Comparison of mRNA and lentiviral based transfection of natural killer cells with chimeric antigen receptors recognizing lymphoid antigens

Boissel, Laurent; Betancur, Monica; Lu, Weiquan; Wels, Winfried S.; Marino, Tiziana; Etten, Richard A. Van; Klingemann, Hans

doi:10.3109/10428194.2011.634048

Cited by 110 publications

(103 citation statements)

References 37 publications

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“…Lentiviral vectors have moderate levels of insertion mutations, but their lentiviral transduction efficiency is very low for PBderived NK cells (between 8%-16%). However, the transduction efficiency of cord blood-derived NK cells reaches 73% [65] , which is sufficient to introduce CARs into cord blood-derived NK cells. Nevertheless, the lentiviral transduction efficiency of PB-derived NK cells requires improvement.…”

Section: Methods For Car Transduction Into Nk Cellsmentioning

confidence: 99%

Chimeric antigen receptor (CAR)-transduced natural killer cells in tumor immunotherapy

2017

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Natural killer (NK) cells are potential effector cells in cell-based cancer immunotherapy, particularly in the control of hematological malignancies. The chimeric antigen receptor (CAR) is an artificially modified fusion protein that consists of an extracellular antigen recognition domain fused to an intracellular signaling domain. T cells genetically modified with a CAR have demonstrated remarkable success in the treatment of hematological cancers. Compared to T cells, CAR-transduced NK cells (CAR-NK) exhibit several advantages, such as safety in clinical use, the mechanisms by which they recognize cancer cells, and their abundance in clinical samples. Human primary NK cells and the NK-92 cell line have been successfully transduced to express CARs against both hematological cancers and solid tumors in pre-clinical and clinical trials. However, many challenges and obstacles remain, such as the ex vivo expansion of CAR-modified primary NK cells and the low transduction efficiency of NK cells. Many strategies and technologies have been developed to improve the safety and therapeutic efficacy in CAR-based immunotherapy. Moreover, NK cells express a variety of activating receptors (NKRs), such as CD16, NKG2D, CD226 and NKp30, which might specifically recognize the ligands expressed on tumor cells. Based on the principle of NKR recognition, a strategy that targets NKRs is rapidly emerging. Given the promising clinical progress described in this review, CAR- and NKR-NK cell-based immunotherapy are likely promising new strategies for cancer therapy.

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Section: Methods For Car Transduction Into Nk Cellsmentioning

confidence: 99%

Chimeric antigen receptor (CAR)-transduced natural killer cells in tumor immunotherapy

2017

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“…As an alternative to T cells, natural killer (NK) cells expressing CARs directed against CD19 or CD20 (2 markers of the B-cell lineage) can also be used as cytotoxic effector cells for cell-based immunotherapy. However, current NK cell-based therapies are constrained by the necessity to isolate sufficient numbers of NK cells from donors as well as by the need to achieve acceptable transfection efficiencies 6 , 7 . The human NK cell line NK-92 8 , 9 presents an attractive alternative to donor NK cells, as it can be propagated and expanded in vitro 10 .…”

Section: Introductionmentioning

confidence: 99%

Retargeting NK-92 cells by means of CD19- and CD20-specific chimeric antigen receptors compares favorably with antibody-dependent cellular cytotoxicity

et al. 2013

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Multiple natural killer (NK) cell-based anticancer therapies are currently under development. Here, we compare the efficiency of genetically modified NK-92 cells expressing chimeric antigen receptors (CARs) at killing NK cell-resistant B-lymphoid leukemia cells to the antibody-dependent cell-mediated cytotoxicity (ADCC) of NK-92 cells expressing a high affinity variant of the IgG Fc receptor (FcγRIII). First, we compared in vitro the abilities of NK-92 cells expressing CD20-targeting CARs to kill primary chronic lymphocytic leukemia (CLL) cells derived from 9 patients with active, untreated disease to the cytotoxicity of NK-92 cells expressing FcγRIII combined with either of the anti-CD20 monoclonal antibodies (mAbs) rituximab or ofatumumab. We found that CAR-expressing NK-92 cells effectively kill NK cell-resistant primary CLL cells and that such a cytotoxic response is significantly stronger than that resulting from ADCC. For studying CAR-expressing NK cell-based immunotherapy in vivo, we established xenograft mouse models of residual leukemia using the human BCR-ABL1+ cell lines SUP-B15 (CD19+CD20−) and TMD-5 (CD19+CD20+), two acute lymphoblastic leukemia (ALL) lines that are resistant to parental NK-92 cells. Intravenous injection of NK-92 cells expressing CD19-targeting CARs eliminated SUP-B15 cells, whereas they had no such effect on TMD-5 cells. However, the intrafemoral injection of NK-92 cells expressing CD19-targeting CAR resulted in the depletion of TMD-5 cells from the bone marrow environment. Comparative studies in which NK-92 cells expressing either CD19- or CD20-targeting CARs were directly injected into subcutaneous CD19+CD20+ Daudi lymphoma xenografts revealed that CD20-targeting CAR is superior to its CD19-specific counterpart in controlling local tumor growth. In summary, we show here that CAR-expressing NK-92 cells can be functionally superior to ADCC (as mediated by anti-CD20 mAbs) in the elimination of primary CLL cells. Moreover, we provide data demonstrating that the systemic administration of CAR-expressing NK-92 cells can control lymphoblastic leukemia in immunocompromised mice. Our results also suggest that the direct injection of CAR-expressing NK-92 cells to neoplastic lesions could be an effective treatment modality against lymphoma.

