Generation and Preclinical Characterization of a Fc-optimized GITR-Ig Fusion Protein for Induction of NK Cell Reactivity Against Leukemia

Schmiedel, Benjamin Joachim; Werner, Antje; Steinbacher, Julia; Nuebling, Tina; Buechele, Corina; Grosse‐Hovest, Ludger; Salih, Helmut R.

doi:10.1038/mt.2013.11

Cited by 17 publications

(20 citation statements)

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“…DAC treatment additionally increases expression of Tnfsf18 (GitrL), which is the ligand for the T-cell costimulatory molecule GITR. GITR ligation enhances the infiltration and activity of effector T cells (33) and NK cells (22,34) and generates effector T cells that are resistant to the immunosuppressive effects of Tregs (35). These data suggest that DAC not only has cancer cellintrinsic antitumor effects but also modifies the tumor microenvironment to promote T-cell infiltration and antitumor function.…”

Section: Discussionmentioning

confidence: 88%

Decitabine Enhances Lymphocyte Migration and Function and Synergizes with CTLA-4 Blockade in a Murine Ovarian Cancer Model

Wang

Amoozgar

Huang

et al. 2015

Cancer Immunology Research

143

105

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The lack of second-line treatment for relapsed ovarian cancer necessitates the development of improved combination therapies. Targeted therapy and immunotherapy each confer clinical benefit, albeit limited as monotherapies. Ovarian cancer is not particularly responsive to immune checkpoint blockade, so combination with a complementary therapy may be beneficial. Recent studies have revealed that a DNA methyl transferase inhibitor, azacytidine, alters expression of immunoregulatory genes in ovarian cancer. In this study, the antitumor effects of a related DNA methyl transferase inhibitor, decitabine (DAC), were demonstrated in a syngeneic murine ovarian cancer model. Low-dose DAC treatment increases the expression of chemokines that recruit NK cells and CD8 þ T cells, promotes their production of IFNg and TNFa, and extends the survival of mice bearing subcutaneous or orthotopic tumors. While neither DAC nor immune checkpoint blockade confers durable responses as a monotherapy in this model, the efficacy of anti-CTLA-4 was potentiated by combination with DAC. This combination promotes differentiation of na€ ve T cells into effector T cells and prolongs cytotoxic lymphocyte responses as well as mouse survival. These results suggest that this combination therapy may be worthy of further consideration for improved treatment of drug-resistant ovarian cancer.

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Section: Discussionmentioning

confidence: 88%

Decitabine Enhances Lymphocyte Migration and Function and Synergizes with CTLA-4 Blockade in a Murine Ovarian Cancer Model

Wang

Amoozgar

Huang

et al. 2015

Cancer Immunology Research

143

105

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“…Antibodies against tumor antigens (e.g., CD20 and CS1) can induce ADCC of NK cells ( 74 , 75 ). Antibodies to activating NK receptors (e.g., 4-1BB, GITR, NKG2D, DNAM-1, and NCRs) can enhance NK activation ( 74 , 76 – 79 ). In addition, inhibitory receptors (e.g., KIR2DL, PD-1, PD-L1, and NKG2A) can be blocked by antibodies ( 80 – 85 ).…”

Section: Future Directionsmentioning

confidence: 99%

Present and Future of Allogeneic Natural Killer Cell Therapy

Lim¹,

Jung²,

Hwang

et al. 2015

Front. Immunol.

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Natural killer (NK) cells are innate lymphocytes that are capable of eliminating tumor cells and are therefore used for cancer therapy. Although many early investigators used autologous NK cells, including lymphokine-activated killer cells, the clinical efficacies were not satisfactory. Meanwhile, human leukocyte antigen (HLA)-haploidentical hematopoietic stem cell transplantation revealed the antitumor effect of allogeneic NK cells, and HLA-haploidentical, killer cell immunoglobulin-like receptor ligand-mismatched allogeneic NK cells are currently used for many protocols requiring NK cells. Moreover, allogeneic NK cells from non-HLA-related healthy donors have been recently used in cancer therapy. The use of allogeneic NK cells from non-HLA-related healthy donors allows the selection of donor NK cells with higher flexibility and to prepare expanded, cryopreserved NK cells for instant administration without delay for ex vivo expansion. In cancer therapy with allogeneic NK cells, optimal matching of donors and recipients is important to maximize the efficacy of the therapy. In this review, we summarize the present state of allogeneic NK cell therapy and its future directions.

