Abstract 4171: Novel regulatory role of Neuropilin-1 in endothelial to mesenchymal transition as a potential source of carcinoma associated fibroblasts
Notice bibliographique
Résumé
Abstract BACKGROUND: Endothelial-mesenchymal transition (EndMT) is an extreme form of endothelial cell plasticity whereby endothelial cells acquire mesenchymal phenotype. While EndMT is a significant component during embryogenesis, maladaptive EndMT is a key contributor to fibrotic pathologies including cancer. With regard to this, EndMT accounts for up to 40% of cancer-associated fibroblasts (CAFs) that play an important role in cancer progression through potentially oncogenic signals like transforming growth factor- β (TGF-β). Recent investigations have demonstrated that the angiogenic co-receptor Neuropilin-1 (NRP1) is aberrantly expressed in cancer and modulates cancer progression through interaction with various ligands, including TGF-β. However, the relationship and driving mechanisms linking NRP1 and EndMT remain unexplored. METHODS: SiRNA-mediated NRP1 gene knockdown studies using Dharmafect-4 transfection reagent and siNRP1 or scramble control (Ambion) were performed on human umbilical vein endothelial cells (HUVECs), at baseline and after TGF-β stimulation. Total RNA was extracted using Trizol® reagent (Invitrogen), and precipitated with isopropanol. Complementary DNA was synthesized using iScript cDNA synthesis kit (Bio-Rad) and real-time polymerase chain reaction (RT-PCR) was performed using SYBR Green (Bio-Rad) following manufacturer's protocols. Markers of EndMT and mediators of TGF-β1 signalling were evaluated by RT-PCR, western blotting and immunocytochemistry. RESULTS: RT-PCR and western blots measuring NRP1 mRNA and protein levels in HUVEC lysates confirmed successful silencing. Light and fluorescence microscopy revealed marked morphological changes in TGF-β1 stimulated scramble siRNA-transfected HUVECs. TGF-β1 stimulated HUVECs demonstrated a distinct change from “cobblestone-like endothelial cell morphology” to an enlarged spindle shaped appearance consistent with a “fibroblast like morphology”, accompanied by re-arrangement of the cytoskeleton. We further observed that these characteristic EndMT features were revoked upon NRP1 silencing where cells maintained a conventional cobblestone phenotype. Expression of endothelial cell markers, CD-31 and VE-Cadherin, were significantly higher in NRP1-silenced vs. scramble siRNA-treated HUVECs; additionally, NRP1 silencing was associated with significant reduction in expression of the mesenchymal markers α-smooth muscle actin, Slug and N-Cadherin. Loss of NRP1 was accompanied by decreased expression of TGF-β1 and its receptors as well as significant changes in the key downstream effectors of TGF-β1 signalling. CONCLUSION: Our novel findings demonstrating that loss of NRP1 revokes TGF-β-dependent EndMT-like phenotype switching suggesting that NRP1 may represent a novel therapeutic target to limit the source of EndMT derived CAFs and associated cancer aggression. Citation Format: Pratiek N. Matkar, Krishna Kumar Singh, Gerald Prud'homme, Howard Leong-Poi. Novel regulatory role of Neuropilin-1 in endothelial to mesenchymal transition as a potential source of carcinoma associated fibroblasts. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4171. doi:10.1158/1538-7445.AM2015-4171
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) | 0,002 | 0,001 |
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 ».