79. Development and Validation of a Globally Portable Platform for Lentivirus Mediated Hematopoietic Stem Cell Gene Therapy
Notice bibliographique
Résumé
Lentivirus (LV) mediated gene therapy of CD34+ hematopoietic stem and progenitor cells (HSPCs) has demonstrated clinical success for a variety of diseases. However, current state-of-the-art requires ex vivo HSPC gene transfer in a dedicated Good Manufacturing Practices (GMP) facility, limiting treatment to highly developed countries capable of supporting GMP infrastructure. We developed a flexible, overnight platform for efficient isolation and LV gene modification of bone marrow and mobilized peripheral blood CD34+ HSPCs in a closed, table top system, the Prodigy CliniMACS™, with limited requirement for additional equipment. We performed all experiments with only a biosafety cabinet and personal protective equipment to simulate anticipated conditions in clinical facilities of underdeveloped countries. Given the economic burden of mobilization, we initially designed the process for bone marrow. A total of 7 custom programs were developed based on current device memory limitations: (1) hetastarch sedimentation to deplete red blood cells (RBCs), (2) labeling CD34+ cells in the RBC-depleted product, (3) immunomagnetic enrichment of CD34+ cells, (4) initial transduction (MOI = 20 IU/cell), (5) culture overnight, (6) second transduction (MOI = 20 IU/cell) and additional culture, and (7) harvest and formulation of the final product. Addition of a pyrimidoindole derivative, UM729, permitted efficient transduction of CD34+ HSPCs in £18 hours. The process took <36 hours from bone marrow collection to infusion of the gene modified cell product, with £3 hours hands-on time. Autologous, LV gene-modified CD34+ HSPCs from two nonhuman primates produced using this platform engrafted and supported multilineage hematopoietic repopulation after myeloablative total body irradiation. Total cell doses achieved were 27 × 107 and 5.4 × 107 CD34+ cells/kg body weight, respectively. Transduction efficiency in colony-forming units was 23% and 39%, respectively. We observed stable, persistent gene marking in peripheral blood cells up to 16% within one month after infusion. Neither animal displayed evidence for increased toxicity, including potential contamination from the cell product. We then validated processing of human mobilized apheresis products. We demonstrate efficient isolation of human CD34+ HSPCs and up to 60% transduction efficiency with a clinical-grade LV currently being tested in a phase I clinical trial for treatment of HIV-associated lymphoma. Immediate release criteria for product infusion were met (negative gram stain, endotoxin <5 EU/mL and cell viability >70%). Perform-and-report testing and xenotransplantation of these gene-modified cells into immunodeficient mice for further functional testing are in progress. These data demonstrate preclinical safety and feasibility of this portable strategy for ex vivo LV gene transfer into HSPCs, representing the first globally applicable advance in translation of HSPC gene therapy.
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 | 0,001 | 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 ».