Dual-RNA-seq of Acinetobacter baumannii ABC141 inside endothelial cells EA.hy926
Acinetobacter baumannii poses a substantial global health threat, causing severe multi-drug-resistant infections in hospitalized patients. Circulating clinical isolates present remarkable diversity, with a proportion capable of establishing a transient intracellular niche suitable for persistence, multiplication, and spread. Yet, it remains unknown which bacterial factors mediate the formation and maintenance of this niche, especially within non-phagocytic cells, nor what host responses are elicited. This work demonstrates that the invasive A. baumannii ABC141 strain does not secrete ammonia in endothelial cells as previously shown for other A. baumannii strains multiplying within macrophages but resides in an acidic vacuole devoid of active lysosomal degradative enzymes. Using a Dual-RNAseq approach, we mapped the host and bacterial gene expression during the replicative stage of the infection. An atypical hypoxia cell response was observed without significant induction of the HIF1 pathway, with no metabolic shift or disturbance of mitochondria. Surprisingly, ABC141 efficiently grew in hypoxic conditions in culture and within host cells. In addition, we found a bacterial signature reflective of an adaptation to a nutrient-deprived environment. Our work also highlights a differential role for ABC141 secretion systems, with the T1SS assisting intracellular multiplication and the T2SS required for host cell invasion, implicating for the first time the T2SS in the intracellular lifecycle of invasive ABC141 in endothelial cells. Overall design: Dual-RNA-seq was performed in human cells EA.hy926 infected with Acinetobacter baumannii ABC141 expressing GFP (cells were infected for 24h) to measure bacterial and human gene expression. Human cells were sorted by FACS for infected cells (using GFP to gate infected cells, Sinf) or left unsorted (Inf). Uninfected cells (sorted (SNeg) and un-sorted (Neg)) served as controls. In addition, A. baumannii ABC141 transcriptome was sequenced from the inoculum before infection of human cells (ABC141).