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

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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.