Osencabtagene Autoleucel

autologous T lymphocytes obtained from peripheral blood by leukapheresis, transduced with two self-inactivating, nonreplicating lentiviral vectors encoding: (i) a chimeric antigen receptor (CAR) targeting human CD7: the expressed transgene comprises a granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor signal sequence, a single-domain antibody derived from a humanized nanoantibody (HuVHH6) targeting CD7, an immunoglobulin G4 (IgG4) Fc hinge, an inducible T lymphocyte costimulator (ICOS) transmembrane and intracellular domain, a 4-1BB intracellular costimulatory domain and a CD3ζ intracellular activation domain, under control of the human elongation factor 1 alpha (EF1α) promoter. (ii) a CD7-blocking gene comprising a granulocyte-macrophage colonystimulating factor (GM-CSF) receptor signal sequence, a bivalent CD7 nano-antibody sequence (VHH6-linker-VHH6) fused to an endoplasmic reticulum (ER) retention sequence (KDEL sequence), under control of the cytomegalovirus promoter. In both constructs, the transgene is flanked by 5' and 3' long terminal repeats (LTRs) and also contains a ψ packaging signal, a Rev response element (RRE) and a central polypurine tract (cPPT) sequence 5' to the transgene, and a Woodchuck hepatitis virus post-transcriptional regulatory element (WPRE)3' to the transgene. Both vectors are pseudotyped with vesicular stomatitis virus (VSV) G glycoprotein. The leukapheresis material is enriched for CD4+ and CD8+ T lymphocytes by positive immunoselection prior to activation with CD3 and CD28 agonists in growth media containing interleukin 7 (IL-7) and interleukin 15 (IL-15). The cells are then transduced with the lentiviral vectors and expanded in growth media containing IL-7 and IL-15. The cell suspension consists of CD3+ T lymphocytes (≥90%; of which CD3+CD7- T lymphocytes account for more than 95%), with greater than 5% of the T lymphocytes expressing the CAR transgene

AUTOLOGOUS T LYMPHOCYTES OBTAINED FROM PERIPHERAL BLOOD BY LEUKAPHERESIS, TRANSDUCED WITH TWO SELF-INACTIVATING, NONREPLICATING LENTIVIRAL VECTORS ENCODING: (I) A CHIMERIC ANTIGEN RECEPTOR (CAR) TARGETING HUMAN CD7: THE EXPRESSED TRANSGENE COMPRISES A GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR (GM-CSF) RECEPTOR SIGNAL SEQUENCE, A SINGLE-DOMAIN ANTIBODY DERIVED FROM A HUMANIZED NANOANTIBODY (HUVHH6) TARGETING CD7, AN IMMUNOGLOBULIN G4 (IGG4) FC HINGE, AN INDUCIBLE T LYMPHOCYTE COSTIMULATOR (ICOS) TRANSMEMBRANE AND INTRACELLULAR DOMAIN, A 4-1BB INTRACELLULAR COSTIMULATORY DOMAIN AND A CD3.ZETA. INTRACELLULAR ACTIVATION DOMAIN, UNDER CONTROL OF THE HUMAN ELONGATION FACTOR 1 ALPHA (EF1.ALPHA.) PROMOTER. (II) A CD7-BLOCKING GENE COMPRISING A GRANULOCYTE-MACROPHAGE COLONYSTIMULATING FACTOR (GM-CSF) RECEPTOR SIGNAL SEQUENCE, A BIVALENT CD7 NANO-ANTIBODY SEQUENCE (VHH6-LINKER-VHH6) FUSED TO AN ENDOPLASMIC RETICULUM (ER) RETENTION SEQUENCE (KDEL SEQUENCE), UNDER CONTROL OF THE CYTOMEGALOVIRUS PROMOTER. IN BOTH CONSTRUCTS, THE TRANSGENE IS FLANKED BY 5' AND 3' LONG TERMINAL REPEATS (LTRS) AND ALSO CONTAINS A .PSI. PACKAGING SIGNAL, A REV RESPONSE ELEMENT (RRE) AND A CENTRAL POLYPURINE TRACT (CPPT) SEQUENCE 5' TO THE TRANSGENE, AND A WOODCHUCK HEPATITIS VIRUS POST-TRANSCRIPTIONAL REGULATORY ELEMENT (WPRE)3' TO THE TRANSGENE. BOTH VECTORS ARE PSEUDOTYPED WITH VESICULAR STOMATITIS VIRUS (VSV) G GLYCOPROTEIN. THE LEUKAPHERESIS MATERIAL IS ENRICHED FOR CD4+ AND CD8+ T LYMPHOCYTES BY POSITIVE IMMUNOSELECTION PRIOR TO ACTIVATION WITH CD3 AND CD28 AGONISTS IN GROWTH MEDIA CONTAINING INTERLEUKIN 7 (IL-7) AND INTERLEUKIN 15 (IL-15). THE CELLS ARE THEN TRANSDUCED WITH THE LENTIVIRAL VECTORS AND EXPANDED IN GROWTH MEDIA CONTAINING IL-7 AND IL-15. THE CELL SUSPENSION CONSISTS OF CD3+ T LYMPHOCYTES (>=90%; OF WHICH CD3+CD7- T LYMPHOCYTES ACCOUNT FOR MORE THAN 95%), WITH GREATER THAN 5% OF THE T LYMPHOCYTES EXPRESSING THE CAR TRANSGENE