Generation and Implementation of a Novel Murine Xenograft Model for Evaluating Human Hematopoietic Cell-Targeted Gene Therapies for Fabry Disease.
Notice bibliographique
Résumé
Abstract Abstract 3576 Poster Board III-513 Fabry disease is an X-linked lysosomal storage disorder caused by a deficiency of the enzyme α-galactosidase A (α-gal A). The inability to prevent the progression of galactosylsphingolipid deposition, such as globotriaosylceramide (Gb3), has a significant impact on quality of life and diminishes lifespan from early-onset strokes, progressive renal failure, and heart attacks. Previously, we have demonstrated that gene transfer into murine hematopoietic cells can correct the defect systemically in Fabry mice. The goal of the present study is to create a pure Fabry/NOD/SCID murine line to facilitate the in vivo assessment of human cell-targeted therapies against the disease. The pure line was generated by a “speed congenic” breeding program. The parental generation (F0) was represented by a C3H+C57BL/6 Fabry female mouse (α-gal A−/− scid+/+) and a NOD/SCID male mouse (α-gal A+/0 scid−/−). To generate the α-gal A-/+ scid−/− female mice (F2), the double heterozygous female mice from F1 were mated with NOD/SCID male mice. F3 and all the subsequent generations (until F11) were derived by backcrossing α-gal A-/+ scid−/− female mice with α-gal A+/0 scid−/− male mice. At this point, genome scanning analysis, fluorometric enzymatic assays, and HPLC assessment revealed that the F11 purity was higher than 99%; α-gal A activity was reduced significantly in plasma and Gb3 levels were increased considerably in heart, liver, spleen, kidney and lung, in comparison to NOD/SCID control mice. With the aim of obtaining the pure Fabry/NOD/SCID line, F11 mice were crossed with each other (α-gal A-/+ scid−/− female mice with α-gal A-/0 scid−/− male mice) and then F12 double homozygous female mice (α-gal A−/− scid−/−) were crossed with F12 hemizygous (α-gal A-/0 scid−/−) male mice. For each generation, the genotype of offspring was analyzed by PCR and the absence of T and B cells was confirmed phenotypically by flow cytometry. Currently, this new xenograft model is being validated by using hematopoietic cell targets. To this end, normal human mobilized CD34+ cells were separately transduced with a control (eGFP lentivector) or a bicistronic lentiviral vector encoding the human α-gal A and the human CD25 cell surface marker. 8×105 cells were injected intravenously into sub-lethally irradiated 8-week-old pure Fabry/NOD/SCID male mice. Injected cells were 25% positive for eGFP and 34% positive for human CD25 expression, respectively. Presence of human CD45+ cells, CD45+/CD25+ cells, CD45+/eGFP+ cells and α-gal A activity will be regularly monitored on peripheral blood or plasma, respectively. Near future studies will include Gb3 quantification in tissues at sacrifice and secondary recipient transplantation. As well, Fabry patient bone marrow cells are currently being collected under an approved protocol for testing in this model. In conclusion, this novel xenograft Fabry model is a key tool for developing different therapies for Fabry Disease and will help us to reduce the gap between the bench and the clinic. Disclosures: No relevant conflicts of interest to declare.
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 | 0,001 | 0,000 |
| Méta-épidémiologie (sens strict) | 0,001 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,001 |
| Bibliométrie | 0,001 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,001 | 0,001 |
| Science ouverte | 0,001 | 0,000 |
| Intégrité de la recherche | 0,001 | 0,001 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,003 | 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 ».