title: Lentivirus-induced 'Smart' dendritic cells: Pharmacodynamics and GMP-compliant production for immunotherapy against TRP2-positive melanoma.
abstract: Monocyte-derived conventional dendritic cells (ConvDCs) loaded with melanoma antigens showed modest responses in clinical trials. Efficacy studies were hampered by difficulties in ConvDC manufacturing and low potency. Overcoming these issues, we demonstrated higher potency of lentiviral vector (LV)-programmed DCs. Monocytes were directly induced to self-differentiate into DCs (SmartDC-TRP2) upon transduction with a tricistronic LV encoding for cytokines (granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4)) and a melanoma antigen (tyrosinase-related protein 2 (TRP2)). Here, SmartDC-TRP2 generated with monocytes from five advanced melanoma patients were tested in autologous DC:T cell stimulation assays, validating the activation of functional TRP2-specific cytotoxic T lymphocytes (CTLs) for all patients. We described methods compliant to good manufacturing practices (GMP) to produce LV and SmartDC-TRP2. Feasibility of monocyte transduction in a bag system and cryopreservation following a 24-h standard operating procedure were achieved. After thawing, 50% of the initial monocyte input was recovered and SmartDC-TRP2 self-differentiated in vitro, showing uniform expression of DC markers, detectable LV copies and a polyclonal LV integration pattern not biased to oncogenic loci. GMP-grade SmartDC-TRP2 expanded TRP2-specific autologous CTLs in vitro. These results demonstrated a simpler GMP-compliant method of manufacturing an effective individualized DC vaccine. Such DC vaccine, when in combination with checkpoint inhibition therapies, might provide higher specificity against melanoma.
authors.affiliation: Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
authors.forename: B S
authors.initials: BS
authors.lastname: Sundarasetty
authors.affiliation: Department of Hematological Medicine, Cell and Gene Therapy at King's, The Rayne Institute, King's College London, London, UK.
authors.forename: L
authors.initials: L
authors.lastname: Chan
authors.affiliation: Department of Hematological Medicine, Cell and Gene Therapy at King's, The Rayne Institute, King's College London, London, UK.
authors.forename: D
authors.initials: D
authors.lastname: Darling
authors.affiliation: Department of Hematological Medicine, Cell and Gene Therapy at King's, The Rayne Institute, King's College London, London, UK.
authors.forename: G
authors.initials: G
authors.lastname: Giunti
authors.affiliation: Department of Hematological Medicine, Cell and Gene Therapy at King's, The Rayne Institute, King's College London, London, UK.
authors.forename: F
authors.initials: F
authors.lastname: Farzaneh
authors.affiliation: Department of Dermatology and Allergy, Skin Cancer Center Hannover, Hannover Medical School, Hannover, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Schenck
authors.affiliation: EUFETS GmbH, Idar-Oberstein, Heidelberg, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Naundorf
authors.affiliation: EUFETS GmbH, Idar-Oberstein, Heidelberg, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Kuehlcke
authors.affiliation: Division of Translational Oncology, National Center for Tumor Diseases, Heidelberg, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Ruggiero
authors.affiliation: Division of Translational Oncology, National Center for Tumor Diseases, Heidelberg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Division of Translational Oncology, National Center for Tumor Diseases, Heidelberg, Germany.
authors.forename: C
authors.initials: C
authors.lastname: von Kalle
authors.affiliation: Department of Experimental Hematology, Hannover, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Rothe
authors.affiliation: John Wayne Cancer Institute, Santa Monica, CA, USA.
authors.forename: D S B
authors.initials: DS
authors.lastname: Hoon
authors.affiliation: Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
authors.forename: L
authors.initials: L
authors.lastname: Gerasch
authors.affiliation: Department of Transfusion Medicine, Hannover Medical School, Hannover, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Figueiredo
authors.affiliation: Institute for Cell Therapeutics and GMP core facility IFB-Tx, Hannover Medical School, Hannover, Germany.
authors.forename: U
authors.initials: U
authors.lastname: Koehl
authors.affiliation: Department of Transfusion Medicine, Hannover Medical School, Hannover, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Blasczyk
authors.affiliation: Department of Dermatology and Allergy, Skin Cancer Center Hannover, Hannover Medical School, Hannover, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Gutzmer
authors.affiliation: Department of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, Hannover, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Stripecke
mesh_terms: D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D003713:Dendritic Cells / Q000276:immunology*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism*; D008545:Melanoma / Q000276:immunology / Q000628:therapy*; D008565:Membrane Proteins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology
chemical_list: D019496:Cancer Vaccines; D008565:Membrane Proteins; D010446:Peptide Fragments; C500290:peptide SVYDFFVWL