Abstract Triple-negative breast cancer (TNBC) is an aggressive disease lacking targeted therapy. In this study, we developed a CAR T cell–based immunotherapeutic strategy to target TEM8, a marker initially defined on endothelial cells in colon tumors that was discovered recently to be upregulated in TNBC. CAR T cells were developed that upon specific recognition of TEM8 secreted immunostimulatory cytokines and killed tumor endothelial cells as well as TEM8-positive TNBC cells. Notably, the TEM8 CAR T cells targeted breast cancer stem–like cells, offsetting the formation of mammospheres relative to nontransduced T cells. Adoptive transfer of TEM8 CAR T cells induced regression of established, localized patient-derived xenograft tumors, as well as lung metastatic TNBC cell line–derived xenograft tumors, by both killing TEM8+ TNBC tumor cells and targeting the tumor endothelium to block tumor neovascularization. Our findings offer a preclinical proof of concept for immunotherapeutic targeting of TEM8 as a strategy to treat TNBC. Significance: These findings offer a preclinical proof of concept for immunotherapeutic targeting of an endothelial antigen that is overexpressed in triple-negative breast cancer and the associated tumor vasculature. Cancer Res; 78(2); 489–500. ©2017 AACR.

Triple-negative breast cancer

Triple negative

10.1158/0008-5472.can-16-1911

Cite

Citations (136)

40P Connexin43 as double therapeutic target in triple-negative breast cancer: Role in tumour progression and antitumour immune response

Annals of Oncology (2022)

Marina Rodríguez-Candela Mateos Jasmin Röder Pranav Oberoi Winfried S. Wels Paz Santiago-Freijanes

Triple-negative breast cancer

10.1016/j.annonc.2022.03.055

Cite

Citations (0)

Continuously expanding CAR NK-92 cells display selective cytotoxicity against B-cell leukemia and lymphoma

Cytotherapy (2016)

Sarah Oelsner Miriam E. Friede Congcong Zhang Juliane Wagner Susanne Badura

NK-92

Janus kinase 3

Lymphokine-activated killer cell

CD137

10.1016/j.jcyt.2016.10.009

Cite

Citations (169)

Expression of an oncogenic mutant EGF receptor markedly increases the sensitivity of cells to an EGF‐receptor‐specific antibody‐toxin

International Journal of Cancer (1998)

Mathias Schmidt Pia Reiser David A. Hills William J. Gullick Winfried S. Wels

EGFRvIII is a ligand-independent, constitutively active variant of the epidermal growth factor receptor (EGFR) that is specifically expressed in gliomas and various other human malignancies and has been proposed as a target for directed tumor therapy. We have recently constructed a highly potent single-chain antibody-toxin, scFv(14E1)-ETA, which consists of the variable domains of the antibody 14E1 specific for human full-length EGFR genetically fused to a truncated form of Pseudomonas exotoxin A. We demonstrate here binding of 14E1 antibody to both full-length and variant EGFR. In contrast to a recombinant toxin containing transforming growth factor-alpha (TGF-alpha) as a cell targeting domain, scFv(14E1)-ETA was highly active on cells expressing EGFRvIII. Surprisingly, scFv(14E1)-ETA displayed cell killing activity on EGFRvIII-expressing cells that was up to 100-fold higher than on control cells expressing full-length EGFR. No differences in the binding affinities of scFv(14E1)-ETA to full-length EGFR or EGFRvIII were observed, suggesting that events downstream of immunotoxin binding are responsible for the increased sensitivity of EGFRvIII-expressing cells. This might have implications for the development of therapeutic reagents simultaneously targeting different forms of the EGFR.

Pseudomonas exotoxin

Diphtheria Toxin

10.1002/(sici)1097-0215(19980316)75:6<878::aid-ijc10>3.0.co;2-#

Cite

Citations (30)

Cancer Cell International (2004)

Florian Rohrbach Robert Weth Mischo Kursar H-W Mittrücker Winfried S. Wels

Tumor-specific T lymphocytes can be regarded as a highly effective mechanism for tumor rejection. A substantial number of T-cell defined tumor antigens including mutated oncoproteins and differentiation antigens have been identified. However, while most spontaneous tumors appear to be antigenic, few are immunogenic. Activation of tumor-specific cytotoxic T cells (CTL) requires presentation of tumor antigens by professional antigen presenting cells (APCs) via MHC I molecules. Due to their crucial role in T-cell activation, APCs are being exploited for active cancer immunotherapy. Present experimental strategies include the incubation of dendritic cells with synthetic, tumor specific peptides to achieve uptake of tumor antigens and presentation in the context of MHC molecules. Alternatively, gene therapeutic approaches are aimed at the endogenous expression of tumor antigens in APCs upon transfer of suitable vector constructs.

