Engineering NK Cells Modified With an EGFRvIII-specific Chimeric Antigen Receptor to Overexpress CXCR4 Improves Immunotherapy of CXCL12/SDF-1α-secreting Glioblastoma.

Abstract

Natural killer (NK) cells are promising effector cells for adjuvant immunotherapy of cancer. So far, several preclinical studies have shown the feasibility of gene-engineered NK cells, which upon expression of chimeric antigen receptors (CARs) are redirected to otherwise NK cell-resistant tumors. Yet, we reasoned that the efficiency of an immunotherapy using CAR-modified NK cells critically relies on efficient migration to the tumor site and might be improved by the engraftment of a receptor specific for a chemokine released by the tumor. On the basis of the DNAX-activation protein 12 (DAP12), a signaling adapter molecule involved in signal transduction of activating NK cell receptors, we constructed an epidermal growth factor variant III (EGFRvIII)-CAR, designated MR1.1-DAP12 which confers specific cytotoxicity of NK cell towards EGFRvIII glioblastoma cells in vitro and to established subcutaneous U87-MG tumor xenografts. So far, infusion of NK cells with expression of MR1.1-DAP12 caused a moderate but significantly delayed tumor growth and increased median survival time when compared with NK cells transduced with an ITAM-defective CAR. Notably, the further genetic engineering of these EGFRvIII-specific NK cells with the chemokine receptor CXCR4 conferred a specific chemotaxis to CXCL12/SDF-1α secreting U87-MG glioblastoma cells. Moreover, the administration of such NK cells resulted in complete tumor remission in a number of mice and a significantly increased survival when compared with the treatment of xenografts with NK cells expressing only the EGFRvIII-specific CAR or mock control. We conclude that chemokine receptor-engineered NK cells with concomitant expression of a tumor-specific CAR are a promising tool to improve adoptive tumor immunotherapy.

• Authors: Nadja Müller, Susanne Michen, Stefanie Tietze, Katrin Töpfer, Alexander Schulte, Katrin Lamszus, Marc Schmitz, Gabriele Schackert, Ira Pastan, Achim Temme
• Journal: Journal of immunotherapy (Hagerstown, Md. : 1997) (J Immunother) Vol. 38 Issue 5 Pg. 197-210 (Jun 2015) ISSN: 1537-4513 [Electronic] United States
• PMID: 25962108 (Publication Type: Journal Article, Research Support, N.I.H., Intramural, Research Support, Non-U.S. Gov't)

Chemical References

• Chemokine CXCL12
• Receptors, Antigen, T-Cell
• Receptors, CXCR4
• Recombinant Fusion Proteins
• epidermal growth factor receptor VIII
• ErbB Receptors

Topics

• Animals
• Cell Line
• Cell- and Tissue-Based Therapy
• Chemokine CXCL12 (biosynthesis)
• Chemotaxis (genetics, immunology)
• Cytotoxicity, Immunologic
• Disease Models, Animal
• Ectopic Gene Expression
• ErbB Receptors (immunology)
• Female
• Gene Expression
• Gene Order
• Genetic Therapy
• Genetic Vectors (administration & dosage, genetics)
• Glioblastoma (etiology, metabolism, mortality, pathology, therapy)
• Humans
• Immunotherapy
• Killer Cells, Natural (immunology, metabolism)
• Lentivirus (genetics)
• Mice
• Receptors, Antigen, T-Cell (genetics, metabolism)
• Receptors, CXCR4 (genetics, metabolism)
• Recombinant Fusion Proteins (genetics, metabolism)
• Transduction, Genetic
• Tumor Burden
• Xenograft Model Antitumor Assays