A41 ENDOPLASMIC RETICULUM STRESSED HUMAN COLON ORGANOIDS CONDITION DENDRITIC CELLS TO A PRO-INFLAMMATORY PHENOTYPE
Bibliographic record
Abstract
Endoplasmic reticulum (ER) stress occurs when cellular protein modification and secretion are perturbed from accumulation of misfolded or improperly modified proteins. ER stress leads to activation of a cellular protection pathway, the unfolded protein response (UPR), to restore homeostasis. Recent studies showed that ER stress and an overactive UPR in intestinal epithelial cells (IECs) predispose to murine colitis and may underlie human IBD. We reported that two stress signals, ATP and H2O2, alter IEC toll-like receptor (TLR) signaling, and that stressed IECs condition dendritic cells (DCs) to a pro-inflammatory state. Here we aimed to assess whether extracellular (ATP) or intracellular (Tg) stress alter immune responses to FliC in human colonic organoids, and how these stressed organoids affect DC maturation. Ascending colon mucosal biopsies were differentiated into enteroids, split into monolayers, and grown for 5–7 days until confluent and polarized. Cells were stressed with 100nM Tg overnight, 100uM of ATP for 20 mins, or vehicle control; media was changed, and 1ug/ml of FliC added. 1h later, media was changed to remove residual FliC, and supernatants were after 2h. MoDCs (100,000/well) were incubated with 100ul of conditioned supernatants for 24 hours, cells were harvested, and receptor expression measured by flow cytometry. TLR5 stimulation with flagellin increased IL-8 production from organoids, and this was significantly higher in organoids pretreated with Tg (P= 0.0109). Supernatants from Organoids treated with FliC plus Tg or ATP conditioned moDCs to increase surface expression of MHCII and costimulatory molecules CD86 and CD80, and decrease expression of CD103, a tolerogenic marker. Pearson correlation analysis between the fold-change of IL-8 produced by organoids stimulated with FliC + Tg alone, compared to CD86 or CD80 upregulation revealed a statistically significant association between stress responses in the organoid IECs and DC CD80 and CD86 expression (p < 0.05 for each), suggesting that DC costimulatory marker overexpression is dependent on the presence of ATP- or Tg-induced stress more than TLR5 stimulation. Moreover, organoid gene expression of Relmβ and TSLP was increased (Relmβ: Tg 2.14, Tg+FliC 2.38; TSLP: Tg 3.82, and Tg+FliC 3.37-fold increase vs unstimulated organoids, respectively). This suggests that IL-8, Relmβ, and/or TSLP could be responsible for the maturation and pro-inflammatory shift in moDCS. We found that colonic organoids increase inflammatory responses to flagellin in the presence of ATP or ER stress, and that supernatiants from these cells condition DCs to an inflammatory surface marker phenotype. Together these findings suggest a link between the UPR in IECs and the damaging T cell responses that characterize inflammatory bowel disease. CCC, CIHR
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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) | 0.005 | 0.002 |
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".