Engineering Next-Generation BET-Independent MLV Vectors for Safer Gene Therapy

Overview

Journal Mol Ther Nucleic Acids

Publisher Cell Press

Date 2017 Jun 19

PMID 28624199

Citations 12

Authors

Sara El Ashkar

Dominique Van Looveren

Franziska Schenk

Lenard S Vranckx

Jonas Demeulemeester

Jan De Rijck

Zeger Debyser

Ute Modlich

Rik Gijsbers

Affiliations

Soon will be listed here.

Abstract

Retroviral vectors have shown their curative potential in clinical trials correcting monogenetic disorders. However, therapeutic benefits were compromised due to vector-induced dysregulation of cellular genes and leukemia development in a subset of patients. Bromodomain and extraterminal domain (BET) proteins act as cellular cofactors that tether the murine leukemia virus (MLV) pre-integration complex to host chromatin via interaction with the MLV integrase (IN) and thereby define the typical gammaretroviral integration distribution. We engineered next-generation BET-independent (Bin) MLV vectors to retarget their integration to regions where they are less likely to dysregulate nearby genes. We mutated MLV IN to uncouple BET protein interaction and fused it with chromatin-binding peptides. The addition of the CBX1 chromodomain to MLV IN efficiently targeted integration away from gene regulatory elements. The retargeted vector produced at high titers and efficiently transduced CD34 hematopoietic stem cells, while fewer colonies were detected in a serial colony-forming assay, a surrogate test for genotoxicity. Our findings underscore the potential of the engineered vectors to reduce the risk of insertional mutagenesis without compromising transduction efficiency. Ultimately, combined with other safety features in vector design, next-generation BinMLV vectors can improve the safety of gammaretroviral vectors for gene therapy.

Citing Articles

Long Terminal Repeats of Gammaretroviruses Retain Stable Expression after Integration Retargeting.

Miklik D, Slavkova M, Kucerova D, Mekadim C, Mrazek J, Hejnar J Viruses. 2024; 16(10).

PMID: 39459853 PMC: 11512309. DOI: 10.3390/v16101518.

Advances in HIV Gene Therapy.

Kitawi R, Ledger S, Kelleher A, Ahlenstiel C Int J Mol Sci. 2024; 25(5).

PMID: 38474018 PMC: 10931721. DOI: 10.3390/ijms25052771.

Determinants of Retroviral Integration and Implications for Gene Therapeutic MLV-Based Vectors and for a Cure for HIV-1 Infection.

Pellaers E, Bhat A, Christ F, Debyser Z Viruses. 2023; 15(1).

PMID: 36680071 PMC: 9861059. DOI: 10.3390/v15010032.

BET-Independent Murine Leukemia Virus Integration Is Retargeted and Selects Distinct Genomic Elements for Lymphomagenesis.

Nombela I, Michiels M, Van Looveren D, Marcelis L, El Ashkar S, Van Belle S Microbiol Spectr. 2022; 10(4):e0147822.

PMID: 35852337 PMC: 9431007. DOI: 10.1128/spectrum.01478-22.

Improved functionality and potency of next generation BinMLV viral vectors toward safer gene therapy.

Van Looveren D, Giacomazzi G, Thiry I, Sampaolesi M, Gijsbers R Mol Ther Methods Clin Dev. 2021; 23:51-67.

PMID: 34553002 PMC: 8433069. DOI: 10.1016/j.omtm.2021.07.003.

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