siRNA Nanoparticle Targeting PD-L1 Activates Tumor Immunity and Abrogates Pancreatic Cancer Growth in Humanized Preclinical Model
Abstract
:1. Introduction
2. Materials and Methods
2.1. Synthesis of siPD-L1@PLGA NPs
2.2. Derivation of Primary Pancreatic Cancer Cell and Humanized PDX Model
2.3. Cell Culture and FACS
2.4. Antibodies and Reagents
2.5. Cellular Uptake of siPD-L1@PLGA NPs
2.6. Cytotoxicity Study of Scrambled siPD-L1@PLGA NPs on Blue #96 Cells
2.7. Isolation of Splenocytes and CD8+ T cells
2.8. In Vitro Cytolytic Assay of Ovalbumin(OVA)-Specific Cytotoxic T Lymphocytes (CTLs)
2.9. In Vitro Proliferation Study of OVA-Specific CTLs
2.10. Production of IFN-γ in Tumor Antigen-Stimulated CTLs
2.11. siPD-L1@PLGA NPs Treatment and Analysis of Tumor-Infiltrated Immune Cells in Humanized NSG Model
2.12. Measurement of Lymphocyte-Mediated Cytotoxicity from Tumor-Bearing Mouse
2.13. Multiplexed Fluorescent Immunohistochemistry
2.14. Multispectral Imaging and Analysis
2.15. DLS Analysis of siRNA@PLGA NPs
2.16. Statistics
3. Results
3.1. Synthesis of siRNA Nanoparticles Targeting PD-L1 and In Vitro Validation
3.2. siPD-L1@PLGA Abrogates Immune Escape Function of Pancreatic Tumor Cells
3.3. Efficacy Test in Humanized PDX Model Reveals Antitumor Effect of siPD-L1@PLGA via Immunomodulation
3.4. OPAL Multiplex IHC Analysis Reveals Granzyme B Upregulation in siPD-L1@PLGA-Treated Tumors
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Jung, J.Y.; Ryu, H.J.; Lee, S.-H.; Kim, D.-Y.; Kim, M.J.; Lee, E.J.; Ryu, Y.-M.; Kim, S.-Y.; Kim, K.-P.; Choi, E.Y.; et al. siRNA Nanoparticle Targeting PD-L1 Activates Tumor Immunity and Abrogates Pancreatic Cancer Growth in Humanized Preclinical Model. Cells 2021, 10, 2734. https://doi.org/10.3390/cells10102734
Jung JY, Ryu HJ, Lee S-H, Kim D-Y, Kim MJ, Lee EJ, Ryu Y-M, Kim S-Y, Kim K-P, Choi EY, et al. siRNA Nanoparticle Targeting PD-L1 Activates Tumor Immunity and Abrogates Pancreatic Cancer Growth in Humanized Preclinical Model. Cells. 2021; 10(10):2734. https://doi.org/10.3390/cells10102734
Chicago/Turabian StyleJung, Jae Yun, Hyun Jin Ryu, Seung-Hwan Lee, Dong-Young Kim, Myung Ji Kim, Eun Ji Lee, Yeon-Mi Ryu, Sang-Yeob Kim, Kyu-Pyo Kim, Eun Young Choi, and et al. 2021. "siRNA Nanoparticle Targeting PD-L1 Activates Tumor Immunity and Abrogates Pancreatic Cancer Growth in Humanized Preclinical Model" Cells 10, no. 10: 2734. https://doi.org/10.3390/cells10102734