合成生物学 ›› 2023, Vol. 4 ›› Issue (2): 263-282.DOI: 10.12211/2096-8280.2022-067
高纤云, 牛灵雪, 见妮, 管宁子
收稿日期:
2022-11-24
修回日期:
2023-01-21
出版日期:
2023-04-30
发布日期:
2023-04-27
通讯作者:
管宁子
作者简介:
基金资助:
Xianyun GAO, Lingxue NIU, Ni JIAN, Ningzi GUAN
Received:
2022-11-24
Revised:
2023-01-21
Online:
2023-04-30
Published:
2023-04-27
Contact:
Ningzi GUAN
摘要:
在人体的身体内部居住着数以万亿计的细菌和其他微生物,寄生在我们的皮肤、胃肠道和口腔等部位,微生物群失调已成为炎症性肠病、过敏、肥胖、心血管以及神经退行性疾病和癌症等多种疾病发生发展的重要原因。与此同时,利用微生物制剂改善健康状况、治疗疾病也在研究和临床中得到验证。近年来,活体生物药不断被开发用于预防、诊断与治疗疾病。采用合成生物学手段在微生物中设计、构建精准可控的基因电路,获得非致病性、具有疾病微环境感知和响应能力并按需释放治疗药物的活体工程菌,实施活体微生物疗法,已成为临床疾病治疗的新思路和新方法。本文对微生物合成生物学在疾病诊疗中的应用进展,如预防与诊疗病原菌感染、治疗代谢疾病、靶向杀伤肿瘤、缓解炎症等进行了系统性总结,并针对微生物传感器设计和推广到临床应用中面临的问题和未来的发展进行了分析与展望。利用合成生物学在微生物中设计的生物传感器能智能感知疾病微环境并可控地提供药物,在疾病的监测和治疗中有巨大的应用前景。
中图分类号:
高纤云, 牛灵雪, 见妮, 管宁子. 微生物合成生物学在疾病诊疗上的应用进展[J]. 合成生物学, 2023, 4(2): 263-282.
Xianyun GAO, Lingxue NIU, Ni JIAN, Ningzi GUAN. Applications of microbial synthetic biology in the diagnosis and treatment of diseases[J]. Synthetic Biology Journal, 2023, 4(2): 263-282.
图2 工程微生物预防与治疗病原菌感染(a)基于铜绿假单胞菌的Ⅰ型群体感应机制的基因环路。改造的大肠杆菌能合成转录因子LasR,与铜绿假单胞菌分泌的群体感应自诱导分子酰基高丝氨酸内酯(AHLs)结合,激活合成绿脓菌素裂解蛋白。(b)罗伊氏乳杆菌中基于Agr群体感应体系(agrQS)的金黄色葡萄球菌检测系统。AIP-I影响下,罗伊氏乳杆菌表达葡萄糖醛酸酶GusA被抑制,从而抑制4-硝基苯β-D-葡萄糖苷酸水解为对硝基酚(一种可以在405 nm处测量的黄色色素),通过检测吸光度变化可检测纳摩尔至微摩尔级别浓度的AIP-I。(c)基于霍乱弧菌的群体感应设计的大肠杆菌闭环识别杀伤装置。大肠杆菌分泌自诱导因子CAI-1,CAI-1激活表达YebF-Art-085使菌体裂解,释放胞内的自溶素Art-085,杀死霍乱弧菌。(d)乳酸乳球菌杂交受体CqsS-NisK的设计。杂交受体可以感应诱导因子CAI-1并触发一种易于在粪便样本中检测的酶报告基因的表达,以此检测霍乱弧菌
Fig. 2 Prevention and treatment of pathogen infection with engineered microorganisms(a) Gene circuit developed with TypeⅠquorum sensing mechanism in P. aeruginosa. The transcription factor LasR synthesized by engineered Escherichia coli can bind to acyl hyperserine lactone (AHL), a quorum sensing autoinducer secreted by P. aeruginosa, to activate the synthesis of the lysin: pseudomonectin.(b) Lactobacillus reuteri engineered with Staphylococcus aureus detection system through the Agr quorum sensing mechanism (agrQS). The expression of glucuronidase GusA is repressed by AIP-I to inhibit the enzymatic hydrolysis of 4-nitrobenzen-β-D-glucosidic acid to produce p-nitrophenol, a yellow pigment that can be detected at 405 nm. Thus, AIP-I from nanomolar to micromolar level can be detected by the change of light absorbance.(c) A closed-loop detection and killing device for Escherichia coli based on Vibrio cholerae quorum sensing. Autoinductor CAI-1 secreted by E. coli can activate the expression of YebF-Art-085 to lyse the bacteria, releasing intracellular autolysin Art-085 to kill V. cholerae.(d) Design of Lactococcus lactate hybrid receptor CqsS-NisK, which detects Vibrio cholerae by sensing inducer CAI-1 to trigger the expression of gene encode an enzyme reporter that can be easily detected in fecal samples.
图5 口服工程酿酒酵母缓解肠炎在酵母中表达高敏感型人类嘌呤能受体P2Y2,在P2Y2受体激活时分泌eATP降解酶,构建了能够感知促炎分子并依据促炎分子浓度高低智能调节的工程酵母益生菌
Fig. 5 Alleviation of enteritis by oral administration of engineered Saccharomyces cerevisiae.Highly sensitive human purinergic receptor P2Y2 is expressed in S. cerevisiae, and EATP-degrading enzyme is designed to be secreted when P2Y2 receptor is activated. Engineering S. cerevisiae is constructed to sense and regulate the pro-inflammatory molecules
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