Abnormal Endocytosis Resulting from Imbalanced Proteomic Doses in Human Aneuploid Embryos

To explore transcriptome differences between diploid and aneuploidy embryos, identify non-invasive screening targets for aneuploidy embryos, and establish a theoretical basis for a pathogenic model. RNA sequencing compared transcriptomes of diploid and aneuploid blastocysts, with GO and pathway analysis on differentially expressed genes. Fluorescent probes assessed clathrin-dependent and independent endocytosis in both cell types, while fluorescence quantitative PCR validated endocytosis abnormalities in early aneuploidy development. Additionally, free amino acid content was compared to show how aneuploidy genome imbalance affects endocytosis via changes in osmotic pressure. RNA sequencing revealed 9731 differentially expressed genes between normal diploids and aneuploidys, with 6134 up-regulated and 3597 down-regulated. KEGG analysis indicated these genes are mainly involved in endocytosis-related pathways. Six genes (PSD3, ARAP2, SMAP2, CBLC, AGAP1, SH3GLB1) showed significant differences (P < 0.05) in expression between diploid and aneuploidy groups. Molecular probe analysis and Fluorescence quantitative PCR resultsrevealed reduced clathrin-dependent endocytosis and increased clathrin-independent endocytosis in aneuploidy embryos compared to normal diploids(P < 0.05). Additionally, aneuploid embryos showed higher relative abundance of 14 free amino acids, particularly methionine. The study concludes that early transcriptome differences in aneuploid embryos are centered on endocytosis. Normal diploid embryos primarily use clathrin-dependent pathways, whereas aneuploid embryos favor clathrin-independent pathways. The endocytosis abnormalities in aneuploid cells are due to changes in intracellular osmotic pressure. This study provides a potential target for non-invasive detection of aneuploid embryos and lays a theoretical foundation for the establishment of a pathogenicity prediction model for aneuploid embryos.