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Vectofusin-1 Improves Transduction of Primary Human Cells with Diverse Retroviral and Lentiviral Pseudotypes, Enabling Robust, Automated Closed-System Manufacturing

Authors
  • Radek, Constanze1
  • Bernadin, Ornellie2, 3, 4
  • Drechsel, Katharina1
  • Cordes, Nicole1, 5
  • Pfeifer, Rita1
  • Sträßer, Pia1
  • Mormin, Mirella6
  • Gutierrez-Guerrero, Alejandra2, 3, 4
  • Cosset, François-loïc2, 3, 4
  • Kaiser, Andrew D.1
  • Schaser, Thomas1
  • Galy, Anne6
  • Verhoeyen, Els2, 3, 4, 7
  • Johnston, Ian C.D.1
  • 1 Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany.
  • 2 CIRI—International Center for Infectiology Research, Team EVIR, Université de Lyon, Lyon, France.
  • 3 Inserm, U1111, Ecole Normale Supérieure de Lyon, Lyon, France.
  • 4 Université Lyon 1, CNRS, UMR5308, Lyon, France.
  • 5 Faculty of Biology, University of Freiburg, Freiburg, Germany.
  • 6 Integrare Research Unit UMR_S951, Genethon, INSERM, University Evry, EPHE, Evry, France.
  • 7 Université Côte d'Azur, INSERM, Centre Méditerranéen de Médecine Moléculaire (C3M), Nice, France.
Type
Published Article
Journal
Human Gene Therapy
Publisher
Mary Ann Liebert
Publication Date
Dec 01, 2019
Volume
30
Issue
12
Pages
1477–1493
Identifiers
DOI: 10.1089/hum.2019.157
PMID: 31578886
PMCID: PMC6919281
Source
PubMed Central
Keywords
License
Green

Abstract

Cell and gene therapies are finally becoming viable patient treatment options, with both T cell- and hematopoietic stem cell (HSC)-based therapies being approved to market in Europe. However, these therapies, which involve the use of viral vector to modify the target cells, are expensive and there is an urgent need to reduce manufacturing costs. One major cost factor is the viral vector production itself, therefore improving the gene modification efficiency could significantly reduce the amount of vector required per patient. This study describes the use of a transduction enhancing peptide, Vectofusin-1®, to improve the transduction efficiency of primary target cells using lentiviral and gammaretroviral vectors (LV and RV) pseudotyped with a variety of envelope proteins. Using Vectofusin-1 in combination with LV pseudotyped with viral glycoproteins derived from baboon endogenous retrovirus, feline endogenous virus (RD114), and measles virus (MV), a strongly improved transduction of HSCs, B cells and T cells, even when cultivated under low stimulation conditions, could be observed. The formation of Vectofusin-1 complexes with MV-LV retargeted to CD20 did not alter the selectivity in mixed cell culture populations, emphasizing the precision of this targeting technology. Functional, ErbB2-specific chimeric antigen receptor-expressing T cells could be generated using a gibbon ape leukemia virus (GALV)-pseudotyped RV. Using a variety of viral vectors and target cells, Vectofusin-1 performed in a comparable manner to the traditionally used surface-bound recombinant fibronectin. As Vectofusin-1 is a soluble peptide, it was possible to easily transfer the T cell transduction method to an automated closed manufacturing platform, where proof of concept studies demonstrated efficient genetic modification of T cells with GALV-RV and RD114-RV and the subsequent expansion of mainly central memory T cells to a clinically relevant dose.

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