Three-dimensional analysis and modeling of the Chlamydia developmental cycle

衣原体发育周期的三维分析和建模

• 批准号: 9035928
• 负责人: Ming Tan
• 金额: $ 18.44万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035928
• 关键词: Area; Bacteria; Binding; Body Size; Cell Fate Control; Cells; Centers for Disease Control and Prevention (U.S.); Chlamydia; Chlamydia Infections; Communicable Diseases; Computing Methodologies; Confocal Microscopy; Crowding; Data; Data Analyses; Development; Developmental Process; Disease Notification; Electron Microscopy; Environment; Equilibrium; Event; Future; Genital system; Hour; Human; Image; Individual; Infection; Infectious Agent; Location; Lung diseases; Membrane; Modeling; Process; Production; Property; Public Health; Regulation; Reporting; Scanning; Scanning Electron Microscopy; Sexually Transmitted Diseases; Surface; Therapeutic; Thick; Three-Dimensional Image; Three-dimensional analysis; Time; base; extracellular; genital infection; innovation; mathematical model; microscopic imaging; monolayer; novel; novel strategies; pathogen; pathogenic bacteria; public health relevance; rate of change; reconstruction; three-dimensional modeling

 DESCRIPTION (provided by applicant): Chlamydia is one of the most important infectious agents from a public health perspective. In 2011 more than 1.4 million cases of chlamydial infections were reported to the CDC making it the most commonly reported infectious disease in the U.S. Chlamydia causes an intracellular infection that is unusual among pathogenic bacteria because it involves conversion between two specialized forms of the bacterium. The reticulate body (RB) is an intracellular form that replicates by binary fission, while the elementary body (EB) is the infectious form that can transmit the infection to a new cell. A successful infectious cycle involves both RB replication and RB-to-EB conversion within an intracellular compartment called the chlamydial inclusion, but it is not known how these processes are regulated. We have developed an innovative approach to obtain detailed three dimensional views of a Chlamydia-infected cell. We first perform electron microscopy imaging on serial sections through the cell, then use computational methods to reconstruct these scans into a 3D image and finally trace the outline of each of the bacteria. With this approach we can visualize the entire chlamydial inclusion and the numbers and locations of all the RBs and EBs. In Aim 1 of this application, we propose to obtain and compare 3D reconstructions of Chlamydia-infected cells over the course of the 48-hour developmental cycle. In Aim 2, we will analyze this data with mathematical and modeling approaches to determine if the proportion of RBs that are replicating or converting changes with time. We will also examine if RB replication and RB-to-EB conversion correlate with external factors such as the size of the inclusion or the surface area of the inclusion membrane. This novel approach will provide an unprecedented level of quantitative and spatial detail about a Chlamydia-infected cell. This information will help us to understand fundamental properties about the intracellular infection, including how this important pathogen replicates and produces infectious progeny inside a human cell.

 描述（由适用提供）：从公共卫生的角度来看，衣原体是最重要的传染性药物之一。在2011年，疾病预防控制中心报告了超过140万例衣原体感染病例，这使其成为美国衣原体最常见的传染病，引起了病原细菌中不寻常的细胞内感染，因为它涉及两种细菌的​​两种专业形式之间的转化。网状体（RB）是通过二元裂变复制的细胞内形式，而基本体（EB）是可以将感染传播到新细胞的传染性形式。成功的感染周期涉及RB复制和RB-eb-eb转换，它在称为衣原体包容的细胞内室内，但尚不清楚如何调节这些过程。我们已经开发了一种创新的方法，以获取感染衣原体的细胞的详细三维观点。我们首先通过细胞对串行切片进行电子显微镜成像，然后使用计算方法将这些扫描重建为3D图像，并最终追踪每个细菌的轮廓。通过这种方法，我们可以将整个衣原体包容性以及所有RB和EB的数量和位置可视化。在本应用的AIM 1中，我们建议在48小时发育周期中获得并比较感染衣原体感染细胞的3D重建。在AIM 2中，我们将使用数学和建模方法分析这些数据，以确定正在复制或转换随时间变化的RB的比例。我们还将检查RB复制和Rb-to-eB转换是否与外部因素（例如纳入膜的大小或表面积）相关。这种新颖的方法将提供有关感染衣原体的细胞的前所未有的定量和空间细节。这些信息将帮助我们了解有关细胞内感染的基本特性，包括该重要病原体如何复制并在人类细胞内部产生传染性后代。