A59 GLOBAL PROTEOMIC PROFILING OF HUMAN COLONOID MONOLAYERS UNDERGOING IN VITRO CHRONIC DAMAGE
Notice bibliographique
Résumé
Abstract Background An in vitro damage model has been established in our lab using human colonoids grown as 2D monolayers. Upon being subjected to repeated rounds of air-liquid interface (ALI) growth and injury by submergence, these colonoid monolayers lost their barrier integrity and regrowth potential. Changes in mRNA expression and DNA methylation in genes from this human model of injury were similar to those that occur in Inflammatory Bowel Disease (IBD) and colon cancer. Significant morphological changes were observed in these monolayers after they were subjected to subsequent rounds of submergence injury, compared to when they were differentiated in ALI. Purpose Submergence injury is predicted to be involved in unfolded protein response (UPR) activation which can specifically alter translation. Hence proteomics studies will help undertand these changes. Method To determine if these changes are mirrored in the proteomes of damaged colonoids, we employed a Single-Plot, Solid-Phase-enhanced Sample Preparation (SP3) technology for Mass Spectrometry (MS) based proteomics analysis to characterize these monolayers at baseline, once they were differentiated in ALI, after one and five rounds of injury after differentiation in ALI, and after stimulation with the Toll-like receptor 5 (TLR5) agonist FliC. Hierarchical clustering, enrichment analysis, volcano plot analysis after pre-processing and normalization of the proteomics data set revealed differentially expressed proteins across various groups of monolayers. Result(s) Preliminary proteomic data analysis revealed changes in the profile of proteins involved in cellular differentiation, mitochondrial proteins, hypoxia upregulated proteins, those responsible for the maintenance and reorganization of the cytoskeletal structure and Golgi structure. These changes in protein profile may account for the significant morphological changes observed in these monolayers when subjected to submergence injury. Some outliers in monolayers subjected to microbial stimulation included proteins involved in regulation of extracellular matrix dependent motility and components of Adaptor Protein Complexes. Further studies are needed to ascertain if these account for the protective effect of FliC on these monolayers. Conclusion(s) This study suggests that the submergence injury to these healthy human derived colonoid monolayers leads to changes in their protein profile which mirror those seen in case of acute and chronic inflammation like IBD and colon cancer. It corroborates with the findings of gene expression and epigenetic analyses using the in vitro model established in our lab. Please acknowledge all funding agencies by checking the applicable boxes below CCC Disclosure of Interest None Declared
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 ».