Investigating Ash1L Histone Methyltransferase using Chemical Biology Approaches
Notice bibliographique
Résumé
Ash1L (absent, small, or homeotic-like 1) is a histone lysine methyltransferase that catalyzes mono- and dimethylation of histone H3 lysine 36. Previous studies showed that Ash1L is implicated in a variety of cancers. Remarkably Ash1L is involved in the pathogenesis of acute leukemias with chromosomal translocations of the Mixed Lineage Leukemia (MLL) gene, which present in 5-10% of adult acute leukemias and up to 70% of infant leukemias with poor prognosis. Hence Ash1L is a validated target for therapeutic intervention, and the availability of potent small molecule inhibitors would be invaluable to investigate the roles of Ash1L in leukemias and other diseases. We previously employed fragment-based drug discovery and identified compounds that bind to Ash1L. Through optimization of the fragment hit, we developed inhibitors that exhibit sub-millimolar activity against Ash1L. In this work, we aimed to further improve the inhibitory activity of compounds and present the development of potent Ash1L inhibitors. We conducted extensive medicinal chemistry optimization and structure-based design. These efforts yielded a series of potent compounds that selectively inhibit Ash1L in biochemical assays and exhibit on-target activity in human leukemia cells. Moreover, we obtained multiple high resolution crystal structures of Ash1L in complex with inhibitors, and these structures reveal that compounds bind to a pocket in the vicinity of the autoinhibitory loop in the SET domain. To our knowledge, compounds we developed represent first-in-class inhibitory chemical probes targeting Ash1L. Based on the design of Ash1L inhibitors we made a fluorescent tool compound and developed a fluorescence polarization assay. Employing this assay we performed high-throughput screening (HTS) of 23,500 compounds and identified a hit compound that binds to Ash1L. The discovery of this compound from HTS provides a promising direction for developing a new class of Ash1L inhibitors. Lastly, we used Ash1L inhibitors to probe the conformational dynamics of Ash1L SET domain and studied the protein-protein interaction between Ash1L and Mrg15. We discovered that binding of inhibitor partially stabilizes the conformation of the SET domain but the autoinhibitory loop region experiences enhanced dynamics in solution. We also validated that Mrg15 binds to Ash1L and activates its enzymatic activity. However, we found that the activities of Ash1L inhibitors are not affected by the presence of Mrg15. In conclusion, our studies describe multiple approaches of using well-characterized chemical probes to investigate inhibition and functions of Ash1L histone methyltransferase.
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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 | 0,000 |
| 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) | 0,000 | 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 ».