Development of Nanoparticle Based Therapies Against Tuberculosis in the Zebrafish Model, 2015

The data presented are collected through the project “Development of Nanoparticle Based Therapies Against Tuberculosis in the Zebrafish Model, 2015”. Mycobacterium tuberculosis (M.tb), the cause of tuberculosis (TB) infects one-third of the world population, one person per second, and kills about 2 million people per year. The only effective therapy, using antibiotics, is being challenged by increasing antibiotic resistance. It has been shown that when antibiotics are encapsulated in a biodegradable nanoparticle (NP) made from a polymer such as poly (lactate-co-glycolate) acid (PLGA) the drug is significantly more effective in curing TB in different animal models; higher concentrations of antibiotics can be targeted selectively to macrophages, the infection site, where they are released slowly, while the total systemic levels is much less than with standard drug therapy. A weakness of the existing animal models of TB is the difficulty in visualizing the NPs in the infected live animal, which is essential if one is to optimize beads for their ability to reach infected cells after different modes of administration. This is precisely the great strength of the transparent zebrafish model of tuberculosis based on red and green variants of M.marinum, the fish TB organism. In this project a model using red and green fluorescent M. marinum was established, and it also successfully fabricated PLGA NPs enclosing the antibiotic rifampicin and red or green fluorescent dyes. When these beads are injected into embryos 1-2 days after infection they reach the same cells in the granulomas as the bacteria and we have preliminary data giving hints of therapy. Furthermore the aim is now to perfect this system for embryos and adults and find the most effective NPs that reach granulomas and provide a robust therapy. This system enables to screen for different kinds of NPs, as well as to compare different antibiotics and combinations thereof, and cDNA's coding for anti-microbial peptides. The most effective NPs will be given to collaborators in India to test them in a guinea pig model as a prelude to testing the NPs in humans.

本数据集收录的数据源自2015年“基于纳米颗粒的斑马鱼模型抗结核治疗开发”项目。结核分枝杆菌（Mycobacterium tuberculosis, M.tb）是结核病（tuberculosis, TB）的致病菌，全球约三分之一人口曾受其感染，每秒钟即有1人被感染，每年约造成200万人死亡。目前唯一有效的治疗手段为抗生素疗法，但日益严峻的抗生素耐药性问题正对其构成挑战。已有研究证实，将抗生素封装于聚乳酸-羟基乙酸共聚物（poly(lactate-co-glycolate) acid, PLGA）等聚合物制备的可生物降解纳米颗粒（nanoparticle, NP）中时，该药物在多种动物模型的结核治疗中效果显著提升：抗生素可选择性靶向感染部位的巨噬细胞，缓慢释放药物并维持较高局部浓度，而全身药物暴露水平远低于标准药物疗法。
现有结核动物模型的一大缺陷是难以在感染活体动物中可视化追踪纳米颗粒，而若要优化纳米颗粒经不同给药方式后抵达感染细胞的能力，该可视化追踪环节必不可少。而基于海分枝杆菌（M.marinum，鱼类结核致病菌）红绿荧光变体的透明斑马鱼结核模型，恰好解决了这一难题。本项目已成功构建携带红绿荧光标记的海分枝杆菌感染模型，同时制备出包裹抗生素利福平（rifampicin）以及红绿荧光染料的PLGA纳米颗粒。将此类纳米颗粒注射至感染1~2天的斑马鱼胚胎后，它们可与细菌一同抵达肉芽肿内的相同细胞区域，我们已获得初步数据，为该治疗策略提供了佐证。
本项目的后续目标为优化该系统，使其适用于斑马鱼胚胎与成体，并筛选出可有效抵达肉芽肿、实现可靠治疗效果的最优纳米颗粒。该系统可用于筛选不同类型的纳米颗粒，同时对比不同抗生素及其联合疗法，以及编码抗菌肽的互补脱氧核糖核酸（complementary DNA, cDNA）的治疗效果。研究中筛选出的最优纳米颗粒将交由印度的合作方，在豚鼠（guinea pig）模型中开展测试，以此作为人体试验前的预实验。