Essential Roles of Cyclin Y-Like 1 and Cyclin Y in Dividing Wnt-Responsive Mammary Stem/Progenitor Cells

Cyclin Y family can enhance Wnt/β-catenin signaling in mitosis. Their physiological roles in mammalian development are yet unknown. Here we show that Cyclin Y-like 1 (Ccnyl1) and Cyclin Y (Ccny) have overlapping function and are crucial for mouse embryonic development and mammary stem/progenitor cell functions. Double knockout of Ccnys results in embryonic lethality at E16.5. In pubertal development, mammary terminal end buds robustly express Ccnyl1. Depletion of Ccnys leads to reduction of Lrp6 phosphorylation, hampering β-catenin activities and abolishing mammary stem/progenitor cell expansion in vitro. In lineage tracing experiments, Ccnys-deficient mammary cells lose their competitiveness and cease to contribute to mammary development. In transplantation assays, Ccnys-deficient mammary cells fail to reconstitute, whereas constitutively active β-catenin restores their regeneration abilities. Together, our results demonstrate the physiological significance of Ccnys-mediated mitotic Wnt signaling in embryonic development and mammary stem/progenitor cells, and reveal insights in the molecular mechanisms orchestrating cell cycle progression and maintenance of stem cell properties.

Author Summary Stem cell self-renewal has two essential elements, cell division and at least of one of the daughter cells retaining stem cell properties, so-called stemness. The interconnections between cell cycle and cell fate specification have been explored in embryonic stem cells. However, less is known about how cell cycle affects the cell fate decision in tissue stem cells. In this study, we explore the function of particular mitotic factors Ccny and Ccnyl1 in regulating the dividing tissue stem cells. The development of the mammary gland occurs mostly in postnatal pubertal stage. At the time, the robustly dividing stem/progenitor cells reside at the forefront of the mammary epithelium extension, underlining the mammary gland as a good model to study the interconnection of cell cycle and tissue stem cells. In this study, we show that in dividing mammary stem/progenitor cells, Ccny and Ccnyl1 enhance Wnt signaling activities in mitosis. The signaling enhancement in this time window is essential for the stem/progenitor cell property maintenance during division. Deletion of Ccnys results in diminishing their competitiveness and developmental potential.