Production of Bioactive Peptides by Neutrophil Elastase Cleavage of E‐cadherin
Notice bibliographique
Résumé
BACKGROUND The inflammatory microenvironment in the gut contains a variety of proteases from numerous sources including inflammatory cells. We have been studying the ability of proteases to induce a switch in the colonic epithelium from a barrier to a repair phenotype (epithelial to mesenchymal transition [EMT]) characterized by increased migration and disrupted homeostasis. EMT involves the degradation of junctional proteins such as E‐cadherin [Ecad], but the functional consequences of Ecad loss remain incompletely understood. AIM To test the hypothesis that inflammatory proteases cleave Ecad to produce peptides that alter epithelial homeostasis by altering cell migration, proliferation, or cell death. METHODS Recombinant Ecad was incubated with neutrophil elastase [NE] in a minimized cell‐free system and 24 peptides identified by mass spectrometry were chosen for high‐throughput screening based on frequency, p‐value scoring, and accessibility of peptide cleavage site. Synthesized peptides were tested for biological activities including proliferation, cell death, and cell migration. Murine CMT‐93 intestinal epithelial cells were cultured in 96 well plates and wounded 5 days post‐confluency using the EssenBio WoundMaker™ tool. Plates were imaged with the IncuCyte™ live‐cell imaging system for 24–48 hours following exposure to 1, 10, and 100 mg/mL concentrations of peptides. Cells were transfected with a GFP marker for automated cell counting to measure proliferation. Cytotoxicity was determined using Cytotox dye entering cells undergoing membrane degradation. All analysis was done using the IncuCyte™ ZOOM platform in conjunction with ImageJ software. RESULTS Basolateral stimulation of human Caco2 and murine CMT93 cells with NE confirmed the ability of NE to produce C‐ and N‐terminal Ecad fragments in vitro. From our synthesized peptides, we identified 10 Ecad peptides that significantly influenced wound closure rates, both positively and negatively. Two peptides (100 mg/mL) significantly inhibited wound healing rate by 13–26% compared to vehicle controls. Six peptides (100 μg/mL) significantly increased wound healing rates by 7–11% and three peptides (10 μg/mL) significantly increased wound healing rates by 6–13% compared to vehicle controls. Preliminary results suggest that several of these peptides also have cytostatic or pro‐proliferative activity. CONCLUSIONS Our results suggest that degradation of cell junction proteins by inflammatory proteases can create bioactive peptides that alter epithelial homeostasis and alter cell migration. Our data reveal a novel and complex pathway whereby epithelia respond to inflammatory tissue damage at the cellular level to alter a barrier phenotype to a more dynamic epithelium. Support or Funding Information Funding: Supported by the Canadian Institutes of Health Research (CIHR)
Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.
Comment cette classification a été obtenuedéplier
Prédiction machine sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Le volet Gemma est une étiquette directe du modèle pour chaque travail de la base, lue sur la notice réduite au titre. Le volet Codex est un classifieur appris des 10 348 étiquettes directes de Codex et calibré sur les taux pondérés de l'échantillon; les champs sans appui suffisant ne portent aucun appel Codex. Le mode candidate est l'union des deux volets; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont pas des étiquettes humaines.
Scores du classifieur distillé par catégorie (deux têtes)
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,001 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule source (Gemma direct ou Codex distillé), pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».