The Burkholderia pseudomallei intracellular 'TRANSITome'

Area of Science:

• Microbiology
• Genomics
• Infectious Diseases

Background:

• Prokaryotic cell transcriptomics traditionally analyzes mixed populations, limiting understanding of individual cell behavior.
• Spatiotemporal and stage-specific processes in complex environments remain poorly understood.
• Single-cell analysis is crucial for dissecting heterogeneous prokaryotic populations.

Purpose of the Study:

• To develop a TRANSITomic approach for single-cell transcriptomics in prokaryotes.
• To profile the transcriptome of Burkholderia pseudomallei during host cell infection.
• To gain pathophysiological insights into host-pathogen interactions at a single-cell level.

Main Methods:

• Development of the 'TRANSITomic' approach for single-cell transcriptomics.
• Profiling transcriptomes of individual Burkholderia pseudomallei cells during host cell infection.
• Identification of gene expression dynamics and virulence factors.

Main Results:

• Identified three distinct stages of Burkholderia pseudomallei transit within host cells: vacuole entry, cytoplasmic escape/replication, and membrane protrusion for cell-to-cell spread.
• Revealed dynamic gene expression flux during host cell transit.
• Identified essential genes and hypothetical proteins involved in virulence, including attachment, cytoskeletal modulation, and autophagy evasion.

Conclusions:

• The TRANSITomic approach enables high-resolution prokaryotic single-cell transcriptomics.
• Provides a detailed understanding of Burkholderia pseudomallei infection dynamics and virulence mechanisms.
• Advances the study of host-pathogen interactions by offering single-cell resolution.