Microbial Forensics: Predicting Phenotypic Characteristics and Environmental Conditions from Large-Scale Gene Expression Profiles

A tantalizing question in cellular physiology is whether the cellular state and environmental conditions can be inferred by the expression signature of an organism. To investigate this relationship, we created an extensive normalized gene expression compendium for the bacterium Escherichia coli that was further enriched with meta-information through an iterative learning procedure. We then constructed an ensemble method to predict environmental and cellular state, including strain, growth phase, medium, oxygen level, antibiotic and carbon source presence. Results show that gene expression is an excellent predictor of environmental structure, with multi-class ensemble models achieving balanced accuracy between 70.0% (±3.5%) to 98.3% (±2.3%) for the various characteristics. Interestingly, this performance can be significantly boosted when environmental and strain characteristics are simultaneously considered, as a composite classifier that captures the inter-dependencies of three characteristics (medium, phase and strain) achieved 10.6% (±1.0%) higher performance than any individual models. Contrary to expectations, only 59% of the top informative genes were also identified as differentially expressed under the respective conditions. Functional analysis of the respective genetic signatures implicates a wide spectrum of Gene Ontology terms and KEGG pathways with condition-specific information content, including iron transport, transferases, and enterobactin synthesis. Further experimental phenotypic-to-genotypic mapping that we conducted for knock-out mutants argues for the information content of top-ranked genes. This work demonstrates the degree at which genome-scale transcriptional information can be predictive of latent, heterogeneous and seemingly disparate phenotypic and environmental characteristics, with far-reaching applications.Author Summary: The transcriptional profile of an organism contains clues about the environmental context in which it has evolved and currently lives, its behavior and cellular state. It is yet unclear, however, how much information can be efficiently extracted and how it can be used to classify new samples with respect to their environmental and genetic characteristics. Here, we have constructed an extensive transcriptome compendium of Escherichia coli that we have further enriched via an iterative learning approach. We then apply an ensemble of various machine learning algorithms to infer environmental and cellular information such as strain, growth phase, medium, oxygen level, antibiotic and carbon source. Functional analysis of the most informative genes provides mechanistic insights and palpable hypotheses regarding their role in each environmental or genetic context. Our work argues that genome-scale gene expression can be a multi-purpose marker for identifying latent, heterogeneous cellular and environmental states and that optimal classification can be achieved with a feature set of a couple hundred genes that might not necessarily have the most pronounced differential expression in the respective conditions.