Murugesan Saravanakumar, Saravanakumar Lakshmi, Sadayappan Sakthivel, Kannappan Ramaswamy, Sinkey Rachel G, Tubinis Michelle D, Tita Alan N, Jilling Tamas, Berkowitz Dan E
Division of Molecular and Translational Biomedicine, Department of Anesthesiology and Perioperative Medicine, School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, United States.
Department of Cellular and Molecular Medicine, Sarver Heart Center, University of Arizona College of Medicine, Tucson, Arizona, United States.
Am J Physiol Cell Physiol. 2025 Jan 1;328(1):C128-C138. doi: 10.1152/ajpcell.00409.2024. Epub 2024 Dec 9.
Women with severe preeclampsia (sPE) exhibit a heightened risk of postpartum cardiovascular disease compared with those with normotensive pregnancies (NTP). Although placental extracellular vesicles (EVs) play a crucial role in feto-maternal communication, their impact on cardiomyocytes, particularly in the context of sPE, remains unclear. This study investigated the effect of sPE-associated placental EVs (sPE-Plex EVs) on cardiomyocyte calcium dynamics. We hypothesized that sPE-Plex EV mediates cardiomyocyte dysfunction by disrupting calcium signaling. EVs were isolated from plasma and placental explant culture (Plex) using precipitation methods and confirmed as Plex EVs by placental alkaline phosphatase (PLAP) activity and electron microscopy. Moreover, confocal microscopy confirmed the uptake of plasma EVs in human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and Plex EVs by human AC-16 cardiomyocyte (hAC-16CM) cells. hiPSC-CM cells treated with sPE-EVs and hAC-16CM cells treated with sPE-Plex EVs exhibited significantly lower levels of stromal interaction molecule 1 (STIM1) and phospholamban (PLN) proteins compared with those treated with normotensive controls EVs, as confirmed by Western blot analysis. Treatment with sPE-Plex EVs also resulted in the downregulation of STIM1 and PLN proteins in murine cardiomyocyte (mCM) cells compared with treatment with NTP-Plex EVs. Our findings suggest that both plasma EVs and Plex EVs from sPE may alter calcium signaling in cardiac cells by downregulating calcium sensor proteins (STIM1 and PLN). Therefore, plasma EVs and Plex EVs from sPE pregnancies have adverse effects by altering calcium dynamics in hiPSC-CM, hAC-16CM, and mCM compared with normotensive control and potential impairment of cardiomyocyte function ex vivo. This study unveils a novel link between the placenta and PE-linked heart dysfunction. We isolated and characterized placental EVs from pregnancies with sPE and normotensive controls. These plasma sPE-EVs, and sPE-Plex EVs disrupt calcium signaling in heart cells, potentially via reduced STIM1 and PLN proteins. This suggests both plasma sPE-EVs and sPE-Plex EVs cargo drive these disruptive effects. Identifying these cargo molecules (miRNAs or proteins) holds promise for new PE therapies targeting cardiac dysfunction.
与血压正常的孕妇（NTP）相比，重度子痫前期（sPE）女性产后患心血管疾病的风险更高。尽管胎盘细胞外囊泡（EVs）在母婴交流中起着关键作用，但其对心肌细胞的影响，尤其是在sPE背景下，仍不清楚。本研究调查了sPE相关胎盘EVs（sPE-Plex EVs）对心肌细胞钙动力学的影响。我们假设sPE-Plex EVs通过破坏钙信号传导介导心肌细胞功能障碍。使用沉淀方法从血浆和胎盘外植体培养物（Plex）中分离出EVs，并通过胎盘碱性磷酸酶（PLAP）活性和电子显微镜确认为Plex EVs。此外，共聚焦显微镜证实了人诱导多能干细胞衍生的心肌细胞（hiPSC-CMs）对血浆EVs的摄取以及人AC-16心肌细胞（hAC-16CM）对Plex EVs的摄取。与用血压正常对照的EVs处理的细胞相比，用sPE-EVs处理的hiPSC-CM细胞和用sPE-Plex EVs处理的hAC-16CM细胞中基质相互作用分子1（STIM1）和受磷蛋白（PLN）蛋白水平显著降低，蛋白质印迹分析证实了这一点。与用NTP-Plex EVs处理相比，用sPE-Plex EVs处理也导致小鼠心肌细胞（mCM）中STIM1和PLN蛋白下调。我们的研究结果表明，来自sPE的血浆EVs和Plex EVs可能通过下调钙传感器蛋白（STIM1和PLN）改变心脏细胞中的钙信号传导。因此，与血压正常对照相比，来自sPE妊娠的血浆EVs和Plex EVs通过改变hiPSC-CM、hAC-16CM和mCM中的钙动力学产生不利影响，并可能在体外损害心肌细胞功能。本研究揭示了胎盘与PE相关心脏功能障碍之间的新联系。我们从患有sPE的妊娠和血压正常对照中分离并表征了胎盘EVs。这些血浆sPE-EVs和sPE-Plex EVs可能通过降低STIM1和PLN蛋白破坏心脏细胞中的钙信号传导。这表明血浆sPE-EVs和sPE-Plex EVs的货物都驱动了这些破坏作用。鉴定这些货物分子（miRNA或蛋白质）有望为针对心脏功能障碍的新PE疗法带来希望。
