Center for Reproductive Medicine, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, 845 Lingshan Road, Shanghai, 200135, China.
Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Shanghai, China.
J Assist Reprod Genet. 2023 Sep;40(9):2251-2266. doi: 10.1007/s10815-023-02892-y. Epub 2023 Aug 9.
To reveal the underlying roles that pyroptosis-related genes (PRGs) played in human spermatogenic dysfunction.
One discovery set and three validation sets were employed to inspect the previously reported 33 PRGs in the human testis with different status of spermatogenesis. PRGs that differentially expressed in all sets were considered as key differentially expressed pyroptosis-related genes (PR-DEGs). The relationships between key PR-DEGs and samples' clinicopathological, therapeutic, and immune patterns were respectively studied. Single-cell RNA sequencing (scRNS-seq) analyses were conducted to show the expression changes and related mechanisms of key PR-DEGs at a single-cell resolution.
CASP4 and GPX4 were identified as two key PR-DEGs. These two genes were significantly dysregulated in spermatogenic dysfunctional samples, but with opposite tendency. CASP4 was negatively correlated with Johnsen scores but positively correlated with follicle-stimulating hormone (FSH) levels (all p < 0.05), while GPX4 exhibited significant positive correlations with Johnsen scores and negative relevance with FSH. For treatments, both molecules showed a prospective value of being predictors for sperm retrieval surgeries. Moreover, CASP4 and GPX4 were potential immunoregulators in the testicular immune microenvironment and showed significant correlations to testicular macrophages and mast cell infiltration. In scRNA-seq analyses, GPX4 was highly expressed in germ cells, which therefore suffered a sharp reduction with the loss of germ cells in spermatogenic dysfunction. On the other hand, CASP4 were basically somatic cell-derived, and the proportion of CASP4-positive Leydig cells significantly increased in disease testes (p = 0.0001).
In all, we revealed two key PRGs of human testes that might be functional in spermatogenic dysfunction.
揭示与细胞焦亡相关的基因（PRGs）在人类睾丸生殖功能障碍中的潜在作用。
采用一个发现集和三个验证集，分别对不同生精状态的人类睾丸中之前报道的 33 个 PRGs 进行检测。在所有集合中差异表达的 PRGs 被认为是关键差异表达的与细胞焦亡相关基因（PR-DEGs）。分别研究了关键 PR-DEGs 与样本临床病理、治疗和免疫模式之间的关系。进行单细胞 RNA 测序（scRNS-seq）分析，以显示关键 PR-DEGs 在单细胞分辨率下的表达变化及其相关机制。
鉴定出 CASP4 和 GPX4 为两个关键 PR-DEGs。这两个基因在生殖功能障碍样本中明显失调，但趋势相反。Caspase4 与 Johnsen 评分呈负相关，与卵泡刺激素（FSH）水平呈正相关（均 p<0.05），而 GPX4 与 Johnsen 评分呈显著正相关，与 FSH 呈负相关。对于治疗，这两种分子都显示出作为精子提取手术预测因子的前景价值。此外，Caspase4 和 GPX4 是睾丸免疫微环境中的潜在免疫调节剂，与睾丸巨噬细胞和肥大细胞浸润呈显著相关性。在 scRNA-seq 分析中，GPX4 在生殖细胞中高表达，因此在生殖功能障碍时生殖细胞丢失导致其急剧减少。另一方面，Caspase4 基本上来源于体细胞，疾病睾丸中 Caspase4 阳性的间质细胞（Leydig 细胞）的比例显著增加（p=0.0001）。
总之，我们揭示了两个人类睾丸的关键 PRGs，它们可能在生殖功能障碍中具有功能。
