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Clinically relevant CCR5 editing in primary CD4+ T cells with TALEN confers resistance to HIV-1 infection
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  • Marianna Romito,
  • Alexandre Juillerat,
  • Yik Lim Kok,
  • Markus HIldenbeutel,
  • Manuel Rhiel,
  • Geoffroy Andrieux,
  • Johannes Geiger,
  • Carsten Rudolph,
  • Claudio Mussolino,
  • Aymeric Duclert,
  • Karin Metzner,
  • Philippe Duchateau,
  • Toni Cathomen,
  • Tatjana Cornu
Marianna Romito
University Medical Center Freiburg
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Alexandre Juillerat
Cellectis Inc.
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Yik Lim Kok
University Hospital Zurich Division of Infectious Diseases and Hospital Epidemiology
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Markus HIldenbeutel
University Medical Center Freiburg
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Manuel Rhiel
University Medical Center Freiburg
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Geoffroy Andrieux
University Medical Center Freiburg
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Johannes Geiger
ethris GmbH
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Carsten Rudolph
ethris GmbH
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Claudio Mussolino
University Medical Center Freiburg
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Aymeric Duclert
Cellectis SA
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Karin Metzner
University Hospital Zurich Division of Infectious Diseases and Hospital Epidemiology
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Philippe Duchateau
Cellectis SA
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Toni Cathomen
University Medical Center Freiburg
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Tatjana Cornu
University Medical Center Freiburg
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Abstract

Today’s available therapies to treat patients infected with human immunodeficiency virus (HIV) aim at preventing viral replication and transmission but fail to eliminate the virus. Although transplantation of an allogeneic CCR5Δ32 homozygous stem cell grafts provided a cure for three patients, this approach is not considered a general therapeutic strategy because of potential severe side effects. Conversely, genome editing to disrupt the CCR5 locus that encodes the major HIV coreceptor was shown to confer resistance to R5-tropic HIV strains on the cellular level. Here, we present a clinically relevant and highly efficient approach to produce HIV-1 resistant CD4+ T cells. After transferring mRNA coding for CCR5-targeting TALEN into CD4+ T cells by electroporation, up to 89% of CCR5 alleles were disrupted. Genotyping confirmed genetic stability of the edited cell product and off-target analyses established absence of relevant mutagenic events. When challenging these edited T cells with R5-tropic HIV, we observed protection in a dose-dependent manner. Functional assessments revealed no significant differences between edited and control CD4+ T cells in terms of proliferation capacity and their ability to secrete cytokines upon exogenous stimuli. Overall, we successfully engineered HIV-resistant CD4+ T cells under clinically relevant conditions, paving the way for clinical translation.

Peer review status:UNDER REVIEW

18 May 2020Submitted to Biotechnology Journal
19 May 2020Submission Checks Completed
19 May 2020Assigned to Editor
19 May 2020Reviewer(s) Assigned
20 Aug 2020Editorial Decision: Revise Major
23 Sep 20201st Revision Received
23 Sep 2020Submission Checks Completed
23 Sep 2020Assigned to Editor
23 Sep 2020Reviewer(s) Assigned