Abstract 84: nAG (a Salamander-Derived Protein) as an Inhibitor of TGF-β Signaling and Fibrotic Responses
Notice bibliographique
Résumé
PURPOSE: Salamanders have the amazing ability to regenerate their limbs within 30 days when amputated. The key protein responsible for this regeneration is the nAG protein, which stands for newt anterior gradient. In previous studies, a nAG gene was designed that was suitable for human use that demonstrated local injection of recombinant nAG protein reduces hypertrophic scarring in a rabbit ear model. Fibrotic disorders of the skin such as scleroderma, hypertrophic scarring and keloids are characterized by excessive TGF-β, leading to an increase in deposition of collagen and other extracellular matrix (ECM) components, resulting in functional impairment which is often debilitating and treatment options remain limited. The aim of this study is to assess the effect nAG has on the fibrotic process of human scleroderma fibroblasts. METHODS: Fibroblasts from lesional scleroderma patient skin were treated with nAG protein in doses of 100pM,1nM and 10nM for 24 hrs and were then left untreated or treated with 20 pM of TGF-β. The inhibition of TGF-β-mediated pro-fibrotic responses was determined by measuring the Extracellular Matrix (collagen III, Fibronectin), connective tissue growth factor (CTGF) and myofibroblast alpha smooth muscle actin (α-SMA) protein production by Western blot as well as immunofluorescence and validated at the mRNA level by Quantitative PCR. Activation of the TGF-β pathway was determined by measuring the TGF-β receptor 1 (ALK5) and phosphorylated Smad2/3 levels by using Western blot and immunofluorescence. Luciferase assay was used to measure nAG protein’s capacity to inhibit the thee TGF-β isomers. Cell migration was assessed using a scratch assay and finally colocalization of nAG protein with TGF- β receptor 1 was evaluated using confocal microscopy. RESULTS: Both the Western Blot and the immunofluorescence results revealed that the application of the nAG protein to human scleroderma fibroblasts in the presence of TGF-β successfully inhibited the fibrotic response shown by a decrease in the fibrotic factors such as collagen III, alpha smooth muscle actin (α-SMA), connective tissue growth factor (CTGF) and fibronectin. In addition, immunofluorescence and western blot after 1 hour of treatment with nAG revealed a significant decrease in Phosphorylated Smad2/3, and a decrease in the TGF-β receptor 1 (ALK5) inhibiting the TGF- β pathway. Luciferase assay revealed nAG’s inhibition of the canonical TGF-β pathway to be most specific with TGF-β1 isomer reducing activity by 83%. Cell migration was significantly inhibited with nAG protein treatment and confocal microscopy revealed the colocalization of nAG protein with ALK5 receptors. CONCLUSION: Fibrosis in scleroderma fibroblasts was effectively inhibited when treated with nAG protein, demonstrated by the decrease in ECM using Western Blot and immunofluorescence. Although much about the mechanism of the nAG protein is still unknown, the decrease in pSmad2/3 and ALK5 receptors most potently with TGF-β1 after treatment, inhibition of cell migration and binding of nAG protein with ALK5 receptors suggest that nAG blocks the canonical TGF-β pathway. This research is anticipated to lead to the development of an injectable antifibrotic agent for the treatment of fibrotic disorders such as scleroderma, hypertrophic scars and keloids. A. Al-Qattan: None. L. Lessard: None. A. Philip: None.
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 distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,000 | 0,002 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,001 |
| 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 tête enseignante, 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 ».