Systemic proteomics and miRNA profile analysis of exosomes derived from human pluripotent stem cells

Abstract

Background Increasing studies have reported the therapeutic effect of mesenchymal stem cell (MSC)-derived exosomes by which protein and miRNA are clearly characterized. However, the proteomics and miRNA profiles of exosomes derived from human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs) remain unclear. Methods In this study, we isolated exosomes from hESCs, hiPSCs, and human umbilical cord mesenchymal stem cells (hUC-MSCs) via classic ultracentrifugation and a 0.22-μm filter, followed by the conservative identification. Tandem mass tag labeling and label-free relative peptide quantification together defined their proteomics. High-throughput sequencing was performed to determine miRNA profiles. Then, we conducted a bioinformatics analysis to identify the dominant biological processes and pathways modulated by exosome cargos. Finally, the western blot and RT-qPCR were performed to detect the actual loads of proteins and miRNAs in three types of exosomes. Results Based on our study, the cargos from three types of exosomes contribute to sophisticated biological processes. In comparison, hESC exosomes (hESC-Exos) were superior in regulating development, metabolism, and anti-aging, and hiPSC exosomes (hiPSC-Exos) had similar biological functions as hESC-Exos, whereas hUC-MSCs exosomes (hUC-MSC-Exos) contributed more to immune regulation. Conclusions The data presented in our study help define the protein and miRNA landscapes of three exosomes, predict their biological functions via systematic and comprehensive network analysis at the system level, and reveal their respective potential applications in different fields so as to optimize exosome selection in preclinical and clinical trials.

Induced pluripotent stem cell-derived extracellular vesicles promote wound repair in a diabetic mouse model via an anti-inflammatory immunomodulatory mechanism

“…This is particularly interesting in a diabetic wound healing model specifically, given the role of IGF1 in insulin regulation 49 . This result may also be supported by a recent study that profiled cargos within iPSC EVs, showing that many of these cargos are involved with modulating metabolism and aging, which IGF1 is also involved in 50, 51 .…”

Induced Pluripotent Stem Cell‐Derived Extracellular Vesicles Promote Wound Repair in a Diabetic Mouse Model via an Anti‐Inflammatory Immunomodulatory Mechanism

“…[45] This result may also be supported by a recent study that profiled cargos within iPSC EVs, showing that many of these cargos are involved with modulating metabolism and aging, which IGF1 is also involved in. [46,47] Given evidence that macrophage phenotypes are also related to cellular metabolism, it is possible that iPSC EVs may impart the observed "M2" transition through a similar pathway. [48][49][50] However, further studies into the mechanism behind these antiinflammatory phenomena are needed.…”

“…[ 45 ] This result may also be supported by a recent study that profiled cargos within iPSC EVs, showing that many of these cargos are involved with modulating metabolism and aging, which IGF1 is also involved in. [ 46,47 ]…”

Extracellular vesicle biogenesis of three‐dimensional human pluripotent stem cells in a novel Vertical‐Wheel bioreactor

“…The identified top pathways include ECM-receptor interaction, ECM organization, beta catenin-independent Wnt pathway, regulation of apoptosis and cell cycle-related G1/S transition pathway. The predicted miR regulators versus miRs in EVs identified in previous literature (Bi et al, 2022) include miR-222-3p, 423-3p, 484 and 92a-3p (Figure S8). The specific protein cargo related to Wnt pathways is shown in Table S6 and the negative regulation of apoptosis-related proteins is shown in Table S7.…”