Zhang Yue, Sun Wei, Wu Hao, Zhang Yi, Zhao Jianhong, Wu Shengbo, Yuan Lin, Chen Chen, Zheng Bin, Qiao Jianjun
Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, PR China.
Biomedical engineering, Academy of Medical Engineering and Translational Medicine, Tianjin University, Tianjin 300072, PR China.
Microbiol Res. 2025 Oct;299:128221. doi: 10.1016/j.micres.2025.128221. Epub 2025 May 26.
Engineered bacteria play an important role in colorectal disease. Lactococcus lactis (L. lactis) can inhibit Clostridioides difficile (C. difficile) by producing antimicrobial peptide nisin. However, its insufficient nisin production levels, lack of targeted in vivo release and weak colonization ability may limit its therapeutic efficacy against C. difficile infection (CDI). In this study, we engineered a strain of L. lactis with high adherence and therapeutic potential (Lla+) by expressing C. difficile adhesion protein Cwp8 on the surface of a L. lactis strain with high nisin yield (4019 to 4028 IU/ml). The adhesion effect of Lla+ was increased by 1.3-fold compared to the wild type L. lactis in the HT-29 cell model. We created the lipid membrane-coated L. lactis (LCL) by encapsulating Lla+ with the ROS-responsive lipid membrane. LCL exhibited targeted release of nisin payload in response to HO, lipopolysaccharide and C. difficile in vitro, as well as in various mouse models of intestinal inflammation in vivo. The therapeutic effect of LCL against CDI was substantial, reducing C. difficile survival by 60 % compared to the untreated control. In the treatment of recurrent CDI (rCDI), LCL outperformed the drug fidaxomicin, the first-line treatment for rCDI recommended by Infectious Diseases Society of America (IDSA), decreasing C. difficile survival by 48 % and lowering the levels of toxins TcdA and TcdB by 57.1 % and 65 %, respectively. These results suggested that the presented the encapsulation approach could serve as a delivery platform to target inflamed intestines and expand the application of probiotics as pharmaceuticals.
工程菌在结直肠疾病中发挥着重要作用。乳酸乳球菌（L. lactis）可通过产生抗菌肽乳链菌素来抑制艰难梭菌（C. difficile）。然而，其乳酸链球菌素产量不足、缺乏体内靶向释放能力以及定殖能力较弱，可能会限制其对艰难梭菌感染（CDI）的治疗效果。在本研究中，我们通过在高产乳酸链球菌素（4019至4028 IU/ml）的乳酸乳球菌菌株表面表达艰难梭菌黏附蛋白Cwp8，构建了一株具有高黏附性和治疗潜力的乳酸乳球菌（Lla+）。在HT - 29细胞模型中，Lla+的黏附效果比野生型乳酸乳球菌提高了1.3倍。我们通过用ROS响应性脂质膜包裹Lla+，制备了脂质膜包被的乳酸乳球菌（LCL）。LCL在体外以及在各种体内肠道炎症小鼠模型中，对HO、脂多糖和艰难梭菌均表现出靶向释放乳酸链球菌素。LCL对CDI的治疗效果显著，与未治疗的对照组相比，艰难梭菌存活率降低了60%。在复发性CDI（rCDI）的治疗中，LCL的表现优于美国传染病学会（IDSA）推荐的rCDI一线治疗药物非达霉素，艰难梭菌存活率降低了48%，毒素TcdA和TcdB的水平分别降低了57.1%和65%。这些结果表明，所提出的封装方法可作为一种靶向炎症肠道的递送平台，并扩大益生菌作为药物的应用。
