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Efficient and Highly Specific Gene Transfer Using Mutated Lentiviral Vectors Redirected with Bispecific Antibodies
- Source :
- mBio, Vol 11, Iss 1 (2020), mBio, Vol 11, Iss 1, p e02990-19 (2020), mBio
- Publication Year :
- 2020
- Publisher :
- American Society for Microbiology, 2020.
-
Abstract
- The goal of gene therapy is specific delivery and expression of therapeutic genes to target cells and tissues. Common lentivirus (LV) vectors are efficient gene delivery vehicles but offer little specificity. Here, we report an effective and versatile strategy to redirect LV to target cells using bispecific antibodies (bsAbs) that bind both cell receptors and LV envelope domains. Importantly, we ablated the native receptor binding of LV to minimize off-target transduction. Coupling bsAb specificity and ablated native LV tropism synergistically enhanced the selectivity of our targeted gene delivery system. The modular nature of our bsAb-based redirection enables facile targeting of the same LV to diverse tissues/cells. By abrogating the native broad tropism of LV, our bsAb-LV redirection strategy may enable lentivirus-based gene delivery in vivo, expanding the current use of LV beyond ex vivo applications.<br />Despite their exceptional potencies, the broad tropism of most commonly used lentivirus (LV) vectors limits their use for targeted gene delivery in vivo. We hypothesized that we could improve the specificity of LV targeting by coupling (i) reduction of their binding to off-target cells with (ii) redirection of the vectors with a bispecific antibody (bsAb) that binds both LV and receptors on target cells. As a proof of concept, we pseudotyped nonreplicating LV using a mutated Sindbis envelope (mSindbis) with ablated binding to native receptors, while retaining the capacity to facilitate efficient fusion and endosomal escape. We then evaluated the transduction potencies of the mSindbis LV for HER2-positive (HER2+) (SKBR3) breast and HER2-negative (HER2−) (A2780) cells when redirected with different bsAbs. mSindbis LV alone failed to induce appreciable green fluorescent protein (GFP) expression in either cell. When mixed with HER2-targeting bsAb, mSindbis LV was exceptionally potent, transducing 12% to 16% of the SKBR3 cells at a multiplicity of infection (MOI [ratio of viral genome copies to target cells]) of 3. Transduction was highly specific, resulting in ∼50-fold-greater selectivity toward SKBR3 cells versus A2780 cells. Redirecting mSindbis LV led to a 10-fold improvement in cell-specific targeting compared to redirecting wild-type Sindbis LV with the same bsAb, underscoring the importance of ablating native virus tropism in order to maximize targeting specificity. The redirection of mutated LV using bsAb represents a potent and highly versatile platform for targeted gene therapy.
- Subjects :
- Molecular Biology and Physiology
Genetic enhancement
Genetic Vectors
Gene delivery
targeted gene delivery
Microbiology
Green fluorescent protein
03 medical and health sciences
Transduction (genetics)
0302 clinical medicine
Multiplicity of infection
Antibody Specificity
Transduction, Genetic
lentivirus
Cell Line, Tumor
Virology
Antibodies, Bispecific
Sindbis
Biomarkers, Tumor
Humans
Antigens
Tropism
030304 developmental biology
0303 health sciences
biology
Chemistry
Gene Transfer Techniques
Genetic Therapy
biology.organism_classification
gene therapy
QR1-502
Cell biology
bispecific antibody
030220 oncology & carcinogenesis
Mutation
Lentivirus
Ex vivo
Research Article
Protein Binding
Subjects
Details
- Language :
- English
- ISSN :
- 21507511
- Volume :
- 11
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- mBio
- Accession number :
- edsair.doi.dedup.....481da69605bbc77a928cb73237daf2c3