Nanostructured in vitro TB latency model

描述(申请人提供)：结核分枝杆菌潜伏在20亿人中，由于重新激活而成为结核病(TB)的主要宿主，但潜伏杆菌的状态仍然是一个谜。为了根除结核病，我们需要了解潜伏期并开发药物筛选的体外模型。在这个跨学科的项目中，我们将微生物结核病研究与纳米技术和新材料相结合。最近发表的文章显示了一种独特的现象，即细胞定向组装的杂化脂质-无机纳米结构，驱动独特的非复制持久性，并保护细菌和显着延长生存能力。我们表明，这些相同的结构可以使用分枝杆菌形成，并且它们显示出潜伏期的高度延长的生存特性。我们假设，潜伏结核的持久性及其对化疗挑战的抵抗力，可以通过将杆菌掺入自组装的脂类-无机纳米结构来模拟。该模型结合了体外研究和生长条件调节的能力，以及体内植入和感染的可能性。我们将建立一个新的体外模型，用于高通量筛选靶点和可以消除潜伏性结核病的药物，以补充现有的复杂的体内模型，并允许该领域向前发展。具体目标1确定分枝杆菌最佳脂质-无机纳米结构包埋的条件及其对分枝杆菌耐药性的影响。特定目的2确定包裹在非复制持久性导致的脂类-无机纳米结构中的分枝杆菌代谢的变化，并驱动体外存活和潜伏期。具体目标3表明，脂类-无机纳米结构包裹的分枝杆菌通过显示它们在小鼠中的传染性，代表了潜伏期的有效模型，即使在组装后的较长时间之后也是如此。 公共卫生相关性：公共卫生相关性我们需要了解结核病能够以潜伏状态存在的方式，因为它存在于全球20亿人中，并在相当大比例的病例中重新激活为活跃形式。清除这一“蓄水池”将是最终根除战略的重要组成部分。虽然有潜伏期的动物模型，但没有良好的体外潜伏期模型，但我们需要有这样的体外模型，以便我们能够了解如何攻击它，也可以用来筛选药物。我们已经开发了一种独特的纳米结构模型，它模拟了潜伏期的极端非复制性持久性，并希望测试它作为抗击结核病的工具可能有多有效。

DESCRIPTION (provided by applicant): Mycobacterium tuberculosis latently persist in 2 billion people, representing a major reservoir of tuberculosis (TB) due to reactivation, yet the state of latent bacilli remains a mystery. In order to eradicate TB, we need to understand latency and develop in vitro models for drug screening. In this interdisciplinary project we have merged microbiological tuberculosis research with nanotechnology and novel materials. Recent publications demonstrate a unique phenomenon of cell-directed assembly of hybrid lipid-inorganic nanostructures that drive a unique non-replicative persistence, and protect bacteria and drastically prolong viability. We show these same structures can be formed using mycobacteria, and that they display the highly extended viability characteristic of latency. We hypothesize that the persistence of latent TB, and its resistance to chemotherapeutic challenge, can be modeled by incorporation of bacilli into self-assembled lipid-inorganic nanostructures. This model combines the ability for in vitro study and growth condition modulation, with the potential for in vivo implantation and infection. We will generate a new in vitro model for high throughput screening for targets of, and drugs that can eliminate latent TB, to complement the existing complex in vivo models and al- low this field to move forward. Specific Aim 1 Define the conditions for optimal lipid-inorganic nanostructure-encasement of mycobacteria and its effects upon mycobacterial resistance to drugs. Specific Aim 2 Determine the changes in mycobacterial metabolism occurring upon encasement in lipid- inorganic nanostructures that result from non-replicative persistence, and drive in vitro survival and latency. Specific Aim 3 Demonstrate that lipid-inorganic nanostructure-encased mycobacteria represent valid models of latency by showing their infectivity in mice, even after extended periods of time after assembly. PUBLIC HEALTH RELEVANCE: Public Health Relevance We need to understand the way that tuberculosis is able to exist in a latent state because it is present in 2 billion people worldwide and reactivates into an active form in a significant percentage of cases. Clearing this 'reservoir' will be an essential component of an eventual eradication strategy. Although there are animal models of latency, there is no good model of in vitro latency, yet we need to have such an in vitro model so that we can understand how to attack it, and also to screen drugs with. We have developed a unique nanostructured model that mimics the extreme non-replicative persistence of latency, and want to test how effective it might be as a tool in the battle against TB.