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“…Although LVs have been used to efficiently transfer genes into human T cells and the NK cell lines such as NK92 and NKL, LV transduction of fresh and ex vivo-expanded human NK cells has been more challenging. 73 We used an LV-expressing enhanced green fluorescence protein driven by a murine stem cell virus long terminal repeat promoter to transduce CD3 Ϫ and CD56 ϩ and/or CD16 ϩ human NK cells that were either resting, IL-2 activated, or expanded ex vivo using an irradiated EBV-LCL feeder cells. 4 Resting NK cells were difficult to transduce with LVs, even at high multiplicities of infection, with transduction efficiencies in the range of only 3% to 14%.…”

Section: Future Directions: Engineering a Better Nk Cellmentioning

confidence: 99%

“…These data, as well as recent data showing the efficacy of T-cell-based CAR therapy targeting CD19 in B-cell malignancies, suggest that NK cells modified to express CARs after mRNA or viral transduction represent a therapeutic tool worthy of exploration in the clinical setting. 43,44,57,73,[76][77][78][79] The ability of CAR-expressing NK cells to home to the tumor may be a critical determinant of their efficacy in humans. As discussed previously, expanded NK cells may lack or down-regulate molecules that are critical for NK cell homing from the circulation.…”

Section: Future Directions: Engineering a Better Nk Cellmentioning

confidence: 99%

Bringing natural killer cells to the clinic: ex vivo manipulation

Childs

Berg

2013

Hematology

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Recently, there has been a substantial gain in our understanding of the role that natural killer (NK) cells play in mediating innate host immune responses against viruses and cancer. Although NK cells have long been known to be capable of killing cancer cells independently of antigen recognition, the full therapeutic potential of NK cell-based immunotherapy has yet to be realized. Here we review novel methods to activate and expand human NK cells ex vivo for adoptive transfer in humans, focusing on the important phenotypic and functional differences observed among freshly isolated, cytokine activated, and ex vivo-expanded NK populations. Natural killer cell therapy for cancer: a new hopeThe ability of natural killer (NK) cells to kill tumor cells without the need to recognize a tumor-specific antigen provides advantages over T cells and makes them appealing for investigation as effectors for immunotherapy. 1,2 There has recently been a substantial gain in our understanding of the role that NK cells play in mediating innate host immune responses against viruses and cancer. Although NK cells have long been known to be capable of killing cancer cells independently of antigen recognition, the full therapeutic potential of NK cell-based immunotherapy has yet to be realized. Here we review novel methods to activate and expand human NK cells ex vivo for adoptive transfer in humans, focusing on the important phenotypic and functional differences observed among freshly isolated, cytokine activated, and ex vivo-expanded NK populations. Ex vivo NK cell activation and expansionUnlike T cells, NK cells represent only a minor fraction of human lymphocytes. NK cells are defined by their CD3 Ϫ /CD56 ϩ phenotype, comprising 5% to 15% of circulating lymphocytes, and are commonly divided into CD56 dim CD16 ϩ (90%) and CD56 bright CD16 Ϫ (10%) subpopulations with distinct effector functions. Recently, investigators have shown that NK cell tumor cytotoxicity can be enhanced by disrupting NK cell signaling through inhibitory receptors such as KIR, PD-1, and NKG2A. 3,4 Furthermore, exposing tumors to drugs that down-regulate ligands for NK cell inhibitory receptors or up-regulate death receptors for NK cell effector molecules or ligands that bind NK cell-activating receptors can be used as a strategy to sensitize tumors to NK cell-mediated apoptosis. Recently, anti-PD-1 and anti-PD-L1 monoclonal antibodies and the immunomodulatory drug lenalidomide were shown to enhance both NK cell tumor trafficking and NK cell-mediated antibody-dependent cell-mediated cytotoxicity, and cytokine release against tumors while simultaneously suppressing regulatory T cell function. [5][6][7][8] Similarly, anti-CTLA-4 monoclonal antibodies have been shown to augment NK cell antibody-dependent cellmediated cytotoxicity and TNF␣ release against melanoma cells by Fc␥RIII (CD16) binding to antibody-bound tumor cells, as well as through regulatory T cell inactivation. 9-11 These data suggest these agents could be used in conjunction with autologous NK cell inf...

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Comparison of mRNA and lentiviral based transfection of natural killer cells with chimeric antigen receptors recognizing lymphoid antigens

Cited by 110 publications

References 37 publications

Chimeric antigen receptor (CAR)-transduced natural killer cells in tumor immunotherapy

Chimeric antigen receptor (CAR)-transduced natural killer cells in tumor immunotherapy

Retargeting NK-92 cells by means of CD19- and CD20-specific chimeric antigen receptors compares favorably with antibody-dependent cellular cytotoxicity

Bringing natural killer cells to the clinic: ex vivo manipulation

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