Marten Andrew D, Haslitt Douglas P, Martin Chad A, Karthikeyan Akshitha, Swanson Daniel H, Kalera Karishma, Johnson Ulysses G, Swarts Benjamin M, Conway Michael J
Foundational Sciences, Central Michigan University College of Medicine, Mount Pleasant, Michigan, United States of America.
Department of Chemistry and Biochemistry, Central Michigan University, Mount Pleasant, Michigan, United States of America.
PLoS Pathog. 2025 May 6;21(5):e1012795. doi: 10.1371/journal.ppat.1012795. eCollection 2025 May.
Trehalose is a non-reducing disaccharide that is the major sugar found in insect hemolymph fluid. Trehalose provides energy, and promotes growth, metamorphosis, stress recovery, chitin synthesis, and insect flight. Trehalase is the only enzyme responsible for the hydrolysis of trehalose, which makes it an attractive molecular target. Here we show that Aedes aegypti (Aag2) cells express trehalase and that they can grow on trehalose-containing cell culture media. Trehalase activity was confirmed by treating Aag2 cells with trehalase inhibitors, which inhibited conversion of trehalose to glucose and reduced cell proliferation. Cell entry of a fluorescent trehalose probe was dependent on trehalose concentration, suggesting that trehalose moves across the cell membrane via passive transport. Culturing Aag2 cells with trehalose-containing cell culture media led to significant changes in gene expression, intracellular lipids, and dengue virus replication and specific infectivity, and increased their susceptibility to trehalase inhibitors. These data describe an in vitro model that can be used to rapidly screen novel trehalase inhibitors and probes and underscores the importance of trehalose metabolism in Ae. aegypti physiology and transmission of a mosquito-borne virus.
海藻糖是一种非还原性二糖，是昆虫血淋巴液中发现的主要糖类。海藻糖提供能量，并促进生长、变态、应激恢复、几丁质合成和昆虫飞行。海藻糖酶是唯一负责水解海藻糖的酶，这使其成为一个有吸引力的分子靶点。在这里，我们表明埃及伊蚊（Aag2）细胞表达海藻糖酶，并且它们可以在含海藻糖的细胞培养基上生长。通过用海藻糖酶抑制剂处理Aag2细胞来确认海藻糖酶活性，该抑制剂抑制了海藻糖向葡萄糖的转化并减少了细胞增殖。荧光海藻糖探针的细胞摄取取决于海藻糖浓度，这表明海藻糖通过被动运输穿过细胞膜。用含海藻糖的细胞培养基培养Aag2细胞导致基因表达、细胞内脂质、登革病毒复制和特定感染性发生显著变化，并增加了它们对海藻糖酶抑制剂的敏感性。这些数据描述了一种体外模型，可用于快速筛选新型海藻糖酶抑制剂和探针，并强调了海藻糖代谢在埃及伊蚊生理学和蚊媒病毒传播中的重要性。
