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Updated: Oct 24, 2025
In vitro Organoid Culture of Primary Mouse Colon Tumors
Published on: May 17, 2013
Transcriptome-wide In Vitro Effects of Aspirin on Patient-derived Normal Colon Organoids
Matthew A M Devall1, David A Drew2,3, Christopher H Dampier1
1Center for Public Health Genomics, Department of Public Health Sciences, University of Virginia, Charlottesville, Virginia.
Abstract:
Mechanisms underlying aspirin chemoprevention of colorectal cancer remain unclear. Prior studies have been limited because of the inability of preclinical models to recapitulate human normal colon epithelium or cellular heterogeneity present in mucosal biopsies. To overcome some of these obstacles, we performed in vitro aspirin treatment of colon organoids derived from normal mucosal biopsies to reveal transcriptional networks relevant to aspirin chemoprevention. Colon organoids derived from 38 healthy individuals undergoing endoscopy were treated with 50 μmol/L aspirin or vehicle control for 72 hours and subjected to bulk RNA sequencing. Paired regression analysis using DESeq2 identified differentially expressed genes (DEG) associated with aspirin treatment. Cellular composition was determined using CIBERSORTx. Aspirin treatment was associated with 1,154 significant (q < 0.10) DEGs prior to deconvolution. We provide replication of these findings in an independent population-based RNA-sequencing dataset of mucosal biopsies (BarcUVa-Seq), where a significant enrichment for overlap of DEGs was observed (P < 2.2E-16). Single-cell deconvolution revealed changes in cell composition, including a decrease in transit-amplifying cells following aspirin treatment (P = 0.01). Following deconvolution, DEGs included novel putative targets for aspirin such as TRABD2A (q = 0.055), a negative regulator of Wnt signaling. Weighted gene co-expression network analysis identified 12 significant modules, including two that contained hubs for EGFR and PTGES2, the latter being previously implicated in aspirin chemoprevention. In summary, aspirin treatment of patient-derived colon organoids using physiologically relevant doses resulted in transcriptome-wide changes that reveal altered cell composition and improved understanding of transcriptional pathways, providing novel insight into its chemopreventive properties. PREVENTION RELEVANCE: Numerous studies have highlighted a role for aspirin in colorectal cancer chemoprevention, though the mechanisms driving this association remain unclear. We addressed this by showing that aspirin treatment of normal colon organoids diminished the transit-amplifying cell population, inhibited prostaglandin synthesis, and dysregulated expression of novel genes implicated in colon tumorigenesis.
Insights
Aspirin treatment of colon organoids reduced transit-amplifying cells and altered gene expression, offering new insights into colorectal cancer chemoprevention mechanisms. This study reveals key pathways involved in aspirin's preventive effects.
Area of Science:
- Gastroenterology and Hepatology
- Oncology
- Molecular Biology
Background:
- Aspirin is recognized for colorectal cancer chemoprevention, but its underlying mechanisms are not fully understood.
- Previous research faced limitations due to preclinical models' inability to replicate human colon epithelium and cellular heterogeneity.
Purpose of the Study:
- To investigate the transcriptional networks involved in aspirin's chemopreventive effects on colorectal cancer using patient-derived colon organoids.
- To identify novel molecular targets and pathways modulated by aspirin in normal colon epithelium.
Main Methods:
- Colon organoids from 38 healthy individuals were treated with aspirin (50 μmol/L) or vehicle control.
- Bulk RNA sequencing and differential gene expression analysis (DESeq2) were performed.
- Cellular composition was analyzed using single-cell deconvolution (CIBERSORTx), and gene co-expression networks were constructed.
Main Results:
- Aspirin treatment resulted in 1,154 differentially expressed genes (DEGs) and a significant decrease in transit-amplifying cells.
- Replication of findings was confirmed in an independent dataset (BarcUVa-Seq), showing significant overlap in DEGs.
- Novel targets like TRABD2A and pathways involving EGFR and PTGES2 were identified, with PTGES2 previously linked to aspirin chemoprevention.
Conclusions:
- Aspirin treatment of patient-derived colon organoids reveals transcriptome-wide changes and altered cell composition, enhancing understanding of its chemopreventive properties.
- Findings suggest aspirin diminishes transit-amplifying cells, inhibits prostaglandin synthesis, and affects novel genes crucial in colon tumorigenesis, providing new avenues for chemoprevention research.
