A155 THE IMMUNOMODULATOR, LEFLUNOMIDE, REDUCES MITOCHONDRIAL FRAGMENTATION CAUSED BY ADHERENT–INVASIVE E. COLI AND PERSISTENCE OF THE PATHOBIONT WITHIN ENTERIC EPITHELIA
Bibliographic record
Abstract
Mitochondria can exist as a dynamic network that constantly remodels via the processes of fission and fusion (mitochondrial dynamics) to meet cellular energy demands and allow recycling of damaged mitochondria. Mitochondrial dysfunction can occur in inflammatory bowel disease. Adherent-invasive E. coli (AIEC) are a putative etiological agent in Crohn’s disease. It is unknown if infection with AIEC affects mitochondrial dynamics in intestinal epithelial cells. It was recently discovered that leflunomide, an FDA approved anti-inflammatory and antirheumatic drug, promotes mitochondrial fusion. (1) Explore the relationship between mitochondrial dynamics and epithelial function in vitro and determine if, then how, infection with AIEC affects mitochondrial dynamics and any consequence for permeability. (2) Determine if leflunomide blocks any of the effects of AIEC on epithelial function. Human colon-derived epithelial lines were cultured with non-invasive E. coli as a control, viable AIEC (LF82: 4h 108 CFU/mL or 16h 104 CFU/mL), fixed (dead) AIEC, spent medium from AIEC cultures ± co-treatment with inhibitors of mitochondrial fission (i.e. P110, Mdivi1) or leflunomide. Epithelia were examined: (a) electron microscopy; (b) ATP; (c) live-cell imaging of mitochondria morphology and membrane potential; (d) immunoblotting for Dynamin-Related Peptide-1 (Drp1); (e) barrier function; (f) cytochrome c release from mitochondria, and (g) intracellular AIEC. Infection with AIEC resulted in reduced mitochondrial membrane potential and ATP levels, and dramatic Drp1-dependent mitochondrial fragmentation. Fragmentation was dependent on viable AIEC with the ability to adhere and invade epithelia. While P110, Mdivi1 and leflunomide all acutely (2-4h) prevented AIEC-induced mitochondrial fragmentation, this was eventually overcome, such that AIEC-induced loss of the epithelial barrier function and apoptosis eventually emerged (~8h post-treatment with 108 CFU). Leflunomide significantly reduced numbers of AIEC inside cells. Mitochondrial fragmentation is identified as a novel aspect of AIEC-epithelial interaction that will contribute to loss of function. Precise understanding of the molecular process that facilitate the mitochondrial fragmentation could open new therapeutic avenues relevant to inflammatory disease and/or that due to microbial pathogens. Finally, by virtue of its ability to reduce the intracellular burden of AIEC (possibly linked to its capacity to promote mitochondrial fusion), leflunomide could be a treatment for IBD in which AIEC are implicated. CIHRNSERC, UofC Eyes High, Izaak-Killam
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 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 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
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 teacher head, 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".