PO.PR01.04 · 预防研究
利用表达导向抗菌肽（gAMPs）的生物工程益生菌针对 CRC 相关具核梭杆菌的精准治疗
Precision therapy against CRC associated Fusobacterium nucleatum using bioengineered probiotics expressing guided antimicrobial peptides (gAMPs)
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摘要 Abstract
中文摘要
结直肠癌（CRC）是癌症相关死亡的主要原因之一，具核梭杆菌（Fusobacterium nucleatum，F. nucleatum）被确定为其进展的关键促成因素。本研究探索了一种新型疗法，通过使用一种表达导向抗菌肽（gAMPs）的生物工程益生菌来选择性抑制 F. nucleatum，从而靶向这一病原体。在本研究中，将 Lactococcus lactis MG1363 进行工程化改造，使其表达源自 Ovispirin 和 Cathelin 相关肽 SCF 的 gAMPs，并连接到一个源自 Statherin 的导向肽上，该导向肽可特异性结合 F. nucleatum 的膜孔蛋白 FomA。该细菌在 nisin 诱导 PNisA 启动子的作用下表达 AMP/gAMP，并通过胞外分泌信号 usp45 将其分泌。对所得的合成肽和益生菌进行了针对靶标 F. nucleatum 和其他非靶标细菌的抗菌活性测定。用合成肽在体外或用益生菌与多微生物群落共培养进行了生物膜抑制和生长动力学测定。我们发现，源自 Statherin 的导向肽增强了对 F. nucleatum 的结合亲和力，与对照肽相比显著增加了附着。体外测定揭示，无导向和有导向的 AMP 均有效抑制了 F. nucleatum 的生物膜形成，其中 gAMPs 对非靶标细菌的毒性降低。gAMPs 在调节生长动力学方面更为有效，在较低浓度下表现出对 F. nucleatum 的选择性毒性。在模拟人类肠道微生物组中的共培养实验显示，gAMP 益生菌在有效降低 F. nucleatum 丰度的同时维持了微生物多样性。定量 PCR 和 16S rRNA 测序证实，gAMP 治疗保留了微生物群的丰富度，这与对照样本中观察到的显著菌群失调形成对比。这些发现支持工程化益生菌作为靶向 CRC 相关 F. nucleatum 的治疗方法的潜力。
查看英文原文 English abstract
Colorectal cancer (CRC) is a leading cause of cancer-related mortality, with Fusobacterium nucleatum (F. nucleatum ) identified as a key contributor to its progression. This study explores a novel therapy that targets this pathogen by using a bioengineered probiotic that expresses guided antimicrobial peptides (gAMPs) to selectively inhibit F. nucleatum. in this study, Lactococcus lactis MG1363 was engineered to express gAMPs derived from Ovispirin and Cathelin-related peptide SCF, linked to a Statherin-derived guide peptide that binds specifically to the F. nucleatum membrane porin FomA. The bacteria expressed the AMP/gAMP under the induction of the PNisA promoter by nisin and secreted it via the extracellular secretion signal usp45. The resultant synthetic peptides and probiotics were assayed for antimicrobial activity against the targeted F. nucleatum and other non-target bacteria. Biofilm inhibition and growth kinetic assays were performed with synthetic peptides in vitro or the probiotic in co-culture with a polymicrobial community. We saw that statherin-derived guide peptide enhanced the binding affinity to F. nucleatum , significantly increasing attachment compared to control peptides. In vitro assays revealed that both unguided and guided AMPs effectively inhibited biofilm formation in F. nucleatum , with gAMPs showing reduced toxicity against non-target bacteria. The gAMPs were more effective in modulating growth kinetics, exhibiting selective toxicity towards F. nucleatum at lower concentrations. Co-culture experiments in a simulated human gut microbiome showed the gAMP probiotic maintained microbial diversity while effectively reducing F. nucleatum abundance. Quantitative PCR and 16S rRNA sequencing confirmed that gAMP treatment preserved the richness of the microbiota, contrasting with significant dysbiosis observed in control samples. These findings support the potential of engineered probiotics as a therapeutic approach that targets CRC-associated F. nucleatum .
利益披露 Disclosure
A. Choudhury, None..
C. Scano, None..
A. Barton, None.