10.1186/1475-2867-4-s1-s36

Cite

Citations (0)

Surface Charge-Modification Prevents Sequestration and Enhances Tumor-Cell Specificity of a Recombinant Granzyme B–TGFα Fusion Protein

Bioconjugate Chemistry (2012)

Robert A. Jabulowsky Pranav Oberoi Hayat Bähr-Mahmud Benjamin Dälken Winfried S. Wels

The serine protease granzyme B (GrB) plays an important role in the immune defense mediated by cytotoxic lymphocytes. Recombinant derivatives of this pro-apoptotic protein fused to tumor-targeting ligands hold promise for cancer therapy, but their applicability may be limited by promiscuous binding to nontarget tissues via electrostatic interactions. Here, we investigated cell binding and specific cytotoxicity of chimeric molecules consisting of wild-type or surface-charge-modified human GrB and the natural EGFR ligand TGFα for tumor targeting. We mutated two cationic heparin-binding motifs responsible for electrostatic interactions of GrB with cell surface structures, and genetically fused the resulting GrBcs derivative to TGFα for expression in the yeast Pichia pastoris. Purified GrBcs-TGFα (GrBcs-T) and a corresponding fusion protein employing wild-type GrB (GrB-T) displayed similar enzymatic activity and targeted cytotoxicity against EGFR-overexpressing breast carcinoma cells in the presence of an endosomolytic reagent. However, unspecific binding of the modified GrBcs-T variant to EGFR-negative cells was dramatically reduced, preventing the sequestration by nontarget cells in mixed cell cultures and increasing tumor-cell specificity. Likewise, modification of the GrB domain alleviated unspecific extracellular effects such as cell detachment indicative of extracellular matrix degradation. Our data demonstrate improved selectivity and functionality of surface-charge-modified GrBcs, suggesting this strategy as a general approach for the development of optimized GrB fusion proteins for therapeutic applications.

Granzyme

10.1021/bc3000657

Cite

Citations (13)

EGFR/EGFRvIII-targeted CAR-NK cells as a promising allogeneic cell therapy for the treatment of solid tumors

Klinische Pädiatrie (2024)

Leonhard Reindl S Thul Barbra A. Roller Halvard Bönig Jan‐Henning Klusmann

Targeted Therapy

Cell therapy

10.1055/s-0044-1786612

Cite

Citations (0)

566. Selective and Stable Transduction of Human CD4+ T Cells In Vivo Upon Systemic Administration of CD4-Targeted Lentiviral Vectors

Molecular Therapy (2015)

Qi Zhou Katharina M. Uhlig Anke Muth Janine Kimpel Camille Lévy

Playing a central role in both innate and adaptive immunity, CD4+ T cells are the key target for genetic modifications in basic research and immunotherapy. Specific and stable delivery of therapeutic genes into these cells is therefore highly desirable. Here, we describe novel lentiviral vectors (CD4-LV) that have been rendered selective for human or simian CD4+ cells by surface engineering. This novel CD4-LV was highly specific and effective in genetic modification of human CD4+ T cells both in vitro and in vivo. When applied to peripheral blood mononuclear cells (PBMC), CD4-LV transduced CD4+ but not CD4− cells. Notably, also unstimulated T cells were stably genetically modified. Upon systemic or intrasplenic administration into mice reconstituted with human PBMC or hematopoietic stem cells, reporter gene expression was predominantly detected in lymphoid organs. Evaluation of GFP expression in organ-derived cells and blood by flow cytometry demonstrated exclusive gene transfer into CD4+ human lymphocytes. In bone marrow and spleen memory T cells were preferentially hit. Towards potential therapeutic applications, we show that CD4-LV can deliver genes protecting T cells from infection with HIV-1 or encoding chimeric antigen receptors thereby inducing tumor cell killing. This study demonstrates for the first time highly selective and stable gene delivery into human CD4+ T lymphocytes upon in vivo administration of lentiviral vectors, opening an avenue towards novel strategies in immunotherapy.

Ex vivo

Systemic administration

Transduction (biophysics)

10.1016/s1525-0016(16)34175-2

Cite

Citations (0)

Empowering Natural Killer Cells to Combat Acute Myeloid Leukemia: Perspective on CAR-NK Cell Therapy

Transfusion Medicine and Hemotherapy (2024)

F Gierschek J Schlueter Ines Kühnel Frederik Fabian Feigl Dominik Schmiedel

Acute myeloid leukemia (AML) is an aggressive hematologic malignancy with a high relapse rate and still limited therapeutic options. Natural killer (NK) cell-based immunotherapy has the potential to improve outcomes for patients with AML. Recent preclinical studies and early-stage clinical trials aim to enhance the intrinsic anti-leukemic properties of NK cells by selectively targeting AML cells with chimeric antigen receptors (CARs). Furthermore, NK and CAR-NK cells can be combined with other therapeutic modalities or engineered further to overcome the immunosuppressive microenvironment, and treatment resistance of AML blasts and leukemia-initiating cells (LIC). In this review, we summarize preclinical studies with cytokine-stimulated or genetically engineered NK cells derived from different cell sources for the treatment of AML and their translation into early-phase clinical trials. We also provide an overview of promising recent developments toward innovative NK cell-based therapies that may be implemented in the near future.

Cell therapy

10.1159/000540962

Cite

Citations (0)