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Modification of the tumor microenvironment enhances immunity with plasmid gene therapy

Abstract

Local intratumor delivery with electroporation of low levels of plasmids encoding molecules, induces an antitumor effect without causing systemic toxicity. However, previous studies have predominately focused on the function of the delivered molecule encoded within the plasmid, and ignored the plasmid vector. In this study, we found vectors pUMVC3 and pVax1 induced upregulation of MHC class I (MHC-I) and PD-L1 on tumor cell surface. These molecules participate in a considerable number of immunoregulatory functions through their interactions with and activating inhibitory immune cell receptors. MHC molecules are well-known for their role in antigen (cross-) presentation, thereby functioning as key players in the communication between immune cells and tumor cells. Increased PD-L1 expression on tumor cells is an important monitor of tumor growth and the effectiveness of immune inhibitor therapy. Results from flow cytometry confirmed increased expression of MHC-I and PDL-1 on B16F10, 4T1, and KPC tumor cell lines. Preliminary animal data from tumor-bearing models, B16F10 melanoma, 4T1 breast cancer and KPC pancreatic cancer mouse models showed that tumor growth was attenuated after pUMVC3 intratumoral electroporation. Our data also documented that pSTAT1 signaling pathway might not be associated with plasmid vectors’ function of upregulating MHC-I, PD-L1 on tumor cells.

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Fig. 1: Plasmid vector EP attenuate of established tumor growth.
Fig. 2: MHC-I and PD-L1 surface expression on tumor cells were elevated after plasmid DNA electroporation.
Fig. 3: IFN-γ not IL-12 induces dose-dependent MHC-I and PD-L1 upregulation expression in three different cell lines.
Fig. 4: Regulating surface expression of MHC-I and PD-L1 in cell lines.
Fig. 5: Plasmid DNA upregulation of MHC-I and PD-L1 expression was independent in STAT1 signaling pathway.

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All the data associated with this study are present in the paper or the Supplementary Materials. Materials are available upon request from the corresponding author.

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Acknowledgements

This research was funded in part by the U.S. National Institutes of Health, R01 CA186730. The funders had no role in study design, collection of data, decision to publish, or in preparation of this manuscript.

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RH conceived and supervised the study and aided in interpretation of data. GS and JS performed experiments. GS designed, interpreted, analyzed, and interpreted data.

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Correspondence to Richard Heller.

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Dr. R. Heller is an inventor on patents which cover the technology that was used in the work reported in this manuscript. In addition, Dr. R. Heller owns stock in Inovio Pharmaceuticals, Inc.

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All animal experiments were approved by the University of South Florida Institutional Animal Care and Use Committee (IACUC; protocols #IS00007033, #IS00011648).

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Shi, G., Synowiec, J., Singh, J. et al. Modification of the tumor microenvironment enhances immunity with plasmid gene therapy. Cancer Gene Ther 31, 641–648 (2024). https://doi.org/10.1038/s41417-024-00728-0

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