Metatranscriptome sequencing identifies Escherichia are major contributors to pathogenic functions and biofilm formation in diabetes related foot osteomyelitis

• Published in: Frontiers in microbiology Vol. 13; p. 956332
• Main Authors: Radzieta, Michael, Malone, Matthew, Ahmad, Mehtab, Dickson, Hugh G., Schwarzer, Saskia, Jensen, Slade O., Lavery, Lawrence A.
• Format: Journal Article
• Language: English
• Published: Frontiers Media S.A 01-08-2022
• Subjects: bone; diabetes; diabetic foot osteomyelitis (DFO); Escherichia; metatranscriptome; Microbiology; RNA-sequencing
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2022.956332

Osteomyelitis in the feet of persons with diabetes is clinically challenging and is associated with high rates of amputation. In this study RNA-sequencing was employed to explore microbial metatranscriptomes with a view to understand the relative activity and functions of the pathogen/s responsible for diabetes foot osteomyelitis (DFO). We obtained 25 intraoperative bone specimens from persons with confirmed DFO, observing that
Escherichia
spp. (7%),
Streptomyces
spp. (7%),
Staphylococcus
spp. (6%),
Klebsiella
spp. (5%) and
Proteus
spp. (5%) are the most active taxa on average. Data was then subset to examine functions associated with pathogenesis (virulence and toxins), biofilm formation and antimicrobial/multi-drug resistance. Analysis revealed
Escherichia
spp. are the most active taxa relative to pathogenic functions with K06218 (mRNA interferase
relE
), K03699 (membrane damaging toxin
tlyC
) and K03980 (putative peptidoglycan lipid II flippase
murJ
), K01114 (membrane damaging toxin plc) and K19168 (toxin cptA) being the most prevalent pathogenic associated transcripts. The most abundant transcripts associated with biofilm pathways included components of the biofilm EPS matrix including glycogen synthesis, cellulose synthesis, colonic acid synthesis and flagella synthesis. We further observed enrichment of a key enzyme involved in the biosynthesis of L-rhamnose (K01710 -dTDP-glucose 4,6-dehydratase
rfbB, rmlB, rffG
) which was present in all but four patients with DFO.