An integrated approach using dual 13C-profiling and RNA-sequencing for unravelling the metabolic programs of the intracellular forms of Chlamydia trachomatis and their host cells

使用双 13C 分析和 RNA 测序的综合方法来揭示沙眼衣原体及其宿主细胞的细胞内代谢程序

• 批准号: 451958427
• 负责人: Professor Dr. Wolfgang Eisenreich
• 金额: --
• 依托单位: Lehrstuhl für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/451958427?language=en
• 关键词: integrated; approach; using; dual; 13C

With this proposal, we want to establish for the first time a correlation between dual RNA-seq data and isotope labelling data. Thereby, the metabolism of the human-pathogenic bacterium Chlamydia trachomatis and its host cells during the bacterial life cycle and stages shall be elucidated in unprecedented details. The study has potential to pave the way for future robust analyses of low cell numbers of C. trachomatis and subpopulations of their host cells in more advanced infection models, including suitable organoid and animal models. More specifically, the overall aim of this application is to analyse the metabolic alterations of C. trachomatis in the different intracellular states, i.e. the proliferating RBs and the non-replicating but infectious EBs as well as the persistent ARBs. This analysis will be first performed under in vitro conditions using established cell lines (e.g. HeLa) and suitable primary cell models (naïve as well as M1- and M2-polarized murine (bone-marrow) derived macrophages (BMDM), human primary macrophages, and/or fallopian tube cells). Moreover, the metabolic reprogramming especially of primary host cells infected by C. trachomatis shall be studied on a quantitative basis using the combination of dual-RNAseq and dual-isotope profiling.

通过这一提议，我们希望首次建立双RNA-seq数据和同位素标记数据之间的相关性。因此，人类致病菌沙眼衣原体及其宿主细胞在细菌生命周期和阶段的代谢将得到前所未有的详细阐明。该研究有可能为未来在更高级的感染模型（包括合适的类器官和动物模型）中对沙眼衣原体低细胞数及其宿主细胞亚群进行稳健分析铺平道路。更具体地说，本应用程序的总体目的是分析沙眼衣原体在不同细胞内状态下的代谢变化，即增殖的RBs和非复制但感染性的EBs以及持久的arb。该分析将首先在体外条件下进行，使用已建立的细胞系（例如HeLa）和合适的原代细胞模型（naïve以及M1和m2极化小鼠（骨髓）来源的巨噬细胞（BMDM），人原代巨噬细胞和/或输卵管细胞）。此外，利用双rna - naseq和双同位素分析相结合的方法，对沙眼衣原体感染的寄主细胞的代谢重编程进行定量研究。