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Preclinical Evaluation of a Novel TALEN Targeting CCR5 Confirms Efficacy and Safety in Conferring Resistance to HIV‐1 Infection
Author(s) -
Romito Marianna,
Juillerat Alexandre,
Kok Yik Lim,
Hildenbeutel Markus,
Rhiel Manuel,
Andrieux Geoffroy,
Geiger Johannes,
Rudolph Carsten,
Mussolino Claudio,
Duclert Aymeric,
Metzner Karin J.,
Duchateau Philippe,
Cathomen Toni,
Cornu Tatjana I.
Publication year - 2021
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.202000023
Subject(s) - transcription activator like effector nuclease , genome editing , biology , effector , primary immunodeficiency , cxcr4 , virology , transplantation , chemokine receptor , stem cell , crispr , immunology , gene , genetics , chemokine , receptor , medicine , immune system , surgery
Therapies to treat patients infected with human immunodeficiency virus (HIV) aim at preventing viral replication but fail to eliminate the virus. Although transplantation of allogeneic CCR5 Δ32 homozygous stem cell grafts provided a cure for a few patients, this approach is not considered a general therapeutic strategy because of potential side effects. Conversely, gene editing to disrupt the C‐C chemokine receptor type 5 (CCR5) locus, which encodes the major HIV coreceptor, has shown to confer resistance to CCR5‐tropic HIV strains. Here, an engineered transcription activator‐like effector nuclease (TALEN) that enables efficient CCR5 editing in hematopoietic cells is presented. After transferring TALEN‐encoding mRNA into primary CD4+ T cells, up to 89% of CCR5 alleles are disrupted. Genotyping confirms the genetic stability of the CCR5 ‐edited cells, and genome‐wide off‐target analyses established the absence of relevant mutagenic events. When challenging the edited T cells with CCR5‐tropic HIV, protection in a dose‐dependent manner is observed. Functional assessments reveal no significant differences between edited and control cells in terms of proliferation and their ability to secrete cytokines upon exogenous stimuli. In conclusion, a highly active and specific TALEN to disrupt CCR5 is successfully engineered, paving the way for its clinical application in hematopoietic stem cell grafts.