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“…NKG2D–Fc fusion proteins were generated as dimeric constructs as described previously . SP2/0‐Ag14 cells (ATCC, Manassas, VA) were transfected with vectors coding for the different NKG2D–Fc constructs or Fc controls by electroporation.…”

Section: Methodsmentioning

confidence: 99%

“…NKG2D-Fc fusion proteins were generated as dimeric constructs as described previously. 26,27 SP2/0-Ag14 cells (ATCC, Manassas, VA) were transfected with vectors coding for the different NKG2D-Fc constructs or Fc controls by electroporation. Constructs were purified from culture supernatants by Protein A affinity chromatography and purity was determined by SDS-PAGE and size exclusion chromatography using a Superdex 200 PC3.2/30 column (SMART System, GE Healthcare, Dornstadt, Germany).…”

Section: Production and Purification Of Nkg2d-fc Fusion Proteins Andmentioning

confidence: 99%

An Fc‐optimized NKG2D‐immunoglobulin G fusion protein for induction of natural killer cell reactivity against leukemia

Steinbacher

Baltz-Ghahremanpour

Schmiedel

et al. 2014

Intl Journal of Cancer

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Recruitment of Fc-receptor-bearing effector cells, such as natural killer (NK) cells, is a feature critical for the therapeutic success of antitumor antibodies and can be improved by the modifications of an antibody's Fc part. The various ligands of the activating immunoreceptor NKG2D, NKG2DL) are selectively expressed on malignant cells including leukemia. We here took advantage of the tumor-associated expression of NKG2DL for targeting leukemic cells by NKG2D-immunoglobulin G (IgG)1 fusion proteins containing modified Fc parts. Compared to NKG2D-Fc containing a wild-type Fc part (NKG2D-Fc-WT), our mutants (S239D/I332E and E233P/L234V/L235A/DG236/A327G/A330S) displayed highly enhanced (NKG2D-Fc-ADCC) and abrogated (NKG2D-Fc-KO) affinity to the NK cell Fc receptor, respectively. Functional analyses with allogenic as well as autologous NK cells and primary malignant cells of leukemia patients revealed that NKG2D-Fc-KO significantly reduced NK reactivity by blocking immunostimulatory NKG2D-NKG2DL interaction. NKG2D-Fc-WT already enhanced antileukemia reactivity by inducing antibody-dependent cellular cytotoxicity (ADCC) with NKG2D-Fc-ADCC mediating significantly stronger effects. Parallel application of NKG2D-Fc-ADCC with Rituximab caused additive effects in lymphoid leukemia. In line with the tumorassociated expression of NKG2DL, no NK cell ADCC against resting healthy blood cells was induced. Thus, NKG2D-Fc-ADCC potently enhances NK antileukemia reactivity despite the inevitable reduction of activating signals upon binding to NKG2DL and may constitute an attractive means for immunotherapy of leukemia.For more than a decade, chimeric or humanized secondgeneration monoclonal antibodies have been successfully used in cancer therapy. Introduction, for example, of Rituximab and Herceptin in therapeutic regimes applied to patients with B-cell Non-Hodgkin's lymphoma and Her2-positive breast cancer, respectively, lead to considerably improved outcomes. 1,2 Nevertheless, the "antibody success story" has its shortcomings: some patients do not respond at all, others for a limited time only. Thus, numerous strategies are presently being evaluated to increase the efficacy of antitumor antibodies. 3,4 To this end, improving the ability to recruit Fcreceptor (FcR)-bearing immune cells, one of the most important antibody functions constitutes a highly promising approach. 5,6 Currently, three antibodies, directed to CD19, CD30 and CD40, carrying the amino acid modifications S239D and I332E (SDIE modification) 7-9 and a CD20 antibody with improved, low-fucose glycosylation (GA101) 10 are in early clinical development. These reagents mediate markedly enhanced antibody-dependent cellular cytotoxicity (ADCC) against tumor cells, and it appears that their

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Generation and Preclinical Characterization of a Fc-optimized GITR-Ig Fusion Protein for Induction of NK Cell Reactivity Against Leukemia

Cited by 17 publications

References 50 publications

Decitabine Enhances Lymphocyte Migration and Function and Synergizes with CTLA-4 Blockade in a Murine Ovarian Cancer Model

Decitabine Enhances Lymphocyte Migration and Function and Synergizes with CTLA-4 Blockade in a Murine Ovarian Cancer Model

Present and Future of Allogeneic Natural Killer Cell Therapy

An Fc‐optimized NKG2D‐immunoglobulin G fusion protein for induction of natural killer cell reactivity against leukemia

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