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SCIENCE CHINA Materials, Volume 64 , Issue 8 : 2045-2055(2021) https://doi.org/10.1007/s40843-020-1591-1

Gene silencing-mediated immune checkpoint blockade for tumor therapy boosted by dendrimer-entrapped gold nanoparticles

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  • ReceivedOct 19, 2020
  • AcceptedDec 14, 2020
  • PublishedMar 5, 2021

Abstract


Funding

the National Key R&D Program of China(2017YFE0196200)

the National Natural Science Foundation of China(81761148028,21773026)

and the Science and Technology Commission of Shanghai Municipality(19XD1400100,205207130300,19410740200)


Acknowledgment

This work was supported by the National Key R&D Program of China (2017YFE0196200), the National Natural Science Foundation of China (81761148028 and 21773026), and the Science and Technology Commission of Shanghai Municipality (19XD1400100, 205207130300, 20DZ2254900 and 19410740200). Shi X also acknowledges the support by FCT-Fundação para a Ciência e a Tecnologia through the CQM Base Fund—UIDB/00674/2020, and Programmatic Fund—UIDP/00674/2020, and by ARDITI-Agência Regional para o Desenvolvimento da Investigação Tecnologia e Inovação, through the project M1420-01-0145-FEDER-000005—Centro de Química da Madeira—CQM+ (Madeira 14-20 Program).


Interest statement

The authors declare no competing financial interest.


Contributions statement

Xue X carried out the experiments including synthesis, characterization, and in vitro and in vivo evaluation of the vector/siPD-L1 polyplexes, and wrote the original draft of the manuscript; Li J and Fan Y carried out part of the material characterization and in vitro evaluation of the materials; Shen M performed part of the data analysis and interpretation. Shi X designed and supervised the whole project and did the final manuscript editing.


Author information

Xue Xue obtained her BS degree in pharmaceutical engineering from Shanghai University of Engineering Science in 2018. Now she is an MS student of biomedical engineering at Donghua University under the supervision of Prof. Xiang-yang Shi. Her research interests focus on the design of dendrimer-based systems for cancer therapy.


Xiangyang Shi obtained his PhD degree in 1998 from the Chinese Academy of Sciences. From 2002 to 2008, he was appointed as a research fellow, research associate II, research investigator, and research assistant professor at the University of Michigan, Ann Arbor. In September 2008, he joined Donghua University as a full professor. His research interests focus on organic/inorganic hybrid nanoplatforms for sensing, imaging, and theranostic applications, in particular for precision cancer imaging and therapy.


Supplementary data

Supplementary information

Supporting data are available in the online version of the paper.


References

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  • Figure 1

    Preparation of {(Au0)25-G5.NH2-mPEG-F} DENPs/siPD-L1 polyplexes for ICB-based cancer immunotherapy.

  • Figure 2

    (a) TEM and magnified TEM (inset) images of the {(Au0)25-G5.NH2-mPEG-F} DENPs. (b) Gel retardation assay of siPD-L1 after complexing with the vector at different N/P ratios. Lanes 1–8 represent naked siRNA and N/P ratios of 0.125:1, 0.25:1, 0.5:1, 1:1, 2:1, 3:1 and 4:1, respectively. Zeta potential (c) and hydrodynamic size (d) of vector/siPD-L1 polyplexes under different N/P ratios.

  • Figure 3

    (a) Cytotoxicity assay of vector/siPD-L1 and vector/siNC polyplexes after B16 cells were treated with them at different vector concentrations. (b) Relative mean fluorescence of B16 cells transfected with vector/siPD-L1 polyplexes at N/P ratios of 2:1, 5:1, 10:1, and 20:1, respectively, for 4 h was analyzed by flow cytometry. The fluorescence intensity of PBS group was set to be 1.0 (mean ± SD, n = 3). (c) Western blot assay of PD-L1 protein expression (mean ± SD, n = 3). Expression of PD-L1 was set to be 1.0 in cells treated with PBS, and *** represents p < 0.001. (d) Confocal microscopic imaging of B16 cells transfected with PBS, free siPD-L1 or vector/siPD-L1 (N/P value = 10) for 4 h. The scale bar represents 20 µm for each panel.

  • Figure 4

    (a) The timeline used for the tumor immunotherapy in vivo. (b) The relative tumor volume changes of the mice in different treatment groups (n = 6 for each group, and ** and *** represent p < 0.01 and p < 0.001, respectively). (c) The relative mouse body weight variations in different treatment groups (n = 6 for each group). (d) H&E and TUNEL staining of the tumor sections on the 14th day after different treatments. Scale bar represents 100 µm for each panel.

  • Figure 5

    (a) Western blot assay of PD-L1 expression of tumor cells. Expression of PD-L1 in tumor cells treated with PBS was set to be 1.0. (b) The immunofluorescence images of PD-L1, CD4 and CD8 proteins in tumor tissues. (c–e) Flow cytometric assay of CD4+/CD8+ T cells in the infiltrating tumor tissues (gated on CD3+ T cells). (f–h) Contents of the TNF-α, IFN-γ and IL-6 in serum on the 14th day after various treatments. All data are shown as mean ± SD (n = 3). In (a) and (d–h), ** and *** represent p < 0.01 and p < 0.001, respectively.

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