Dataset Information

Mass Spectrometry-Based Proteomic Profiling of Sonicate Fluid Differentiates Staphylococcus aureus Periprosthetic Joint Infection from Non-Infectious Failure: A pilot study

ABSTRACT: This study aims to use proteomic profiling of sonicate fluid samples to compare host response during Staphylococcus aureus-associated periprosthetic joint infection (PJI) and non-infected arthroplasty failure (NIAF) and investigate novel biomarkers to increase diagnostic accuracy. In this pilot study, eight sonicate fluid samples (four from NIAF and four from caused by Staphylococcus aureus PJI) were studied. Samples were reduced, alkylated and trypsinized overnight, followed by analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) on a high-resolution Orbitrap Eclipse mass spectrometer. MaxQuant software suite was used for protein identification, filtering, and label free quantitation. Principal component analysis of the identified proteins clearly separated S. aureus PJI and NIAF samples. Overall, 810 proteins were quantified in any three samples from each group and 35 statistically significant differentially abundant proteins (DAPs) were found (2-sample t-test p-values ≤0.05 and log2fold-change values ≥2 or ≤-2). Gene ontology pathway analysis found that microbial defense responses, specifically those related to neutrophil activation, were increased in S. aureus PJI compared to NIAF samples. Proteomic profiling of sonicate fluid using LC-MS/MS, alone or in combination with complementary protein analyses, differentiated S. aureus PJI and NIAF in this pilot study.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Akhilesh Pandey

LAB HEAD: Akhilesh Pandey

PROVIDER: PXD038928 | Pride | 2023-10-24

REPOSITORIES: Pride

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Publications

Mass spectrometry-based proteomic profiling of sonicate fluid differentiates Staphylococcus aureus periprosthetic joint infection from non-infectious failure: A pilot study.

Fisher Cody R CR Mangalaparthi Kiran K KK Greenwood-Quaintance Kerryl E KE Abdel Matthew P MP Pandey Akhilesh A Patel Robin R

Proteomics. Clinical applications 20230327 5

<h4>Purpose</h4>This pilot study aimed to use proteomic profiling of sonicate fluid samples to compare host response during Staphylococcus aureus-associated periprosthetic joint infection (PJI) and non-infected arthroplasty failure (NIAF) and identify potential novel biomarkers differentiating the two.<h4>Experimental design</h4>In this pilot study, eight sonicate fluid samples (four from NIAF and four from S. aureus PJI) were studied. Samples were reduced, alkylated, and trypsinized overnight, ...[more]

PMID: 36938941

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Project description:The opportunistic pathogen, Staphylococcus aureus, encounters a wide variety of fluid shear levels within the human host, which may play a key role in dictating whether this organism adopts a commensal interaction with the host or transitions to cause disease. Using rotating-wall vessel bioreactors to create a physiologically-relevant, low fluid shear environment, S. aureus was evaluated for cellular responses that could impact its colonization and virulence. S. aureus cells grown in a low fluid shear environment initiated a novel attachment-independent biofilm phenotype and were completely encased in extracellular polymeric substances. Compared to controls, low-shear cultured cells displayed slower growth and repressed virulence characteristics, including decreased carotenoid production, increased susceptibility to oxidative stress, and reduced survival in whole blood. Transcriptional whole genome microarray profiling suggested alterations in metabolic pathways. Further genetic expression analysis revealed the down-regulation of the RNA chaperone Hfq, which parallels low fluid shear responses of certain Gram negative organisms. This is the first study to report an Hfq association to fluid shear in a Gram positive organism, suggesting an evolutionarily conserved response to fluid shear among structurally diverse prokaryotes. Collectively, our results suggest S. aureus responds to a low fluid shear environment by initiating a biofilm/colonization phenotype with diminished virulence characteristics, which could lead to insight into key factors influencing the divergence between infection and colonization during initial host pathogen interaction. Genetic expression profiles of Staphylococcus aureus cultured under low fluid shear conditions was compared to control cultures of S. aureus which was cultured in identical hardware in an orientation disrupting the low fluid shear effect. Samples from the same date of culture were compared (control 21:low 21 and control 30: low 30). S. aureus was cultured for 20 hours in either the low fluid shear or control orientated rotating wall vessel (RWV) bioreactor at which point the cells were removed and RNA extracted. At 20 hours, both cultures were in the same stage of growth (stationary phase) and at this point phenotypic differences between control and low fluid shear cultures were noted.

Dataset Information

Mass Spectrometry-Based Proteomic Profiling of Sonicate Fluid Differentiates Staphylococcus aureus Periprosthetic Joint Infection from Non-Infectious Failure: A pilot study

Publications

Mass spectrometry-based proteomic profiling of sonicate fluid differentiates Staphylococcus aureus periprosthetic joint infection from non-infectious failure: A pilot study.

Similar Datasets

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