表达的microRNAs和mRNAs的靶向基因在囊泡运输途径中具有明显的负调节作用，代谢组学分析表明,氨基酸的代谢,脂质，辅因子，维生素,内分泌因素发生了变化。PICK1KO小鼠的表型显示睾丸体积减少，成熟精子数量减少和支持细胞分泌功能受损。体外实验证实，PICK1条件性KO小鼠支持细胞分泌的生长因子如骨形态发生蛋白4（BMP4）和成纤维细胞生长因子2（FGF2）的表达降低。
结论：我们的研究将PICK1缺乏导致的男性不育归因于支持细胞的囊泡相关分泌功能受损，并确定了多种由PICK1缺乏导致的男性不育的重要候选生物标志物。
