Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain; Instituto Universitario de Investigación Mixto Agroalimentario de Aragón (IA2) UNIZAR-CITA, Zaragoza, Spain; Instituto de Investigación Sanitaria de Aragón (IIS-Aragón), Zaragoza, Spain.
Instituto Universitario de Investigación Mixto Agroalimentario de Aragón (IA2) UNIZAR-CITA, Zaragoza, Spain; Instituto de Investigación Sanitaria de Aragón (IIS-Aragón), Zaragoza, Spain; Centro de Encefalopatías y Enfermedades Transmisibles Emergentes (CEETE), Facultad de Veterinaria, Universidad de Zaragoza, Zaragoza, Spain.
Res Vet Sci. 2024 Nov;180:105423. doi: 10.1016/j.rvsc.2024.105423. Epub 2024 Sep 25.
In neurodegenerative diseases, including prion diseases, cellular models arise as useful tools to study the pathogenic mechanisms occurring in these diseases and to assess the efficacy of potential therapeutic compounds. In the present study, a RNA-sequencing analysis of bone marrow-derived ovine mesenchymal stem cells (oBM-MSCs) exposed to scrapie brain homogenate was performed to try to unravel genes and pathways potentially involved in prion diseases and MSC response mechanisms to prions. The oBM-MSCs were cultured in three different conditions (inoculated with brain homogenate of scrapie-infected sheep, with brain homogenate of healthy sheep and in standard growth conditions without inoculum) that were analysed at two exposure times: 2 and 4 days post-inoculation (dpi). Differentially expressed genes (DEGs) in scrapie-treated oBM-MSCs were found in the two exposure times finding the higher number at 2 dpi, which coincided with the inoculum removal time. Pathways enriched in DEGs were related to biological functions involved in prion toxicity and MSC response to the inflammatory environment of scrapie brain homogenate. Moreover, RNA-sequencing analysis was validated amplifying by RT-qPCR a set of 11 DEGs with functions related with prion propagation and its associated toxicity. Seven of these genes displayed significant expression changes in scrapie-treated cells. These results contribute to the knowledge of the molecular mechanisms behind the early toxicity observed in these cells after prion exposure and to elucidate the response of MSCs to neuroinflammation.
在神经退行性疾病中，包括朊病毒病，细胞模型作为研究这些疾病中发生的致病机制和评估潜在治疗化合物疗效的有用工具出现。在本研究中，对暴露于传染性海绵状脑病脑匀浆的骨髓来源的绵羊间充质干细胞（oBM-MSCs）进行了 RNA 测序分析，试图揭示可能与朊病毒病和 MSC 对朊病毒反应机制相关的基因和途径。oBM-MSCs 在三种不同条件下（接种传染性绵羊海绵状脑病脑匀浆、接种健康绵羊脑匀浆和标准生长条件下无接种物）培养，并在两种暴露时间（接种后 2 和 4 天）进行分析。在 2 dpi 时发现朊病毒处理的 oBM-MSCs 中的差异表达基因（DEGs）数量更高，这与接种物去除时间一致。DEGs 富集的途径与涉及朊病毒毒性和 MSC 对传染性海绵状脑病脑匀浆炎症环境的反应的生物学功能有关。此外，通过 RT-qPCR 扩增了一组与朊病毒传播及其相关毒性相关功能的 11 个 DEGs，对 RNA 测序分析进行了验证。这 7 个基因在朊病毒处理的细胞中显示出显著的表达变化。这些结果有助于了解这些细胞在暴露于朊病毒后早期观察到的毒性背后的分子机制，并阐明 MSC 对神经炎症的反应。
