Impact of Hydatid Disease Antigens on the Transcriptomic Profile of Mouse Bone Marrow-Derived Mesenchymal Stem Cells
This study explores the transcriptomic alterations in mouse bone marrow-derived mesenchymal stem cells (BMSCs) upon exposure to antigens from Echinococcus granulosus, the causative agent of hydatid disease. Utilizing high-throughput sequencing, we analyzed the mRNA expression profiles of BMSCs treated with excretory-secretory products (ESPs), hydatid cyst fluid (HCF), and particles from the laminated layer (pLL). Our objective was to identify differentially expressed genes (DEGs) and investigate their roles in immune response modulation and cell behavior, including cell cycle and migration. The findings aim to enhance understanding of the interactions between parasitic antigens and host stem cells, potentially revealing novel therapeutic targets for hydatid disease. Overall design: 1. **Antigen Preparation:** ESPs, HCF, and pLL antigens are extracted from Echinococcus granulosus cysts obtained from sheep livers. 2. **BMSC Culture and Treatment:** BMSCs are isolated from C57BL/6 mice, cultured, and then treated with 2 µg/ml of each antigen type for 48 hours. 3. **RNA Extraction and Sequencing:** Total RNA is extracted from treated and control BMSCs, followed by library construction for Illumina NovaSeq sequencing. 4. **Data Analysis:** Sequencing data are analyzed to identify DEGs using Deseq2. Further bioinformatics analyses, including GO and KEGG enrichment, are performed to elucidate the biological implications of these DEGs. 5. **Validation:** Selected DEGs are validated through quantitative PCR and Western blot analysis. 6. **Functional Assays:** The effects of antigens on BMSCs are assessed, including cell cycle, apoptosis, and migration assays.