Fecal Host Transcriptomics for Non-Invasive Human Mucosal Immune Profiling: Proof of Concept in Clostridium Difficile...

Area of Science:

• Microbiology
• Immunology
• Genomics

Background:

• Host factors significantly influence Clostridium difficile infection (CDI) pathogenesis and outcomes.
• Characterizing host responses may reveal biomarkers to supplement current microbe-based diagnostics.

Purpose of the Study:

• To investigate host fecal transcriptomics for non-invasive profiling of mucosal immune responses in CDI.
• To identify potential host-based biomarkers for CDI diagnosis and understanding disease mechanisms.

Main Methods:

• Fecal samples from CDI patients and non-CDI diarrhea controls were collected.
• Human mRNA was profiled using amplicon-based next-generation sequencing (NGS).
• Fecal host mRNA expression profiles were compared between CDI patients and controls.

Main Results:

• The ratio of human actin gamma 1 (ACTG1) to 16S ribosomal RNA (rRNA) correlated with NGS quality.
• Principal component analysis differentiated CDI patients from controls based on fecal mRNA profiles.
• Differentially expressed genes related to immune response (e.g., IL23A, IL34) and actin-cytoskeleton function were identified.

Conclusions:

• Fecal transcriptomics offers a non-invasive method for profiling host immune responses in CDI.
• Identified differentially expressed genes show biological plausibility, supporting their role in CDI.
• This approach demonstrates the potential of fecal transcriptomics for discovering host-based biomarkers in enteric infections.