Gong Xu, Liu Shan, Xia Bozhang, Wan Yichen, Zhang Shuyi, Zhang Baoyan, Wang Zehao, Chen Junge, Xiao Fei, Liang Xing-Jie, Yang Yun
Beijing Advanced Innovation Center for Biomedical Engineering, School of Medical Science and Engineering, Beihang University, Beijing, P. R. China.
Key Laboratory of Big Data-Based Precision Medicine, Ministry of Industry and Information Technology, Beihang University, Beijing, P. R. China.
Nat Commun. 2025 Feb 21;16(1):1862. doi: 10.1038/s41467-025-57153-6.
Genetically engineered commensal bacteria are promising living drugs, however, their therapeutic molecules are frequently confined to their colonization sites. Herein, we report an oral protein delivery technology utilizing an engineered bacterial type zero secretion system (T0SS) via outer membrane vesicles (OMVs). We find that OMVs produced in situ by Escherichia coli Nissle 1917 (EcN) can penetrate the intact gut epithelial barrier to enter the circulation and that epithelial transcytosis involves pinocytosis and dynamin-dependent pathways. EcN is engineered to endogenously load various enzymes into OMVs, and the secreted enzyme-loaded OMVs are able to stably catalyze diverse detoxification reactions against digestive fluid and even enter the circulation. Using hyperuricemic mice and uricase delivery as a demonstration, we demonstrate that the therapeutic efficacy of our engineered EcN with a modified T0SS outperforms that with a direct protein secretion apparatus. The enzyme-loaded OMVs also effectively detoxify human serum samples, highlighting the potential for the clinical treatment of metabolic disorders.
基因工程共生细菌是很有前景的活体药物，然而，它们的治疗分子常常局限于其定殖位点。在此，我们报告了一种口服蛋白质递送技术，该技术利用工程化细菌零分泌系统（T0SS）通过外膜囊泡（OMV）进行递送。我们发现，由大肠杆菌Nissle 1917（EcN）原位产生的OMV能够穿透完整的肠道上皮屏障进入循环系统，并且上皮细胞的转胞吞作用涉及胞饮作用和动力蛋白依赖性途径。对EcN进行工程改造，使其能够将各种酶内源性地装载到OMV中，分泌的装载酶的OMV能够稳定地催化针对消化液的各种解毒反应，甚至进入循环系统。以高尿酸血症小鼠和尿酸酶递送为例，我们证明，经过改造的T0SS的工程化EcN的治疗效果优于具有直接蛋白质分泌装置的EcN。装载酶的OMV也能有效解毒人血清样本，突出了其在代谢紊乱临床治疗中的潜力。
