Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis
1
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.
Abstract
The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.
참고문헌 (31)
- Leslie M, "New cancer-fighting cells enter trials", Science, 361, pp.1056~1057 (2018)
- Overdijk MB, "Antibody-mediated phagocytosis contributes to the anti-tumor activity of the therapeutic antibody daratumumab in lymphoma and multiple myeloma", MAbs, 7, pp.311~321 (2015)
- Chao MP, "Anti-CD47antibody synergizes with rituximab to promote phagocytosis and eradicate non-Hodgkin lymphoma", Cell, 142, pp.699~713 (2010)
- Shi Y, "Trastuzumab triggers phagocytic killing of high HER2 cancer cells in vitro and in vivo by interaction with Fcgamma receptors on macrophages", J Immunol, 194, pp.4379~4386 (2015)
- Kurdi AT, "Antibody-Dependent Cellular Phagocytosis by Macrophages is a Novel Mechanism of Action of Elotuzumab", Mol Cancer Ther, 17, pp.1454~1463 (2018)
- Michaud HA, "Tumor antigen-targeting monoclonal antibody-based immunotherapy : Orchestrating combined strategies for the development of long-term antitumor immunity", Oncoimmunology, 3, pp.e955684~ (2014)
- Lee S, "Macrophage-based cell therapies : The long and winding road", J Control Release, 240, pp.527~540 (2016)
- Genard G, "Reprogramming of Tumor-Associated Macrophages with Anticancer Therapies : Radiotherapy versus Chemo-and Immunotherapies", Front Immunol, 8, pp.828~ (2017)
- Engelman A, "The ups and downs of gene expression and retroviral DNA integration", Proc Natl Acad Sci U S A, 102, pp.1275~1276 (2005)
- Shayakhmetov DM, "Adenovirus binding to blood factors results in liver cell infection and hepatotoxicity", J Virol, 79, pp.7478~7491 (2005)
- Zhang X, "The expression of exogenous genes in macrophages : obstacles and opportunities", Methods Mol Biol, 531, pp.123~143 (2009)
- Hardee CL, "Advances in Non-Viral DNA Vectors for Gene Therapy", Genes (Basel), 8, pp.65~ (2017)
- Mun JY, "Minicircle microporation-based non-viral gene delivery improved the targeting of mesenchymal stem cells to an injury site", Biomaterials, 101, pp.310~320 (2016)
- Latz E, "TLR9signals after translocating from the ER to CpG DNA in the lysosome", Nat Immunol, 5, pp.190~198 (2004)
- Hyde SC, "CpG-free plasmids confer reduced inflammation and sustained pulmonary gene expression", Nat Biotechnol, 26, pp.549~551 (2008)
- Lesina E, "CpG-free plasmid DNA prevents deterioration of pulmonary function in mice", Eur J Pharm Biopharm, 74, pp.427~434 (2010)
- Lesina E, "The effect of CpG motifs on gene expression and clearance kinetics of aerosol administered polyethylenimine(PEI)-plasmid DNA complexes in the lung", J Control Release, 143, pp.243~250 (2010)
- Pringle IA, "CpG-free plasmid expression cassettes for cystic fibrosis gene therapy", Biomaterials, 33, pp.6833~6842 (2012)
- Mann CJ, "Molecular signature of the immune and tissue response to non-coding plasmid DNA in skeletal muscle after electrotransfer", Gene Ther, 19, pp.1177~1186 (2012)
- Yew NS, "CpG-depleted plasmid DNA vectors with enhanced safety and long-term gene expression in vivo", Mol Ther, 5, pp.731~738 (2002)
- Hodges BL, "Long-term transgene expression from plasmid DNA gene therapy vectors is negatively affected by CpG dinucleotides", Mol Ther, 10, pp.269~278 (2004)
- Morrissey MA, "Chimeric antigen receptors that trigger phagocytosis", eLife, 7, pp.e36688~ (2018)
- Mosaad YM, "Hematopoietic stem cells : an overview", Transfus Apher Sci, 51, pp.68~82 (2014)
- Piechaczek C, "A vector based on the SV40 origin of replication and chromosomal S", MARs replicates episomally in CHO cells, 1999, pp.27, 426~428
- Ramezani A, "Performance-and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator", Blood, 101, pp.4717~4724 (2003)
- Schubeler D, "Scaffold", matrix-attached regions act upon transcription in a context-dependent manner, 1996, pp.35, 11160~11169
- Giannakopoulos A, "The functional role of S", MARs in episomal vectors as defined by the stressinduced destabilization profile of the vector sequences, 2009, pp.387, 1239~1249
- Stacey KJ, "Electroporation and DNA-dependent cell death in murine macrophages", Immunol Cell Biol, 71(Pt 2), pp.75~85 (1993)
- Rupprecht AP, "Transfection of adherent murine peritoneal macrophages with a reporter gene using DEAE-dextran", J Immunol Methods, 144, pp.157~163 (1991)
- Thompson CD, "Evaluation of methods for transient transfection of a murine macrophage cell line, RAW 264.7", Biotechniques, 27, pp.824~826 (1999)
- Shin DM, "Mycobacterium abscessus activates the macrophage innate immune response via a physical and functional interaction between TLR2and dectin-1", Cell Microbiol, 10, pp.1608~1621 (2008)
