Metagenome Data Mining to Elucidate Prevalence and Virulence Potentials of Biosafety-level-2 Microorganisms from ISS Environments

In late 2018, our NASA collaborators, Venkateswaran and Singh, reported the proliferation of the dominant BSL-2 pathogen, Klebsiella pneumoniae, on ISS surfaces over a three flight time course (1). Here we propose to utilize the same GeneLab dataset (GLDS-69), supplemented with a ground-based mutigenerational cultivation study of ISS K. pneumoniae isolates, to gain a better understanding of the threat these opportunistic pathogens may pose to astronaut health. This overarching objective will be achieved by mining GLDS-69 to reconstruct metagenome-assembled genomes (MAGs) for K. pneumoniae – using these to assess strain-level diversity and potential changes in pathogenic potential. In parallel, K. pneumoniae isolates from the JPL ISS culture collection will be subjected to multigenerational selection using stressors consistent with extended space travel and tracked for changes in gene expression patterns (e.g. virulence gene complement). Selection in the laboratory (e.g. directed evolution) will be conducted in the presence of disinfectants used on the ISS, followed by a survey of gene expression of evolved and unevolved strains under simulated microgravity. The proposed project will be conducted in collaboration with JPL and GeneLab scientists, to gain a better understanding of strain-level succession of a dominant potential pathogen on the ISS and assess potential stimulation of virulence factors under selective pressure from relevant disinfectants and microgravity.

This is a mature technology (TRL 7+) — the realistic path in is usually NASA's Technology Transfer Program: licensing an existing NASA patent, or a Space Act Agreement to use NASA facilities/expertise directly. NASA also runs a startup licensing program with no upfront fee for companies formed to commercialize a specific NASA technology.

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