A21 OVEREXPRESSION OF <i>ASCL2</i> ALTERS DIFFERENTIATION IN ESOPHAGEAL ORGANOIDS
Bibliographic record
Abstract
Abstract Background The first population of stem cells in the esophageal epithelium was recently identified with the marker Keratin 15 ( Krt15). However, little is known about the mechanisms underlying the expansion and the function of these stem cells. It was shown that the transcription factor ASCL2 is upregulated in Krt15+ cells compared to Krt15- cells. Interestingly, ASCL2 is a gene target of the Wnt/β-catenin pathway, which acts as a regulator of proliferation and maintenance of the stemness state. The ultimate goal of my research project is to determine the role of ASCL2 in the maintenance of esophageal stem cells and to identify its binding partners. Aims Investigate the role of ASCL2 in esophageal epithelial biology. Methods Lentiviral infection approach was used to obtain mouse esophageal organoids overexpressing ASCL2. Organoid culture, immunostaining (such as IF and H&E), qPCR, WB and proliferation assay were used to characterize the effect of ASCL2 overexpression on morphology, differentiation, proliferation, self-renewal and gene expression. Results First, ASCL2 overexpression was confirmed by WB. Interestingly, the morphology of organoid overexpressing ASCL2 was severely altered: organoids were smaller and less differentiated. Defect in differentiation was investigated by qPCR and IF using relevant markers such as p63, Krt13, Wnt5a and NT5E. Indeed, we observed an increase in basal marker ( p63), a decrease in suprabasal markers ( Krt13, Wnt5a) and in a stem cell marker ( NT5E). We also investigated the role of ASCL2 in self-renewal and observed that organoid formation capacity was reduced in ASCL2-overexpressing organoids. Furthermore, proliferation was also reduced in WST-1 assays. We also observed lower expression of the gene Top2a, a recently identified marker of the proliferative basal cell population in the human esophagus. Finally, we observed significant changes in the expression of genes associated with quiescent stem cells (Clu, ZFP36L2 and Anxa1). Conclusions ASCL2 overexpression alters differentiation and proliferation in organoids. ASCL2 could play a role in orchestrating cell fate decision in the esophageal epithelium. Funding Agencies NSERC, Canada Research Chair
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.001 |
| Insufficient payload (model declined to judge) | 0.002 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".