Serebrinsky-Duek Kineret, Barra Maria, Danino Tal, Garrido Daniel
Department of Chemical and Bioprocess Engineering, School of Engineering, Pontificia Universidad Católica de Chile, Santiago, Chile.
Department of Biomedical Engineering, Columbia University, New York, New York, USA.
Microbiol Spectr. 2023 Mar 20;11(2):e0004923. doi: 10.1128/spectrum.00049-23.
Short-chain fatty acids (SCFA) such as propionate and butyrate are critical metabolites produced by the gut microbiota. Microbiome dysbiosis resulting in altered SCFA profiles is associated with certain diseases, including inflammatory bowel diseases (IBD), characterized by a reduction in butyrate concentration and active intestinal inflammation. There is an increasing interest in the use of engineered bacteria as diagnostic and therapeutic tools for gut diseases. In this study, we developed genetic circuits capable of sensing SCFA concentrations to build biosensors that express a response protein (superfolder green fluorescent protein [sfGFP]) in amounts inversely proportional to the SCFA concentration. We also built biotherapeutics expressing the cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF) using the same logic. The propionate biotherapeutic expressed larger amounts of mouse GM-CSF in the absence of propionate. The butyrate biotherapeutics presented the expected behavior only at the beginning of the kinetics and an accelerated response in the absence of butyrate. Overall, these genetic systems may function as complementary diagnostic tools for measuring SCFAs and as delivery vehicles for biotherapeutic molecules. Short-chain fatty acids are key molecules produced by the gut microbiome. Their concentrations are altered in certain diseases. Here, we created molecular biosensors that quantify the absence of propionate and butyrate, using logic "NOT" gates and bacterial promoters. Finally, we show that these genetic systems could be useful for the delivery of therapeutic molecules in the gut, in the absence of these acids.
短链脂肪酸（SCFA）如丙酸酯和丁酸酯是肠道微生物群产生的关键代谢产物。微生物群失调导致短链脂肪酸谱改变与某些疾病相关，包括炎症性肠病（IBD），其特征是丁酸酯浓度降低和肠道炎症活跃。人们越来越关注使用工程菌作为肠道疾病的诊断和治疗工具。在本研究中，我们开发了能够感知短链脂肪酸浓度的遗传回路，以构建生物传感器，该生物传感器表达的响应蛋白（超折叠绿色荧光蛋白[sfGFP]）的量与短链脂肪酸浓度成反比。我们还使用相同的逻辑构建了表达细胞因子粒细胞-巨噬细胞集落刺激因子（GM-CSF）的生物疗法。丙酸酯生物疗法在没有丙酸酯的情况下表达了大量的小鼠GM-CSF。丁酸酯生物疗法仅在动力学开始时呈现预期行为，且在没有丁酸酯的情况下反应加速。总体而言，这些遗传系统可作为测量短链脂肪酸的补充诊断工具以及生物治疗分子的递送载体。短链脂肪酸是肠道微生物群产生的关键分子。它们的浓度在某些疾病中会发生改变。在这里，我们使用逻辑“非”门和细菌启动子创建了量化丙酸酯和丁酸酯缺失的分子生物传感器。最后，我们表明这些遗传系统在缺乏这些酸的情况下可能有助于在肠道中递送治疗分子。
