Damage-Associated molecular pattern markers HMGB1 and cell-Free fetal telomere fragments in oxidative-Stressed amnion epithelial cell-Derived exosomes

• Published in: Journal of reproductive immunology 2017-09, Vol.123, p.3-11
• Main Authors: Sheller-Miller, Samantha, Urrabaz-Garza, Rheanna, Saade, George, Menon, Ramkumar
• Format: Article
• Language: English
• Subjects: Alarmins - metabolism; Amniochorion; Amnion - physiology; Cells, Cultured; Cellular Senescence; Cigarette Smoking - adverse effects; Culture Media, Conditioned - adverse effects; DAMPs; Epithelial Cells - pathology; Exosomes - metabolism; Exosomes - pathology; Female; Fetal signals; HMGB1 Protein - metabolism; Humans; Inflammation; Inflammation - genetics; Inflammation - metabolism; Labor, Obstetric; Microvesicles; Oxidative Stress; Parturition; Pregnancy; Primary Cell Culture; Telomere - metabolism
• ISSN: 0165-0378; 1872-7603
• DOI: 10.1016/j.jri.2017.08.003

•Oxidative stress induces primary amnion cells to undergo cellular senescence.•Senescence results in damage-associated molecular pattern (DAMPs) release.•Senescence causes nuclear to cytosolic migration of DAMPs HMGB1/telomere fragments.•Cytoplasmic HMGB1 and telomere fragments are packaged inside exosomes.•HMGB1 and telomere fragments represent sterile inflammation propagated via exosomes. Term labor in humans is associated with increased oxidative stress (OS) −induced senescence and damages to amnion epithelial cells (AECs). Senescent fetal cells release alarmin high-mobility group box 1 (HMGB1) and cell-free fetal telomere fragments (cffTF) which can be carried by exosomes to other uterine tissues to produce parturition-associated inflammatory changes. This study characterized AEC-derived exosomes under normal and OS conditions and their packaging of HMGB1 and cffTF. Primary AECs were treated with either standard media or oxidative stress-induced media (exposure to cigarette smoke extract for 48h). Senescence was determined, and exosomes were isolated and characterized. To colocalize HMGB1 and cffTF in amnion exosomes, immunofluorescent staining and in situ hybridization were performed, followed by confocal microscopy. Next generation sequencing (NGS) determined exosomal cffTF and other cell-free amnion cell DNA specificity. Regardless of condition, primary AECs produce exosomes with a classic size, shape, and markers. OS and senescence caused the translocation of HMGB1 and cffTF from AECs’ nuclei to cytoplasm compared to untreated cells, which was inhibited by antioxidant N-acetyl cysteine (NAC). Linescans confirmed colocalization of HMGB1 and cffTF in exosomes were higher in the cytoplasm after CSE treatment compared to untreated AECs. NGS determined that besides cffTF, AEC exosomes also carry genomic and mitochondrial DNA, regardless of growth conditions. Sterile inflammatory markers HMGB1 and cffTF from senescent fetal cells are packaged inside exosomes. We postulate that this exosomal cargo can act as a fetal signal at term and can cause labor-associated changes in neighboring tissues.