Alfano Massimo, Tascini Anna Sofia, Pederzoli Filippo, Venegoni Chiara, Locatelli Irene, Lesma Arianna, Fallara Giuseppe, Boeri Luca, Pozzi Edoardo, Negri Fausto, Colecchia Maurizio, Pontillo Marina, Montorsi Francesco, Garcia-Manteiga Jose Manuel, Salonia Andrea
Division of Experimental Oncology/Unit of Urology, URI, IRCCS Ospedale San Raffaele, Milan, Italy.
Center for Omics Sciences, IRCCS Ospedale San Raffaele, Milan, Italy.
J Clin Endocrinol Metab. 2025 Jul 14. doi: 10.1210/clinem/dgaf404.
Non-obstructive azoospermia (NOA) is the most severe form of male infertility affecting 1% of all men, with a clinical picture characterized by no sperm production, hyalinization of the basal membrane of the seminiferous tubules, primary hypogonadism and earlier onset of age-related comorbidities compared with fertile men. NOA is also characterized by etiologic heterogeneity and the non-genetic form has higher incidence of testicular germ cell cancer (TGCC) compared to the forms with genetic abnormalities.
We aimed to establish molecular pathways in the testicular somatic cells that are either shared or specific for non-genetic and genetic forms of NOA, as Complete Androgen Insensitivity Syndrome (CAIS) and Klinefelter Syndrome (KS).
Single cell RNAseq of the testicular somatic cells of an individual with CAIS, and data integration with published scRNA-seq datasets of testis with normal spermatogenesis, NOA, KS and germinal testicular cancer. Detailed clinical data of the CAIS patient, Testosterone and Estradiol levels in age-matched men (120 fertile, 155 infertile, 116 NOA, 18 KS, and 343 with TGCC).
In all conditions, Leydig cells are immature and senescent, but those of NOA associated with primary hypogonadism depict the highest expression of transcripts associated with the seminoma microenvironment, including estrogen-responsive genes. An oncological transcriptional signature in the Leydig cells has been confirmed at the systemic levels by showing a prognostic role of the decreasing Testosterone/Estradiol ratio for TGCC in men with non-genetic NOA.
This study offers molecular insights into the prediction of TGCC in persons with NOA and eligibility for the use of aromatase inhibitors.
非梗阻性无精子症（NOA）是男性不育最严重的形式，影响所有男性的1%，临床表现为无精子生成、生精小管基底膜玻璃样变、原发性性腺功能减退，且与有生育能力的男性相比，与年龄相关的合并症发病更早。NOA还具有病因异质性，与有基因异常的形式相比，非基因形式的睾丸生殖细胞癌（TGCC）发病率更高。
我们旨在建立睾丸体细胞中的分子途径，这些途径对于非基因和基因形式的NOA（如完全雄激素不敏感综合征（CAIS）和克兰费尔特综合征（KS））是共享的或特定的。
对一名CAIS个体的睾丸体细胞进行单细胞RNA测序，并将数据与已发表的正常精子发生、NOA、KS和睾丸生殖细胞癌的睾丸单细胞RNA测序数据集进行整合。CAIS患者的详细临床数据、年龄匹配男性（120名有生育能力者、155名不育者、116名NOA患者、18名KS患者和343名TGCC患者）的睾酮和雌二醇水平。
在所有情况下，睾丸间质细胞均不成熟且衰老，但与原发性性腺功能减退相关的NOA患者的睾丸间质细胞中，与精原细胞瘤微环境相关的转录本表达最高，包括雌激素反应基因。通过显示非基因NOA男性中睾酮/雌二醇比值降低对TGCC的预后作用，在系统水平上证实了睾丸间质细胞中的肿瘤转录特征。
本研究为预测NOA患者的TGCC以及使用芳香化酶抑制剂的适用性提供了分子见解。
