Session: 868 APS Barrier Function and Repair Poster Session
(868.4) N6-Methyladenosine (m6A) Modifies Regenerative Transcripts in the Intestinal Epithelium
Tuesday, April 5, 2022
10:15 AM – 12:15 PM
Location: Exhibit/Poster Hall A-B - Pennsylvania Convention Center
Poster Board Number: E212
Charles Danan (Childrens Hospital of Philadelphia), Katharina Hayer (Childrens Hospital of Philadelphia), Lauren Simon (Childrens Hospital of Philadelphia), Matthew Weitzman (Childrens Hospital of Philadelphia), Kathryn Hamilton (Childrens Hospital of Philadelphia)
Presenting Author Childrens Hospital of Philadelphia
Background. The intestinal epithelium forms a critical barrier between foreign antigens in the intestinal lumen and immune cells in the underlying mucosa. This barrier is compromised in inflammatory bowel disease (IBD), resulting in inflammation and epithelial cell death. Improved understanding of intestinal epithelial regeneration would identify novel therapeutic targets to enhance epithelial barrier repair in diseases such as IBD. After injury, tissues such as the blood, skeletal muscle, and central nervous system activate a regenerative program that is partially regulated by N6-methyladeonsine (m6A) modification of RNA. However, few data exist regarding the role of m6A within the intestinal epithelium. The goal of this study was to profile m6A-modification transcriptome-wide in the regenerating intestinal epithelium. We hypothesize that after injury, m6A promotes regeneration within the intestinal epithelium by modifying transcripts important in epithelial repair.
Methods. Wildtype mice were given dextran sodium sulfate (DSS) in drinking water to induce intestinal inflammation and epithelial cell loss. Control mice were provided untreated water. DSS was followed by a two-week washout period to allow for regenerating epithelium to appear adjacent to areas of epithelial ulceration. After DSS washout, regenerating intestinal epithelium was isolated by flow cytometry, RNA was extracted, and m6A-modified transcripts were immunoprecipitated and sequenced (m6A-seq).
Results. m6A-seq yielded ~5500 total m6A-modified sites within the regenerating intestinal epithelium. This corresponded to 314 m6A-modified mRNAs that were unique to the regenerating as compared to the homeostatic epithelium. These unique transcripts were enriched for pathways involved in RNA transcription and processing, cell-cell junctions, and Hippo pathway signaling, all of which have previously been implicated in intestinal epithelial regeneration.
Conclusions. Our data support the hypothesis that m6A modifies many transcripts with known roles in intestinal epithelial regeneration. Ongoing studies are evaluating how m6A modification contributes to regulation of these transcripts and whether m6A identifies transcripts with previously unknown roles in regeneration. We are also evaluating whether m6A is required for optimal regeneration following epithelial injury.
