# Transcriptomic signatures of host immune responses in aphthous ulcers, the earliest lesions of Crohn's disease, suggest that bacterial uptake, rather than global dysbiosis, is the initiating factor

## TL;DR

The study suggests that Crohn's disease starts with bacterial uptake triggering immune responses, not from a general gut bacteria imbalance.

## Contribution

The study identifies bacterial uptake and immune activation as the initiating factor in Crohn's disease, rather than global gut dysbiosis.

## Key findings

- Aphthous ulcers show gene signatures of epithelial stress, antimicrobial defense, and plasma cell activation.
- Monocyte recruitment and interferon-γ signaling are prominent in early Crohn's lesions.
- Normal mucosa adjacent to ulcers resembles healthy tissue, indicating localized immune activation.

## Abstract

Crohn's disease is a chronic, transmural inflammatory disease of the human gut. Changes in the fecal microbial composition and dysbiosis are consistent features in studies of Crohn's disease patients, but whether dysbiosis is a cause or consequence of inflammation remains unresolved. Genetic susceptibility plays a role in the development of Crohn's disease and has been linked to genes involved in recognition of intestinal bacteria by the mononuclear phagocyte system. The earliest visible lesions in Crohn's disease are aphthous ulcers, overlying Peyer's patches and lymphoid follicles. To identify mechanisms underlying the earliest stages of disease we compared gene expression in aphthous ulcers, Peyer's patches, inflamed and endoscopically normal mucosa from patients and controls using total RNA‐seq. The resulting data were subjected to network analysis to identify coregulated gene expression signatures of cell types and processes. These results were compared to single‐cell RNA‐seq analysis of intestinal macrophages in normal and diseased mucosa. The analysis of aphthous ulcers revealed signatures of epithelial stress and antimicrobial defense, plasma cell activation and immunoglobulin production, monocyte recruitment, inflammatory gene expression and induction of interferon‐γ. These signatures were not present in the normal appearing mucosa adjacent to aphthous ulcers, which were similar to healthy control mucosa. Given the role of Peyer's patches and lymphoid follicles in sampling the luminal contents, these findings suggest the initial lesion in Crohn's disease arises from the uptake of bacteria and the activation of multiple host defense pathways rather than the breakdown of epithelial barrier integrity and widespread bacterial translocation.

To identify mechanisms underlying the earliest stages of disease initiation, we compared gene expression in aphthous ulcers, Peyer's patches, inflamed and endoscopically normal mucosa from patients and controls using total RNA‐seq. The signatures obtained suggest epithelial stress, plasma cell activation, monocyte activation and interferon‐γ signaling are driving the dysregulated response. We conclude that the initial lesion in Crohn's disease arises from an invasive bacterial challenge leading to intense activation of multiple host defense pathways rather than the breakdown of epithelial barrier integrity and widespread bacterial translocation.