Simultaneous profiling of host expression and microbial abundance by spatial metatranscriptome sequencing

We developed an analysis pipeline that can extract microbial sequences from spatial transcriptomic (ST) data and assign taxonomic labels, generating a spatial microbial abundance matrix in addition to the default host expression matrix, enabling simultaneous analysis of host expression and microbial distribution. We called the pipeline spatial metatranscriptome (SMT) and applied it on both human and murine intestinal sections and validated the spatial microbial abundance information with alternative assays. Biological insights were gained from these novel data that showed host-microbe interaction at various spatial scales. Finally, we tested experimental modification that can increase microbial capture while preserving host spatial expression quality and, by use of positive controls, quantitatively showed the capture efficiency and recall of our methods. This proof-of-concept work shows the feasibility of SMT analysis and paves the way for further experimental optimization and application.

Automated summary

We developed an analysis pipeline called spatial metatranscriptome (SMT) that extracts microbial sequences from spatial transcriptomic (ST) data, assigns taxonomic labels, and generates a spatial microbial abundance matrix. The pipeline was applied to human and murine intestinal sections and validated with alternative assays. This work provides biological insights into host-microbe interactions at different spatial scales and demonstrates the feasibility of SMT analysis.