Воламкабтаген дурзигедлейцел

Rec. INN (наименование, зарегистрированное ВОЗ)

allogeneic T cells obtained from peripheral blood by leukapheresis, genetically modified by CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) mediated gene editing consisting of three guide RNAs (gRNAs) introduced transiently as Cas9-gRNA ribonucleoprotein (RNP) complex for the targeted disruption of the T cell receptor alpha chain constant (TRAC), β₂ microglobulin (B2M) and the cluster of differentiation 70 (CD70) loci and insertion of an anti-CD70 chimeric antigen receptor (CAR) transgene into the TRAC locus via an adeno-associated virus serotype 6 (AAV6) vector. The CAR is composed of a signal peptide sequence from human CD8a, an anti-CD70 single-chain variable fragment (scFv) derived from anti-CD70 hybridoma 1F6, a CD8 hinge and transmembrane domain, and the 4-1BB and CD3ζ co-stimulatory domains. Expression of the CAR is driven by the elongation factor 1 alpha (EF-1α) promoter and is terminated by a synthetic polyadenylation (polyA) sequence. The anti-CD70 CAR expression cassette is flanked by two TRAC homology arms guiding the expression cassette to the TRAC locus.
The leukopheresis material is enriched for T cells using CD4/CD8 selection and activated with anti-CD3/CD28 antibody-coated beads. T cells are cultured in the presence of growth media containing human serum, interleukin 2 (IL-2) and 7 (IL-7). The final T cells are ≥30% CAR+ T cells, ≤0.4% TCR+, ≤2% CD70+, and ≤30% B2M+ and secrete interferon gamma (IFNγ) and IL-2 when co-cultured with CD70 expressing cells.

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