Generation of Human Expanded Pluripotent Stem Cell-Derived Kidney Organoids with Patterned Nephron by Regulating Hippo-YAP Pathway

Background: Numerous cultivation protocols for kidney organoids, differentiation efficiency remains generally low. This study aims to investigate the influence of the Hippo-YAP pathway on induced differentiation of kidney organoids and elucidate its molecular mechanisms, thereby providing efficient induction strategies for kidney organoid construction. Methods: We used CRISPR-Cas9 technology for a reporter gene cell line was constructed based on human Expanded Pluripotent Stem Cells (hEPSC). Besides, an inducible YAP overexpression cell line and a NPHP1 knock-out cell line were constructed on this basis. The differentiation of kidney organoids under genetic and chemical schemes were compared. Results: We have developed a methodology to generate kidney organoids with pattern nerphron, differentiated from SIX2-mCherry knock-in reporter hEPSCs, by regulating Hippo-YAP signaling pathway. We find that inhibiting the Hippo-YAP pathway during early differentiation stages effectively increased the number of nephron progenitor cells, resulting in more pattern nerphron. Conclusion: Regulating the Hippo-YAP pathway during early stages of kidney organoid differentiation facilitates their maturation, offering effective induction strategies to enhance the efficiency of kidney organoid differentiation.The result for regualting YAP expressionSchematic showing regulating Hippo-YAP pathway for kidney organoids with patterned nephron