Efficient and robust NK-Cell transduction with Baboon Envelope pseudotyped lentivector: a major tool for immunotherapy

Ab, Colamartino; Lemieux, William; Bifsha, Panojot; Nicoletti, Simon; Chakravarti, Nitin; Js, Remon; Grisales-Romero, Hugo; Selleri, Silvia; Béland, Kathie; Guiot, Melanie; Tremblay‐Laganière, Camille; Dicaire, Renée; Barreiro, Luis B.; Da, Lee; Verhoeyen, Els; Haddad, Elias K.

doi:10.1101/625285

Cited by 6 publications

(6 citation statements)

References 20 publications

(21 reference statements)

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“…We found that activation of NK cells with IL-2 and IL-15 resulted in the upregulation of ASCT-2 expression, making them highly susceptible to transduction with BaEV pseudotyped LVs. Our findings are confirmed by a recent report by Colamartino et al (31).…”

Section: Discussionsupporting

confidence: 94%

A Distinct Subset of Highly Proliferative and Lentiviral Vector (LV)-Transducible NK Cells Define a Readily Engineered Subset for Adoptive Cellular Therapy

et al. 2019

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Genetic engineering is an important tool for redirecting the function of various types of immune cells and their use for therapeutic purpose. Although NK cells have many beneficial therapeutic features, genetic engineering of immune cells for targeted therapy focuses mostly on T cells. One of the major obstacles for NK cell immunotherapy is the lack of an efficient method for gene transfer. Lentiviral vectors have been proven to be a safe tool for genetic engineering, however lentiviral transduction is inefficient for NK cells. We show in this study that lentiviral vectors pseudotyped with a modified baboon envelope glycoprotein can transduce NK cells 20-fold or higher in comparison to VSV-G pseudotyped lentiviral vector. When we investigated the mechanism of transduction, we found that activated NK cells expressed baboon envelope receptor ASCT-2. Further analysis revealed that only a subset of NK cells could be expanded and transduced with an expression profile of NK56 bright , CD16 dim , TRAIL high , and CX3CR1 neg . Using CD19-CAR, we could show that CD19 redirected NK cells efficiently and specifically kill cell lines expressing CD19. Taken together, the results from this study will be important for future genetic modification and for redirecting of NK cell function for therapeutic purpose.

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Section: Discussionsupporting

confidence: 94%

A Distinct Subset of Highly Proliferative and Lentiviral Vector (LV)-Transducible NK Cells Define a Readily Engineered Subset for Adoptive Cellular Therapy

et al. 2019

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“…The authors would like to thank Dr. Dario Campana (St. Jude Children's Research Hospital) for the K562-mbIL15-41BBL cell line, and Dr. Rimas J. Orentas from the National Cancer Institute for the CAR-CD22 construct. This manuscript has been released as a Pre-Print at bioRxiv (20).…”

Section: Acknowledgmentsmentioning

confidence: 99%

Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector

et al. 2019

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NK-cell resistance to transduction is a major technical hurdle for developing NK-cell immunotherapy. By using Baboon envelope pseudotyped lentiviral vectors (BaEV-LVs) encoding eGFP, we obtained a transduction rate of 23.0 ± 6.6% (mean ± SD) in freshly-isolated human NK-cells (FI-NK) and 83.4 ± 10.1% (mean ± SD) in NK-cells obtained from the NK-cell Activation and Expansion System (NKAES), with a sustained transgene expression for at least 21 days. BaEV-LVs outperformed Vesicular Stomatitis Virus type-G (VSV-G)-, RD114-and Measles Virus (MV)-pseudotyped LVs (p < 0.0001). mRNA expression of both BaEV receptors, ASCT1 and ASCT2, was detected in FI-NK and NKAES, with higher expression in NKAES. Transduction with BaEV-LVs encoding for CAR-CD22 resulted in robust CAR-expression on 38.3 ± 23.8% (mean ± SD) of NKAES cells, leading to specific killing of NK-resistant pre-BALL -RS4;11 cell line. Using a larger vector encoding a dual CD19/CD22-CAR, we were able to transduce and re-expand dual-CAR-expressing NKAES, even with lower viral titer. These dual-CAR-NK efficiently killed both CD19 KO-and CD22 KO-RS4;11 cells. Our results suggest that BaEV-LVs may efficiently enable NK-cell biological studies and translation of NK-cell-based immunotherapy to the clinic.

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“…21 Culturing primary human NK cells on K562 feeder cells with expressing membrane-bound IL-21 and 4-1BBL also augmented gene transduction by 50%. 46,47 Third, suppression of intracellular antiviral defense mechanisms are also described to increase lentiviral transduction of NK cells. BX795, which is an inhibitor of the TBK1/IKKe complex that controls antiviral responses, is able to boost lentiviral gene transduction efficiency by 3.8-fold.…”

Section: Discussionmentioning

confidence: 99%

Rosuvastatin Enhances VSV-G Lentiviral Transduction of NK Cells via Upregulation of the Low-Density Lipoprotein Receptor

Gong

Wolterink

Janssen

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Adoptive natural killer (NK) cell therapy is attaining promising clinical outcomes in recent years, but improvements are needed. Genetic modification of NK cells with a tumor antigen-specific receptor on their surface coupled to intracellular signaling domains may lead to enhanced cytotoxicity against malignant cells. One of the most common approaches is by lentivirus-mediated transduction. However, NK cells are difficult to transduce and various methods have been attempted with different success rates. Because the low-density lipoprotein-receptor (LDLR) is the receptor of vesicular stomatitis virus (VSV) and is expressed only at low levels on NK cells, we tested the potential of 5 statins and 5 non-statin compounds to increase the LDLR expression, thereby facilitating viral transduction. We found that the transduction efficiency of VSV-G pseudotyped lentivirus is augmented by statins that induced higher LDLR expression. In both NK-92 cells and primary NK cells, the transduction efficiency increased after treatment with statins. Furthermore, statins have been reported to suppress NK cell cytotoxicity; however, we showed that this can be completely reversed by adding geranylgeranyl-pyrophosphate (GGPP). Among the statins tested, we found that the combination of rosuvastatin with GGPP most potently improved viral transduction without affecting the cytotoxic properties of the NK cells.

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Efficient and robust NK-Cell transduction with Baboon Envelope pseudotyped lentivector: a major tool for immunotherapy

Cited by 6 publications

References 20 publications

A Distinct Subset of Highly Proliferative and Lentiviral Vector (LV)-Transducible NK Cells Define a Readily Engineered Subset for Adoptive Cellular Therapy

A Distinct Subset of Highly Proliferative and Lentiviral Vector (LV)-Transducible NK Cells Define a Readily Engineered Subset for Adoptive Cellular Therapy

Efficient and Robust NK-Cell Transduction With Baboon Envelope Pseudotyped Lentivector

Rosuvastatin Enhances VSV-G Lentiviral Transduction of NK Cells via Upregulation of the Low-Density Lipoprotein Receptor

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