Huang Jinbo, Teixeira Ana Palma, Gao Ting, Xue Shuai, Xie Mingqi, Fussenegger Martin
Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China.
Nat Commun. 2025 Feb 27;16(1):2028. doi: 10.1038/s41467-025-57275-x.
Current small-molecule-regulated synthetic gene switches face clinical limitations such as cytotoxicity, long-term side-effects and metabolic disturbances. Here, we describe an advanced synthetic platform inducible by risk-free input medication (ASPIRIN), which is activated by acetylsalicylic acid (ASA/aspirin), a multifunctional drug with pain-relieving, anti-inflammatory, and cardiovascular benefits. To construct ASPIRIN, we repurpose plant salicylic acid receptors NPR1 and NPR4. Through domain truncations and high-throughput mutant library screening, we enhance their ASA sensitivity. Optimized NPR1 fused with a membrane-tethering myristoylation signal (Myr-NPR1) forms a complex with NPR4, which is fused with a DNA binding domain (VanR) and a transactivation domain (VP16). ASA induces dissociation of the Myr-NPR1/NPR4-VanR-VP16 complex, allowing nuclear translocation of NPR4-VanR-VP16 to activate VanR-operator-controlled gene expression. In male diabetic mice implanted with microencapsulated ASPIRIN-engineered cells, ASA regulates insulin expression, restores normoglycemia, alleviates pain and reduces biomarkers of diabetic neuropathy and inflammation. We envision this system will pave the way for aspirin-based combination gene therapies.
当前小分子调控的合成基因开关面临细胞毒性、长期副作用和代谢紊乱等临床局限性。在此，我们描述了一种由无风险输入药物（阿司匹林）诱导的先进合成平台，该平台由乙酰水杨酸（ASA/阿司匹林）激活，乙酰水杨酸是一种具有止痛、抗炎和心血管益处的多功能药物。为构建阿司匹林诱导的合成平台（ASPIRIN），我们重新利用了植物水杨酸受体NPR1和NPR4。通过结构域截短和高通量突变文库筛选，我们提高了它们对ASA的敏感性。优化后的NPR1与膜锚定肉豆蔻酰化信号（Myr-NPR1）融合，与NPR4形成复合物，NPR4与DNA结合结构域（VanR）和反式激活结构域（VP16）融合。ASA诱导Myr-NPR1/NPR4-VanR-VP16复合物解离，使NPR4-VanR-VP16发生核转位，从而激活VanR操纵子控制的基因表达。在植入微囊化ASPIRIN工程细胞的雄性糖尿病小鼠中，ASA调节胰岛素表达，恢复正常血糖水平，减轻疼痛，并降低糖尿病神经病变和炎症的生物标志物水平。我们设想该系统将为基于阿司匹林的联合基因疗法铺平道路。
