In some women, sexually transmittedChlamydia trachomatismay ascend to infect the endometrium, leading to pelvic inflammatory disease. To identify endometrial innate immune components that interact withChlamydia, we introducedC. trachomatisinto mouse endometrium via transcervical inoculation and compared the infectious yields in mice with and without immunodeficiency. LiveC. trachomatisrecovered from vaginal swabs or endometrial tissues peaked on day 3 and then declined in all mice with or without deficiency in adaptive immunity, indicating a critical role for innate immunity in endometrial control ofC. trachomatisinfection.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder affecting women of reproductive age. It is characterized by symptoms such as hyperandrogenemia, oligo or anovulation and polycystic ovarian, significantly impacting quality of life. However, the practical implementation of machine learning (ML) in PCOS diagnosis is hindered by the limitations related to data size and algorithmic models. To address this research gap, we have increased the sample size in our study and aim to utilize two ML algorithms to analyze and validate diagnostic biomarkers, as well as explore immune cell infiltration patterns in PCOS. We performed RNA-seq analysis on granulosa cell, including 13 samples from normal controls and 25 samples from women with PCOS. The data from our study were combined with publicly available databases. Batch effects were corrected using the ‘sva’ package in R software. Differential expression analysis was performed to identify genes that exhibited significant differences between the two groups. These differentially expressed genes (DEGs) were further analyzed for Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Hub genes were selected by intersecting the results of both methods after using LASSO and SVM-RFE for central gene selection for DEGs. Receiver Operating Characteristic (ROC) curves were employed to verify the accuracy of models by SVM and XGBoost. CIBERSORT analysis was performed to determine the relative abundances of immune cell populations. GSEA was analyzed to illustrate the expression patterns of genes within highly enriched functional pathways. RT-qPCR was used to validate the reliability of hub genes. 824 DEGs were found between the normal control and PCOS groups, including 376 upregulated and 448 downregulated genes. These DEGs were associated with endocytosis, salmonella infection and focal adhesion based on the KEGG enrichment analysis. Through overlapping LASSO and SVM-RFE algorithms, we identified four hub genes (CNTN2, CASR, CACNB3, MFAP2) that are significantly associated with the PCOS group. The diagnostic efficacy validation set using SVM and XGBoost yielded AUC values of 0.795 and 0.875, respectively, indicating their potential as diagnostic biomarkers. Consistent with the data analysis, the upregulation of CNTN2, CASR, CACNB3, and MFAP2 in PCOS was confirmed by RT-qPCR analysis on human granulosa cells. Furthermore, according to CIBERSORT analysis, a significant reduction in CD4 memory resting T cells was revealed in the PCOS group compared to the normal control group (P < 0.05). This study identified CNTN2, CASR, CACNB3, and MFAP2 as potential diagnostic biomarkers for PCOS, which provides strong evidence for existing research on hub genes. Furthermore, the analysis of immune cell infiltration revealed the significant involvement of CD4 memory resting T cells in the onset and progression of PCOS. These findings shed light on potential mechanisms underlying PCOS pathogenesis and provide valuable insights for future research and therapeutic interventions.

Premature ovarian insufficiency (POI) is characterized by ovarian dysfunction that develops from diminished ovarian reserve (DOR). The exact aetiology of POI remains poorly understood. This study aims to elucidate the role of CKAP5 in the regulation of ovarian function and fertility. Bulk RNA sequencing of granulosa cells was conducted in the control group and in the patients with DOR to screen for candidate genes, which were further validated by gene burden analysis in a next-generation sequencing cohort of POI and control individuals. Additionally, ovarian reserve was evaluated in heterozygous Ckap5 knockout mice, alongside the ovarian and oocyte single-cell transcriptome analysis. The regulatory mechanism of CKAP5 was studied through in vivo and in vitro experiments. CKAP5 was identified as a key hub gene associated with ovarian ageing. Heterozygous Ckap5 knockout mice exhibited a POI-like phenotype, characterized by a reduced primordial follicle pool and accelerated follicular atresia. CKAP5 promotes autophagy via ATG7 and simultaneously supports DNA damage repair through the ATM. Finally, a variant in CKAP5 (NM_0001008938.4, c.630 + 7_630 + 11delCAAAA) was identified in patients with POI, resulting in protein truncation and loss of function. CKAP5 deficiency induces premature ovarian insufficiency in both humans and mice. The National Key R&D Program of China (2017YFC1001100), the National Natural Science Foundation of China (81501248, 81471453 and 81801295), the Health Research Project of Hunan Provincial Health Commission (W20243018), the Science and Technology Innovation Program of Hunan Province (2021RC3031), the National Natural Science Foundation of Hunan Province (2022JJ30066), the Scientific Research Program of Hunan Provincial Health Commission (202205033471 and 21B0058), the Open Research Fund of Hunan Provincial Key Laboratory of Regional Hereditary Birth Defects Prevention and Control (HPKL2023013).

Sexually transmitted Chlamydia trachomatis can ascend to the upper genital tract due to its resistance to innate immunity in the lower genital tract. C. trachomatis can activate the cGAS-STING signaling pathway in cultured cells via either cGAS or STING. This study was designed to evaluate the role of the cGAS-STING pathway in innate immunity against C. trachomatis in the mouse genital tract.