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Enhanced antitumor immune response in melanoma tumor model by anti-PD-1 small interference RNA encapsulated in nanoliposomes

Cancer Gene Therapy volume 29, pages 814–824 (2022)Cite this article

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Abstract

Programmed cell death protein-1 (PD-1), as an immune checkpoint molecule, attenuates T-cell activity and induces T-cell exhaustion. Although siRNA has a great potential in cancer immunotherapy, its delivery to target cells is the main limitation of using siRNA. This study aimed to prepare a liposomal formulation as a siRNA carrier to silence PD-1 expression in T cells and investigate it’s in vivo antitumor efficacy. The liposomal siRNA was prepared and characterized by size, zeta potential, and biodistribution. Following that, the uptake assay and mRNA silencing were evaluated in vitro at mRNA and protein levels. siRNA-PD-1 (siPD-1)-loaded liposome nanoparticles were injected into B16F0 tumor-bearing mice to evaluate tumor growth, tumor-infiltrating lymphocytes, and survival rate. Liposomal siPD-1 efficiently silenced PD-1 mRNA expression in T cells (P < 0.0001), and siPD-1-loaded liposomal nanoparticles enhanced the infiltration of T-helper 1 (Th 1) and cytotoxic T lymphocytes into the tumor tissue (P < 0.0001). Liposome-PD-1 siRNA monotherapy and PD-1 siRNA-Doxil (liposomal doxorubicin) combination therapy improved the survival significantly, compared to the control treatment (P < 0.001). Overall, these findings suggest that immunotherapy with siPD-1-loaded liposomes by enhancing T-cell-mediated antitumor immune responses could be considered as a promising strategy for the treatment of melanoma cancer.

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Fig. 1: Stability of liposome in several time points.
Fig. 2: siRNA uptake with lipofectamine and liposome.
Fig. 3: PD-1 protein and mRNA silencing by Anti-PD-1 siRNA.
Fig. 4: In vivo biodistribution of PD-1 siRNA liposome in B16F0 tumor-bearing mice.
Fig. 5: Flow cytometry analysis of tumor-infiltrated lymphocytes.
Fig. 6: Efficacy of PD-1 siRNA on B16F0 melanoma tumor.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

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Funding

We are thankful for the financial support from the Nanotechnology Research, Mashhad University of medical sciences, Mashhad, Iran (grant number: 971960). We would also like to thank Dr. Amir Abbas Momtazi-Borojeni, a post-doctoral researcher of Iran’s National Elites Foundation at the Mashhad University of Medical Sciences, for the final scientific polishing and edition of the present paper.

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Authors and Affiliations

  1. Allergy Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Mehdi Barati

  2. Department of Immunology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Mehdi Barati

  3. Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Farshad Mirzavi & Anvar Soleimani

  4. Molecular Medicine Research Center, Hormozgan Health Institute, Hormozgan University of Medical Sciences, Bandar Abbas, Iran

    Amin Reza Nikpoor

  5. Immunobiochemistry Department, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Mojtaba Sankian

  6. Department of Pathobiology and medical laboratory science, North Khorasan University of Medical Sciences, Bojnurd, Iran

    Hasan Namdar Ahmadabad

  7. Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

    Mohammad Mashreghi & Mahmoud Reza Jaafari

  8. Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

    Jalil Tavakol Afshar & Mojgan Mohammadi

  9. Department of Pharmaceutical Nanotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

    Mahmoud Reza Jaafari

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Contributions

MB conceived the presented idea, wrote the manuscript, performed the experiments, and analyzed data. He also designed the study. FM assisted with measurements. ARN gave consultation. MS gave consultation with the model preparation. HNA helped with the edition of manuscript. AS contributed to sample preparation. MM assisted with measurements. JTA helped with his laboratory facilities. MM supervised the study and gavec onsultation with the writing of the manuscript. MRJ supervised the project and financially supported the study.

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Correspondence to Mojgan Mohammadi or Mahmoud Reza Jaafari.

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Barati, M., Mirzavi, F., Nikpoor, A.R. et al. Enhanced antitumor immune response in melanoma tumor model by anti-PD-1 small interference RNA encapsulated in nanoliposomes. Cancer Gene Ther 29, 814–824 (2022). https://doi.org/10.1038/s41417-021-00367-9

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