Abstract 6588: Characterization of TGFb1-induced ApoEVs and their potential role in epithelial cell plasticity
Notice bibliographique
Résumé
Abstract Extracellular vesicles (EVs) are membranous particles that carry cargo such as proteins, lipids, and nucleic acids that can be crucial for intercellular communication. Epithelial-mesenchymal transition (EMT) is implicated in tissue remodeling and has been documented in the mammary gland during involution. Transgenic models and clinical studies suggest that both post-lactational and age-related lobular involution may increase susceptibility for breast cancer development. Transforming growth factor-beta 1 (TGFβ1) induces apoptosis of epithelial cells during involution and is also a potent inducer of EMT. TGFβ-induced apoptosis generates a subpopulation of EVs called apoptotic cell-derived EVs (ApoEVs), a potential cell-cell communication avenue that may be crucial for tissue remodeling. Here, we optimized a protocol to generate, isolate, and analyze TGFβ-induced ApoEVs. This may reveal potential EV-mediated signaling in driving EMT, tissue remodeling and cancer susceptibility in the mammary gland. The murine mammary gland epithelial cell line NMuMG was used, as they are known to both die and undergo EMT in response to TGFβ1 stimulation. NMuMG cells were cultured in DMEM with 10% FBS, 10 µg/mL insulin, and 1% antibiotics. Once 90% confluency was reached, growth media was replaced with starvation media consisting of DMEM, 5% EV-depleted FBS, and antibiotics. After 2.5 hours, one plate was treated with 5 ng/mL of TGFβ1 while the other received PBS. The conditioned media (CM) produced was centrifuged at 400g for 20 minutes to remove debris and then ultracentrifuged for 1 hour at 4ºC at a speed of 10, 000g to generate a pellet of large EVs. This was followed by two PBS washes under the same ultracentrifugation conditions. Next, the CM was again ultracentrifuged at a speed of 100, 000g to isolate smaller EVs. Both EV populations were resuspended in 10mM Hepes (pH 7.2) and characterized by western blot, dynamic light scattering (DLS), and transmission electron microscopy (TEM). Western blots for markers of EVs and potential contaminants suggest successful EV isolation. Blotting for apoptotic markers presented results consistent with ApoEVs, which were enriched in the samples obtained by 10, 000g ultracentrifugation. DLS analysis showed that EVs released by TGFβ-treated cells were larger and more abundant compared to those released by untreated cells; TEM analysis confirmed these results. Culturing NMuMG cells with either full CM or EV-depleted CM from either control or TGFβ1-treated cells showed morphological changes consistent with cell plasticity in response to EVs released by TGFβ1-treated cells. Our findings suggest that EVs produced during TGFβ1-induced apoptosis of NMuMG cells contain cargo capable of modulating epithelial phenotype and may aid directly or indirectly in tissue remodeling. Additional biological studies on target cells are necessary to confirm our results. Citation Format: Daxime F. Génier, Sarai A. Gomez Perez, Alicia Viloria-Petit. Characterization of TGFb1-induced ApoEVs and their potential role in epithelial cell plasticity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6588.
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 ».