A Bioengineered Three-Dimensional Cell Culture Platform Integrated with Microfluidics To Address Antimicrobial Resistance in Tuberculosis.
A Bioengineered Three-Dimensional Cell Culture Platform Integrated with Microfluidics To Address Antimicrobial Resistance in Tuberculosis.
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DOI：
10.1128/mbio.02073-16
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发表时间：
2017-02-07
期刊：
影响因子：
6.4
通讯作者：
Elkington P
中科院分区：
文献类型：
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作者：
Bielecka MK;Tezera LB;Zmijan R;Drobniewski F;Zhang X;Jayasinghe S;Elkington P
随着完全耐药生物体的出现，抗菌素耐药性是对人类健康的最重大威胁之一。我们结合了生物工程、转基因细菌、纵向读数和流体学，开发了一个变革性平台，以解决药物开发瓶颈，利用结核分枝杆菌作为模式生物。我们通过生物电喷雾方法产生了将毒性报告杆菌、原代人细胞和细胞外基质结合的微球。肉芽肿在三维基质内形成，并且分枝杆菌应激基因上调。吡嗪酰胺是治疗人类结核病的重要一线抗生素，可杀死M。在三维培养物中而不是在标准的二维培养物或Middlebrook 7H9肉汤中检测到结核病，表明微球内的抗生素敏感性反映了患者的状况。然后，我们通过将微球系统与微流控板相结合来进行药代动力学建模，并证明我们可以模拟动态抗生素浓度对分枝杆菌杀灭的影响。微球系统是高度易处理的，允许细胞含量、细胞外基质、球体大小、感染剂量和周围介质的变化，具有解决广泛的人类感染和抗微生物耐药性威胁的潜力。抗生素耐药性是一个主要的全球性威胁，一个新兴的概念是，感染应在宿主免疫细胞的背景下进行研究。结核病是一种慢性感染，每年造成100多万人死亡，并且对抗生素的耐药性越来越强。最近关于缩短治疗时间或新疫苗接种方法的主要研究并不成功，这表明需要变革性技术来控制结核病。我们已经开发了一个全新的系统，通过使用生物工程来研究三维基质中宿主细胞的感染。我们发现，对患者有效的抗生素在这种微球系统中有效，但在标准感染系统中无效。然后，我们将微球与微流体相结合，以模拟患者体内药物浓度的变化，并证明增加抗生素浓度对细菌存活的影响。该系统可广泛应用于解决抗生素耐药性的威胁和开发新的治疗方法。
Antimicrobial resistance presents one of the most significant threats to human health, with the emergence of totally drug-resistant organisms. We have combined bioengineering, genetically modified bacteria, longitudinal readouts, and fluidics to develop a transformative platform to address the drug development bottleneck, utilizing Mycobacterium tuberculosis as the model organism. We generated microspheres incorporating virulent reporter bacilli, primary human cells, and an extracellular matrix by using bioelectrospray methodology. Granulomas form within the three-dimensional matrix, and mycobacterial stress genes are upregulated. Pyrazinamide, a vital first-line antibiotic for treating human tuberculosis, kills M. tuberculosis in a three-dimensional culture but not in a standard two-dimensional culture or Middlebrook 7H9 broth, demonstrating that antibiotic sensitivity within microspheres reflects conditions in patients. We then performed pharmacokinetic modeling by combining the microsphere system with a microfluidic plate and demonstrated that we can model the effect of dynamic antibiotic concentrations on mycobacterial killing. The microsphere system is highly tractable, permitting variation of cell content, the extracellular matrix, sphere size, the infectious dose, and the surrounding medium with the potential to address a wide array of human infections and the threat of antimicrobial resistance. Antimicrobial resistance is a major global threat, and an emerging concept is that infection should be studied in the context of host immune cells. Tuberculosis is a chronic infection that kills over a million people every year and is becoming progressively more resistant to antibiotics. Recent major studies of shorter treatment or new vaccination approaches have not been successful, demonstrating that transformative technologies are required to control tuberculosis. We have developed an entirely new system to study the infection of host cells in a three-dimensional matrix by using bioengineering. We showed that antibiotics that work in patients are effective in this microsphere system but not in standard infection systems. We then combined microspheres with microfluidics to model drug concentration changes in patients and demonstrate the effect of increasing antibiotic concentrations on bacterial survival. This system can be widely applied to address the threat of antimicrobial resistance and develop new treatments.