Mundell Jordan W, Brier Matthew I, Orloff Everest, Stanley Sarah A, Dordick Jonathan S
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
iScience. 2024 Feb 12;27(3):109186. doi: 10.1016/j.isci.2024.109186. eCollection 2024 Mar 15.
Magnetogenetics represents a method for remote control of cellular function. Previous work suggests that generation of reactive oxygen species (ROS) initiates downstream signaling. Herein, a chemical biology approach was used to elucidate further the mechanism of radio frequency-alternating magnetic field (RF-AMF) stimulation of a TRPV1-ferritin magnetogenetics platform that leads to Ca flux. RF-AMF stimulation of HEK293T cells expressing TRPV1-ferritin resulted in ∼30% and ∼140% increase in intra- and extracellular ROS levels, respectively. Mutations to specific cysteine residues in TRPV1 responsible for ROS sensitivity eliminated RF-AMF driven Ca-dependent transcription of secreted embryonic alkaline phosphatase (SEAP). Using a non-tethered (to TRPV1) ferritin also eliminated RF-AMF driven SEAP production, and using specific inhibitors, ROS-activated TRPV1 signaling involves protein kinase C, NADPH oxidase, and the endoplasmic reticulum. These results suggest ferritin-dependent ROS activation of TRPV1 plays a key role in the initiation of magnetogenetics, and provides relevance for potential applications in medicine and biotechnology.
磁遗传学是一种用于远程控制细胞功能的方法。先前的研究表明，活性氧（ROS）的产生会启动下游信号传导。在此，采用化学生物学方法进一步阐明射频交变磁场（RF-AMF）刺激TRPV1-铁蛋白磁遗传学平台导致钙通量的机制。RF-AMF刺激表达TRPV1-铁蛋白的HEK293T细胞分别导致细胞内和细胞外ROS水平增加约30%和约140%。TRPV1中负责ROS敏感性的特定半胱氨酸残基的突变消除了RF-AMF驱动的分泌型胚胎碱性磷酸酶（SEAP）的钙依赖性转录。使用非 tethered（与TRPV1相连）的铁蛋白也消除了RF-AMF驱动的SEAP产生，并且使用特异性抑制剂表明，ROS激活的TRPV1信号传导涉及蛋白激酶C、NADPH氧化酶和内质网。这些结果表明，铁蛋白依赖性的TRPV1的ROS激活在磁遗传学的启动中起关键作用，并为医学和生物技术中的潜在应用提供了相关性。
