Mutations in the glucocerebrosidase gene (GBA) cause Gaucher disease (GD), the lysosomal storage disorder (LSD), and are the most common genetic risk factor of Parkinson's disease (PD). Lysosome functionality plays a critical role for secretion of extracellular vesicles (EVs) and their content. Here we compared EVs from the blood plasma of 8 GD patients and 8 controls in terms of amounts, size distribution, and composition of their protein cargo. EVs were isolated via sequential centrifugation and characterized by сryo-electron microscopy (cryo-EM), nanoparticle tracking analysis (NTA), and dynamic light scattering (DLS). The presence of exosomal markers HSP70 and tetrasponins were analyzed by Western blot and flow cytometry. Protein profiling was performed by mass-spectrometry (shotgun analysis).

Here, for the first time we reported an increased size and altered morphology in exosomes derived from blood plasma of GD patients. An increased size of plasma exosomes from GD patients compared to controls was demonstrated by cryo-EM and DLS (р<0.0001, p < 0.001, respectively) and confirmed by mode size detected by NTA (p < 0.02). Cryo-EM demonstrated an increased number of double and multilayer vesicles in plasma EVs from GD patients. We found that the EVs were enriched with the surface exosomal markers (CD9, СD63, CD81) and an exosome-associated protein HSP70 in case of the patients with the disease. Proteomic profiling of exosomal proteins did not reveal any proteins associated with PD pathogenesis. Thus, we showed that lysosomal dysfunction in GD patients lead to a striking alteration of plasma exosomes in size and morphology.

葡糖脑苷脂酶基因（GBA）的突变会引起高雪氏病（GD），溶酶体贮积病（LSD），并且是帕金森氏病（PD）的最常见遗传危险因素。溶酶体功能对于细胞外囊泡（EV）及其内容的分泌起关键作用。在这里，我们比较了8例GD患者和8例对照的血浆中的电动汽车的数量，大小分布和蛋白货物的组成。电动汽车通过顺序离心分离，并通过电子显微镜（cryo-EM），纳米粒子跟踪分析（NTA）和动态光散射（DLS）进行表征。通过蛋白质印迹和流式细胞术分析了外泌体标志物HSP70和四联素的存在。通过质谱分析（shot弹枪分析）进行蛋白质谱分析。

在这里，我们首次报道了GD患者血浆中外泌体的大小增加和形态改变。冷冻EM和DLS证实了GD患者血浆外泌体的大小与对照组相比有所增加（分别р<0.0001，p <0.001），并由NTA检测到了模式大小（p <0.02）。Cryo-EM证实了GD患者血浆电动汽车中双层和多层囊泡的数量增加。我们发现，在患有该病的患者中，EV富含表面外泌体标志物（CD9，СD63，CD81）和与外泌体相关的蛋白HSP70。蛋白质组学的外泌体蛋白分析没有显示与PD发病机理相关的任何蛋白。因此，我们表明GD患者的溶酶体功能障碍导致血浆外泌体的大小和形态发生显着改变。