407. A Novel Rabbit Antibody-Derived, Anti-CD123 LV/CAR Construct for AML Immunotherapy
Notice bibliographique
Résumé
Chimeric antigen receptors (CARs) have emerged in the immunotherapy field as an exciting new option for cancer treatment, with clinical trials of CD19-directed CARs having already demonstrated long-lasting responses in patients with ALL and CLL. As a hematological malignancy, acute myeloid leukemia (AML) may be another viable target for CAR-mediated therapy. Furthermore, with a 5-year survival rate of just 5.5% for patients over 65, new treatments for AML are very much needed. CD123 (the IL-3 receptor α-chain) is upregulated on AML blasts/stem cells and plays a role in proliferation and apoptotic resistance. This antigen demonstrates much lower expression levels on normal hematopoietic cells, where its expression is restricted to the myeloid progenitor subpopulation. Previous attempts to target CD123 in AML through several forms of immunotherapy have had variable success. Specifically, results of previous CD123 murine antibody-derived CARs have been mixed, with some results showing CAR-mediated eradication of normal myelopoiesis via targeting of HSCs with low CD123 expression. We have developed a CD123 CAR, derived from a novel rabbit anti-CD123 mAb that we generated. Rabbit antibodies are reported to have a broader avidity and higher range of affinities than mouse mAbs; this may lead to a CAR with a unique binding profile. We will determine whether such a rabbit-derived CD123 CAR will lead to more specific binding, resulting in optimized killing of AML cells, while minimizing cytotoxic effects on HSCs. To generate our CAR, human CD123 was purified as a GST-tagged protein and used for immunization of rabbits. Hybridoma cell lines were developed from the spleen cells of rabbits with positive immune responses, and novel antibodies were purified and screened for specificity to CD123 using a combination of ELISA, flow cytometry, and ADCC. A candidate antibody was selected, and the VL and VH chains were subcloned, sequenced, and assembled into an scFv. A second generation CAR was then designed that includes a CD8 hinge and transmembrane region, a 4-1BB costimulatory domain, and a CD3ζ signaling domain. This construct was then subcloned into a lentiviral backbone to facilitate expression in immune effector cells. Our CD123 CAR has been transduced into primary T cells and the NK-92 cell line for in vitro testing. Flow cytometry demonstrates that our CARs are expressed at the cell surface. Furthermore, the expression of the CD123 CAR has been shown to be stable in the NK-92 cell line. Cytotoxicity assays are being performed in vitro in order to confirm binding specificity and cytotoxic potential of the CD123 CARs in both NK-92 and T cells. Future work will compare CAR T and CAR NK killing in vivo using NSG mouse models of AML. AML may be an ideal target for CAR therapy, and we will exploit our novel CD123 CARs as therapeutic entities. We will examine whether the use of a novel rabbit anti-CD123 scFv in our LV/CAR construct will optimize the killing of AML cells while minimizing HSC eradication. This novel second-generation CAR has the potential to greatly impact the treatment of AML patients in the future.
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.
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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 | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,001 | 0,000 |
| Méta-épidémiologie (sens large) | 0,001 | 0,001 |
| 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) | 0,003 | 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 ».