Differential Proteomic Analysis of Syncytiotrophoblast Extracellular Vesicles from Early-Onset Severe Preeclampsia, using 8-Plex iTRAQ Labeling Coupled with 2D Nano LC-MS/MS

Abstract

Aims: Previous studies have revealed that the increased shedding of syncytiotrophoblast extracellular vesicles (STBM) may lead to preeclampsia (PE). We aimed to identify the proteins carried by STBM and their potential pathological roles in early-onset severe PE. Methods: In this study, we performed a differential proteomic analysis of STBM from early-onset severe PE patients, using iTRAQ isobaric tags and 2D nano LC-MS/MS. STBM were generated by the in vitro explant culture method, and then verified by electron microscopy and western blot analysis. Results: A total of 18 533 unique peptides and 3 317 proteins were identified, 3 292 proteins were quantified. We identified 194 differentially expressed proteins in STBM from early-onset severe PE patients, 122 proteins were up-regulated and 72 proteins were down-regulated. Further bioinformatics analysis revealed that mitochondrion, transmembrane transport and transmembrane transporter activity were the most abundant categories in gene ontology (GO) annotation. Glycolysis/ gluconeogenesis, citrate cycle, fatty acid elongation, steroid hormone biosynthesis and oxidative phosphorylation were the five significantly represented pathways. Four differentially expressed proteins (siglec-6, calnexin, CD63 and S100-A8) related to inflammation, coagulation or immunoregulation were independently verified using western blot. Conclusions: The identification of key proteins carried by STBM may serve not only as a basis for better understanding and further exploring the etiology and pathogenesis of PE, but also as potential biomarkers and in providing targets for future therapy in PE, especially in early-onset severe PE(sPE).

AMPK Hyper-Activation Alters Fatty Acids Metabolism and Impairs Invasiveness of Trophoblasts in Preeclampsia

“…In conclusion, although the discrepancies in transcriptome and proteome between PE and normal placentas have been found previously [55,56], this study is the first to report PE placentas were associated with AMPK hyper-activation and an aberrant FA metabolomic profile. Persistent low oxygen conditions were required to induce 'PE-like' metabolic features in HTR8/SVneo cells.…”

Isolation of human trophoblastic extracellular vesicles and characterization of their cargo and antiviral activity

“…Consistent with this possibility, distinctive combinations of integrin subunits, such as α6β4 and αVβ5, were recently shown to target tumor EX metastasis to the lung and liver, respectively [80]. It is also possible the EV cargo is influenced by disease state, as recently suggested in the context of preeclampsia [81–83], as well as the stage of pregnancy and the methodology used for analysis. [84].…”

Cell-Derived Microparticles in Patients with Type 2 Diabetes Mellitus: a Systematic Review and Meta-Analysis

“…Microparticles (MPs), small membrane vesicles measuring 0.1~1.0 μm in diameter, are produced during the apoptosis and stimulation of various cell types, such as platelets, leukocytes, monocytes, erythrocytes, and endothelial cells [4][5][6][7]. Being overlooked for years, MPs are now considered as extracellular organelles with a variety of functions involving coagulation, inflammation, vascular function and transfer of biologic information [8][9][10][11].…”