title: RD-MolPack technology for the constitutive production of self-inactivating lentiviral vectors pseudotyped with the nontoxic RD114-TR envelope.
abstract: To date, gene therapy with transiently derived lentivectors has been very successful to cure rare infant genetic diseases. However, transient manufacturing is unfeasible to treat adult malignancies because large vector lots are required. By contrast, stable manufacturing is the best option for high-incidence diseases since it reduces the production cost, which is the major current limitation to scale up the transient methods. We have previously developed the proprietary RD2-MolPack technology for the stable production of second-generation lentivectors, based on the RD114-TR envelope. Of note, opposite to vesicular stomatitis virus glycoprotein (VSV-G) envelope, RD114-TR does not need inducible expression thanks to lack of toxicity. Here, we present the construction of RD2- and RD3-MolPack cells for the production of self-inactivating lentivectors expressing green fluorescent protein (GFP) as a proof-of-concept of the feasibility and safety of this technology before its later therapeutic exploitation. We report that human T lymphocytes transduced with self-inactivating lentivectors derived from RD3-MolPack cells or with self-inactivating VSV-G pseudotyped lentivectors derived from transient transfection show identical T-cell memory differentiation phenotype and comparable transduction efficiency in all T-cell subsets. RD-MolPack technology represents, therefore, a straightforward tool to simplify and standardize lentivector manufacturing to engineer T-cells for frontline immunotherapy applications.
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Virna
authors.initials: V
authors.lastname: Marin
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Stornaiuolo
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Piovan
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Corna
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Bossi
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Monika
authors.initials: M
authors.lastname: Pema
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Giuliani
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Cinzia
authors.initials: C
authors.lastname: Scavullo
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Zucchelli
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Bordignon
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Gian Paolo
authors.initials: GP
authors.lastname: Rizzardi
authors.affiliation: Research Division, MolMed S.p.A , Milano, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bovolenta