Yoshida Yuya, Tanihara Tomohito, Hamasaki Keika, Tsurusaki Fumiaki, Fukuda Taiki, Adachi Satoka, Terada Yuma, Otsuki Kaita, Nishikawa Naoki, Fukuoka Kohei, Tsukamoto Ryotaro, Hamamura Kengo, Oyama Kosuke, Tsuruta Akito, Mayanagi Kouta, Koyanagi Satoru, Ohdo Shigehiro, Matsunaga Naoya
Department of Clinical Pharmacokinetics, Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan.
Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan.
Theranostics. 2025 Jan 1;15(2):340-361. doi: 10.7150/thno.100748. eCollection 2025.
Macrophage phagocytosis plays a role in cancer immunotherapy. The phagocytic activity of macrophages, regulated by circadian clock genes, shows time-dependent variation. Intervening in the circadian clock machinery of macrophages is a potentially novel approach to cancer immunotherapy; however, data on this approach are scarce. Microcurrent stimulation (MCS) promotes inflammation, proliferation, and remodeling, suggesting its potential to modulate macrophage function; however, its application has been limited. In this study, we investigated the impact of MCS on macrophage phagocytosis of cancer cells using mouse/human macrophage cell lines and various mouse/human cancer cell lines. Cells and mice received 300 µA, 400 Hz bidirectional pulsed MCS. Gene expression, protein expression, and phagocytosis activity were assessed in intraperitoneal macrophages collected from mice, as well as in RAW264.7, and THP-1 cells. Flow cytometry, population, phagocytosis activity, RNA-seq, and immunohistochemistry analyses were performed. Noninvasive MCS prevented time-dependent reduction in macrophage phagocytosis of cancer cells by modulating the circadian clock genes. MCS also enhanced phagocytosis in mouse RAW264.7 and human THP-1 cells across various cancer types by promoting actin polymerization; similar effects were observed in mice. This enhancement occurred in abdominal macrophages of both sexes and was mediated by changes in clock gene expression. Specifically, suppressing the clock gene nullified the effects of MCS. Moreover, although macrophage phagocytosis typically declined during the dark period, MCS during the light period prevented this reduction. MCS also increased phagocytosis of peritoneally implanted cancer cells (4T1, ID8, and Hepa1-6) in mice, significantly reducing tumor engraftment and growth, and ultimately improving prognosis. The findings of this study suggest that targeting macrophage circadian mechanisms via MCS could enhance cancer immunity, offering new avenues for cancer immunotherapy.
巨噬细胞吞噬作用在癌症免疫治疗中发挥作用。巨噬细胞的吞噬活性受生物钟基因调控，呈现出时间依赖性变化。干预巨噬细胞的生物钟机制是一种潜在的新型癌症免疫治疗方法；然而，关于这种方法的数据很少。微电流刺激（MCS）可促进炎症、增殖和重塑，表明其具有调节巨噬细胞功能的潜力；然而，其应用一直有限。在本研究中，我们使用小鼠/人类巨噬细胞系和各种小鼠/人类癌细胞系，研究了MCS对巨噬细胞吞噬癌细胞的影响。细胞和小鼠接受300 µA、400 Hz的双向脉冲MCS。对从小鼠收集的腹腔巨噬细胞以及RAW264.7和THP-1细胞进行基因表达、蛋白质表达和吞噬活性评估。进行了流式细胞术、群体、吞噬活性、RNA测序和免疫组织化学分析。非侵入性MCS通过调节生物钟基因，防止巨噬细胞对癌细胞的吞噬作用出现时间依赖性降低。MCS还通过促进肌动蛋白聚合，增强了小鼠RAW264.7和人类THP-1细胞对各种癌症类型的吞噬作用；在小鼠中也观察到了类似的效果。这种增强作用在两性的腹腔巨噬细胞中均有发生，并且是由生物钟基因表达的变化介导的。具体而言，抑制生物钟基因可消除MCS的作用。此外，尽管巨噬细胞吞噬作用通常在黑暗期下降，但光照期的MCS可防止这种下降。MCS还增加了小鼠腹膜植入癌细胞（4T1、ID8和Hepa1-6）的吞噬作用，显著减少肿瘤植入和生长，并最终改善预后。本研究结果表明，通过MCS靶向巨噬细胞生物钟机制可增强癌症免疫力，为癌症免疫治疗提供新途径。
