多组学研究表明，PICK1 缺失通过抑制支持细胞中的囊泡运输导致男性不育。

Multi-omics study identifies that PICK1 deficiency causes male infertility by inhibiting vesicle trafficking in Sertoli cells.

机构信息

Laboratory Medicine Center, Zhejiang Center for Clinical Laboratories, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, 310000, Zhejiang, China.

Laboratory Medicine Center, Allergy Center, Department of Transfusion Medicine, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, 310000, Zhejiang, China.

出版信息

Reprod Biol Endocrinol. 2023 Nov 25;21(1):114. doi: 10.1186/s12958-023-01163-w.

DOI:10.1186/s12958-023-01163-w

PMID:38001535

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10675906/

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.

PURPOSE

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

全球约有 10-15%的育龄男性受到不育症的影响，其中三分之一的病因与遗传因素有关。作为一个 Bin-Amphiphysin-Rvs（BAR）结构域蛋白，蛋白相互作用激酶 1（PICK1）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）的影响尚不清楚。

目的

本研究旨在通过多组学分析研究 PICK1 缺乏对支持细胞的影响，并确定有效的生物标志物，以区分由 PICK1 缺乏引起的不育症男性和生育能力正常的男性。

方法

对 20 名少精症不育症患者进行全外显子测序（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确定致病机制。对支持细胞特异性 PICK1 KO 小鼠进行动物和细胞功能实验，以验证支持细胞的功能障碍。

结果

在患有特定血清抑制素 B 水平降低的不育症患者中发现了 PICK1 中两个导致 BAR 功能结构域转录缺失的无功能缺失突变 c.358delA 和 c.364delA，表明支持细胞功能受损。PICK1 KO 小鼠的多组学分析表明，差异表达 microRNA 和 mRNA 的靶向基因显著富集在囊泡转运途径的负调控作用中，而代谢组学分析显示氨基酸、脂质、辅助因子、维生素和内分泌因子的代谢发生改变。PICK1 KO 小鼠的表型显示睾丸体积减小、成熟精子数量减少以及支持细胞的分泌功能受损。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）的表达减少。