RESUMEN
BACKGROUND: Prior research has shown that granulin precursor (GRN, also termed PGRN) is closely linked to aphasia. However, there has been little research on the mechanism of action of GRN in post-stroke aphasia (PSA). METHODS: In this study, RT-qPCR was used to identify variations in gene expression, while RNA sequencing (RNA-seq) was utilized to acquire transcriptional profiles. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were employed for bioinformatics analysis. RESULTS: GRN was considerably more active in PSA subjects. After silencing the GRN, 197 transcripts had differential expression, and 237 alternative splicing events (ASEs) were substantially affected. The analysis of differentially expressed genes (DEGs) using GO and KEGG approaches showed that these genes have various molecular functions and are significantly enriched in metabolic signaling pathways. Regarding Alternative Splicing (AS), the GO and KEGG analyses revealed numerous functional genes involved in transcription and metabolism. CONCLUSIONS: The knockdown of GRN has been shown to be associated with alterations in transcription, metabolism, and ASEs, potentially impacting transcriptional and metabolic pathways through its involvement in AS. Furthermore, GRN knockdown is associated with nervous system disease-related gene transcription and AS processes, as well as its involvement in G protein-coupled receptor (GPCR) and wingless/integrated (Wnt) signaling pathways, which impact the initiation and resolution of PSA.
RESUMEN
Objective:To document the expression of aphasia-related progranulin gene (GRN) in mononuclear cells in the peripheral blood (PBMC) of patients with post-stroke aphasia (PSA).Methods:PC12 cells at the logarithmic-growth stage were cultured and divided into a non-specific interference group (the gene control group) and a specific interference group (the gene silencing group) when the cell density reached 30 to 50%. After the expression of GRN was knocked down in the cells, the occurrence of variable splicing events was analyzed using high-throughput transcriptome sequencing (RNA-seq). Meanwhile, 10 PSA patients were selected into a patient group and 10 healthy counterparts were chosen as a control group. Blood was collected from both groups and real-time fluorescence quantitative polymerase chain reactions (RT-qPCR) were employed to determine any changes in GRN mRNA expression and the occurrence of variable splicing events in the nuclear factor related to kappa-B-binding protein (NFRKB) in their PBMCs. The patient group received conventional speech therapy, and immediately after their first and second blood collections their speech functioning was assessed using the Chinese Aphasia Battery (ABC). Pearson correlation coefficients were then computed relating the GRN expression and ABC scores.Results:After knocking down GRN in the PC12 cells, the expression of GRN in the gene knockdown group was significantly different from that in the control group. There were 237 genes with significant differences in variable splicing between the two samples. The number of genes with variable splicing events at the 5′ end was the largest. There were also significant differences between the groups in the average occurrence of NFRKB variable splicing events. And significant diffe-rences were observed in the mRNA expression of GRN between the two blood collections from the patient group, as well as between the first collection from the patient group and the controls. The average oral expression score of the PSA patients improved significantly, particularly the retelling score. The changes in the GRN expression level were positively correlated with the recovery of oral expression ability.Conclusion:GRN can promote the recovery of speech function in PSA patients by regulating the variable splicing of NFRKB.
