基于微阵列和生物信息学分析鉴定参与非梗阻性无精子症生精细胞和支持细胞的新型细胞骨架蛋白

Identification of novel cytoskeleton protein involved in spermatogenic cells and sertoli cells of non-obstructive azoospermia based on microarray and bioinformatics analysis.

作者信息

Hashemi Karoii Danial, Azizi Hossein, Darvari Maryam, Qorbanee Ali, Hawezy Dawan Jamal

机构信息

Department of Cell and Molecular Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran.

Faculty of Biotechnology, Amol University of Special Modern Technologies, Amol, Iran.

出版信息

BMC Med Genomics. 2025 Jan 25;18(1):19. doi: 10.1186/s12920-025-02087-7.

DOI:10.1186/s12920-025-02087-7

PMID:39863862

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11762539/

Abstract

BACKGROUND

During mammalian spermatogenesis, the cytoskeleton system plays a significant role in morphological changes. Male infertility such as non-obstructive azoospermia (NOA) might be explained by studies of the cytoskeletal system during spermatogenesis.

METHODS

The cytoskeleton, scaffold, and actin-binding genes were analyzed by microarray and bioinformatics (771 spermatogenic cellsgenes and 774 Sertoli cell genes). To validate these findings, we cross-referenced our results with data from a single-cell genomics database.

RESULTS

In the microarray analyses of three human cases with different NOA spermatogenic cells, the expression of TBL3, MAGEA8, KRTAP3-2, KRT35, VCAN, MYO19, FBLN2, SH3RF1, ACTR3B, STRC, THBS4, and CTNND2 were upregulated, while expression of NTN1, ITGA1, GJB1, CAPZA1, SEPTIN8, and GOLGA6L6 were downregulated. There was an increase in KIRREL3, TTLL9, GJA1, ASB1, and RGPD5 expression in the Sertoli cells of three human cases with NOA, whereas expression of DES, EPB41L2, KCTD13, KLHL8, TRIOBP, ECM2, DVL3, ARMC10, KIF23, SNX4, KLHL12, PACSIN2, ANLN, WDR90, STMN1, CYTSA, and LTBP3 were downregulated. A combined analysis of Gene Ontology (GO) and STRING, were used to predict proteins' molecular interactions and then to recognize master pathways. Functional enrichment analysis showed that the biological process (BP) mitotic cytokinesis, cytoskeleton-dependent cytokinesis, and positive regulation of cell-substrate adhesion were significantly associated with differentially expressed genes (DEGs) in spermatogenic cells. Moleculare function (MF) of DEGs that were up/down regulated, it was found that tubulin bindings, gap junction channels, and tripeptide transmembrane transport were more significant in our analysis. An analysis of GO enrichment findings of Sertoli cells showed BP and MF to be common DEGs. Cell-cell junction assembly, cell-matrix adhesion, and regulation of SNARE complex assembly were significantly correlated with common DEGs for BP. In the study of MF, U3 snoRNA binding, and cadherin binding were significantly associated with common DEGs.

CONCLUSION

Our analysis, leveraging single-cell data, substantiated our findings, demonstrating significant alterations in gene expression patterns.

摘要

背景

在哺乳动物精子发生过程中，细胞骨架系统在形态变化中起重要作用。诸如非梗阻性无精子症（NOA）等男性不育症可能通过精子发生过程中细胞骨架系统的研究来解释。

方法

通过微阵列和生物信息学分析细胞骨架、支架和肌动蛋白结合基因（771个生精细胞基因和774个支持细胞基因）。为了验证这些发现，我们将结果与单细胞基因组数据库的数据进行交叉参考。

结果

在对三例不同NOA生精细胞的人类病例进行的微阵列分析中，TBL3、MAGEA8、KRTAP3-2、KRT35、VCAN、MYO19、FBLN2、SH3RF1、ACTR3B、STR C、THBS4和CTNND2的表达上调，而NTN1、ITGA1、GJB1、CAPZA1、SEPTIN8和GOLGA6L6的表达下调。在三例NOA人类病例的支持细胞中，KIRREL3、TTLL9、GJA1、ASB1和RGPD5的表达增加，而DES、EPB41L2、KCTD13、KLHL8、TRIOBP、ECM2、DVL3、ARMC10、KIF23、SNX4、KLHL12、PACSIN2、ANLN、WDR90、STMN1、CYTSA和LTBP3的表达下调。基因本体论（GO）和STRING的联合分析用于预测蛋白质的分子相互作用，进而识别主要途径。功能富集分析表明，生物学过程（BP）有丝分裂胞质分裂、细胞骨架依赖性胞质分裂和细胞-底物粘附的正调控与生精细胞中差异表达基因（DEG）显著相关。对上调/下调的DEG的分子功能（MF）分析发现，微管蛋白结合、间隙连接通道和三肽跨膜转运在我们的分析中更显著。对支持细胞的GO富集结果分析表明，BP和MF是常见的DEG。细胞-细胞连接组装、细胞-基质粘附和SNARE复合体组装的调控与BP的常见DEG显著相关。在MF研究中，U3小核仁RNA结合和钙粘蛋白结合与常见DEG显著相关。