A genome-wide in vivo screen reveals fitness pathways required for streptococcal infective endocarditis

Infective endocarditis (IE) is a life-threatening disease most often caused by blood-borne bacteria that infect previously damaged cardiac tissue. Despite the importance of this disease, the genetic basis for IE virulence remains poorly defined. Here, we present the first genome-wide in vivo analysis of bacterial fitness in a vertebrate model of IE. We identified 146 genes in Streptococcus sanguinis required for IE fitness, the majority of which had not previously been linked to endocarditis. These determinants cluster into conserved metabolic, cell envelope, transport, and regulatory pathways, representing a vast reservoir of potential targets for novel antimicrobial intervention. A subset of these genes was examined in Streptococcus mutans ; all were found to be essential for IE fitness in this distantly related oral species as well, suggesting broad conservation. Using experimental evolution, we further show that disruption of key fitness pathways triggers reproducible compensatory “bypass” mechanisms that reveal the inherent physiological constraints of the IE fitness landscape and identify vulnerable nodes for multi-target drug strategies. Together, these findings redefine streptococcal infective endocarditis as a disease shaped by conserved bacterial fitness networks that may be exploited for therapeutic development. Highlights A genome-wide in vivo screen identified 146 Streptococcus sanguinis genes required for infective endocarditis fitness. 94% of these genes represent previously unrecognized determinants of endocarditis. Multiple pathways—including CoA biosynthesis, the shikimate pathway, and rhamnan synthesis—are required for cardiac colonization. A subset of infective endocarditis fitness factors are conserved between S. sanguinis and S. mutans , with species-specific adaptations. Experimental evolution revealed compensatory metabolic networks that buffer IE fitness defects.