Apart from the androgen receptor, transcription factors (TFs) that are required for the development and formation of the different segments of the epididymis have remained unknown. We identified TF families expressed in the developing epididymides, of which many showed segment specificity. From these TFs, down-regulation of runt related transcription factors (RUNXs) 1 and 2 expression coincides with epithelial regression in Dicer1 cKO mice. Concomitant deletion of both Runx1 and Runx2 in a mouse epididymal epithelial cell line affected cell morphology, adhesion and mobility in vitro. Furthermore, lack of functional RUNXs severely disturbed the formation of 3D epididymal organoid-like structures. Transcriptomic analysis of the epididymal cell organoid-like structures indicated that RUNX1 and RUNX2 are involved in the regulation of MAPK signaling, NOTCH pathway activity, and EMT-related gene expression. This suggests that RUNXs are master regulators of several essential signaling pathways, and necessary for the maintenance of proper differentiation of the epididymal epithelium.

除了雄激素受体外，附睾不同节段的发育和形成所需的转录因子（TF）仍然未知。我们鉴定了在发育中的附睾中表达的 TF 家族，其中许多家族表现出片段特异性。在这些 TF 中，矮小相关转录因子 (RUNX) 1 和 2 表达的下调与 Dicer1 cKO 小鼠的上皮退化一致。在小鼠附睾上皮细胞系中同时删除Runx1和Runx2会影响体外细胞的形态、粘附和迁移率。此外，功能性 RUNX 的缺乏严重干扰了 3D 附睾类器官结构的形成。附睾细胞类器官结构的转录组分析表明，RUNX1和RUNX2参与MAPK信号传导、NOTCH通路活性和EMT相关基因表达的调节。这表明 RUNX 是几个重要信号通路的主要调节因子，对于维持附睾上皮的正确分化是必需的。