Nanostructured in vitro TB latency model

纳米结构体外结核潜伏期模型

• 批准号: 7737798
• 负责人: GRAHAM S TIMMINS
• 金额: $ 18.75万
• 依托单位: UNIVERSITY OF NEW MEXICO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7737798
• 关键词: Aerobic; Animal Model; Anti-Infective Agents; Area; BCG Live; Bacillus (bacterium); Bacteria; Bacterial Model; Cells; Characteristics; Chemistry; Communities; Complement; Complex; Data; Disease; Drug Delivery Systems; Drug resistance; Figs - dietary; Film; Genus Mycobacterium; Goals; Growth; Human; Hybrids; In Vitro; Infection; Knowledge; Life; Lipids; Mediating; Metabolism; Methods; Modeling; Mus; Mycobacterium tuberculosis; Nanostructures; Nanotechnology; Oxygen; Pharmaceutical Preparations; Phenotype; Physiological; Preclinical Drug Evaluation; Publications; Recovery; Research; Resistance; Science; Silicon Dioxide; Solutions; Source; Structure; Testing; Time; Tuberculosis; Tuberculosis Vaccines; Work; X ray spectroscopy; X-Ray Diffraction; drug development; frontier; high throughput screening; implantation; in vitro Model; in vivo; in vivo Model; multidisciplinary; mycobacterial; nanobiology; nanostructured; novel; prevent; public health relevance; tool

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.

描述（由申请人提供）：结核分枝杆菌潜伏在 20 亿人体内，由于重新激活而成为结核病 (TB) 的主要储存库，但潜伏杆菌的状态仍然是个谜。为了根除结核病，我们需要了解潜伏期并开发用于药物筛选的体外模型。在这个跨学科项目中，我们将微生物结核病研究与纳米技术和新型材料相结合。最近的出版物展示了混合脂质-无机纳米结构的细胞定向组装的独特现象，该现象驱动独特的非复制持久性，并保护细菌并大大延长生存能力。我们证明可以使用分枝杆菌形成这些相同的结构，并且它们表现出高度延长的潜伏期生存能力特征。我们假设潜伏结核病的持续存在及其对化疗挑战的抵抗力可以通过将杆菌纳入自组装的脂质-无机纳米结构中来模拟。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潜力。我们将建立一个新的体外模型，用于高通量筛选消除潜伏结核病的靶点和药物，以补充现有的复杂体内模型，并推动这一领域向前发展。具体目标 1 定义分枝杆菌最佳脂质-无机纳米结构包裹的条件及其对分枝杆菌耐药性的影响。具体目标 2 确定由于非复制持久性而导致的分枝杆菌代谢在脂质无机纳米结构包裹时发生的变化，并驱动体外存活和潜伏期。具体目标 3 通过显示脂质-无机纳米结构包裹的分枝杆菌在小鼠中的感染性，即使在组装后经过较长一段时间后，证明它们代表了有效的潜伏模型。 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，因为它存在于全世界 20 亿人中，并且在很大比例的病例中重新激活为活跃形式。清除这个“水库”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，但没有很好的体外潜伏期模型，但我们需要有这样一个体外模型，以便我们了解如何攻击它，也可以用来筛选药物。我们开发了一种独特的纳米结构模型，可以模拟极端的非复制持久性延迟，并希望测试它作为抗击结核病的工具的有效性。