Bravo-Miana Rocío Del Carmen, Arizaga-Echebarria Jone Karmele, Sabas-Ortega Valeria, Crespillo-Velasco Hirune, Prada Alvaro, Castillo-Triviño Tamara, Otaegui David
Neuroimmunology Group, Neuroscience Area, Biogipuzkoa Health Research Institute, 20014 San Sebastián, Spain.
Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas, Instituto de Salud Carlos III, 28029 Madrid, Spain.
Biomedicines. 2024 Oct 2;12(10):2245. doi: 10.3390/biomedicines12102245.
This study aimed to unravel the single tetraspanin pattern of extracellular vesicles (EVs), L1CAM and GLAST EV levels as diagnostic biomarkers to stratify people with multiple sclerosis (pwMS), specifically relapsing-remitting (RRMS) and primary progressive (PPMS). The ExoView platform was used to directly track single EVs using a clinically feasible volume of cerebrospinal fluid (CSF) and serum samples. This technology allowed us to examine the patterns of classical tetraspanin and quantify the levels of L1CAM and GLAST proteins, commonly used to immunoisolate putative neuron- and astrocyte-derived EVs. The tetraspanin EV pattern does not allow us to differentiate RRMS, PPMS and non-MS donors neither in CSF nor serum, but this was associated with the type of biofluid. L1CAM and GLAST EVs showed a very low presence of tetraspanin proteins. Additionally, a significant decrease in the particle count of L1CAM EVs was detected in L1CAM-captured spots, and L1CAM and GLAST EVs decreased in GLAST-captured spots in the CSF from PPMS subjects compared to RRMS. Interestingly, only GLAST EVs exhibited a lower quantity in the CSF from PPMS compared to both MS and non-MS samples. Finally, GLAST EVs demonstrated a medium negative and significative correlation with GFAP levels-a biomarker of MS progression, astrocyte damage and neurodegenerative processes. ExoView technology could track neural EV biomarkers and be potentially useful in the diagnostic evaluation and follow-up of pwMS. GLAST EVs might provide insights into the etiology of PPMS and could offer small windows to elucidate the molecular mechanisms behind its clinical presentation.
本研究旨在揭示细胞外囊泡（EVs）的单一四跨膜蛋白模式、L1CAM和GLAST EV水平，将其作为诊断生物标志物，对多发性硬化症患者（pwMS），特别是复发缓解型（RRMS）和原发进展型（PPMS）进行分层。ExoView平台用于使用临床可行体积的脑脊液（CSF）和血清样本直接追踪单个EVs。这项技术使我们能够检查经典四跨膜蛋白的模式，并量化L1CAM和GLAST蛋白的水平，这些蛋白通常用于免疫分离假定的神经元和星形胶质细胞衍生的EVs。四跨膜蛋白EV模式在CSF和血清中均无法使我们区分RRMS、PPMS和非MS供体，但这与生物流体的类型有关。L1CAM和GLAST EVs显示四跨膜蛋白的存在非常低。此外，在L1CAM捕获的斑点中检测到L1CAM EVs的颗粒计数显著减少，与RRMS相比，PPMS受试者CSF中GLAST捕获斑点中的L1CAM和GLAST EVs减少。有趣的是，与MS和非MS样本相比，仅GLAST EVs在PPMS的CSF中含量较低。最后，GLAST EVs与GFAP水平呈中度负相关且具有显著意义，GFAP是MS进展、星形胶质细胞损伤和神经退行性过程的生物标志物。ExoView技术可以追踪神经EV生物标志物，可能对pwMS的诊断评估和随访有用。GLAST EVs可能为PPMS的病因提供见解，并可能为阐明其临床表现背后的分子机制提供小窗口。
