Self-Assembled Nanoparticles Prepared from Low-Molecular-Weight PEI and Low-Generation PAMAM for EGFRvIII-Chimeric Antigen Receptor Gene Loading and T-Cell Transient Modification
Received 5 September 2019
Accepted for publication 2 January 2020
Published 23 January 2020 Volume 2020:15 Pages 483—495
Checked for plagiarism Yes
Review by Single anonymous peer review
Peer reviewer comments 2
Editor who approved publication: Dr Mian Wang
Qianru Yu,1,* Maxin Zhang,1,* Yuetan Chen,1,* Xiaolong Chen,2 Sanyuan Shi,1 Kang Sun,1 Ran Ye,1 Yuan Zheng,1 Yang Chen,1 Yuhong Xu,1,3 Jinliang Peng1
1School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, People’s Republic of China; 2NanoDrug Platform, Zhejiang California International NanoSystems Institute, Zhejiang University, Hangzhou 310058, People’s Republic of China; 3School of Pharmacy and Chemistry, Dali University, Dali City 671000, People’s Republic of China
*These authors contributed equally to this work
Correspondence: Jinliang Peng
School of Pharmacy, Shanghai Jiao Tong University, No. 800, Dongchuan Road, Shanghai 200240, People’s Republic of China
Tel/Fax +86 21 3420 4739
Background: The complex preparation procedures and severe toxicities are two major obstacles facing the wide use of chimeric antigen receptor-modified T (CAR-T) cells in clinical cancer immunotherapy. The nanotechnology-based T cell temporary CAR modification may be a potential approach to solve these problems and make the CAR-T cell-based tumor therapy feasible and broadly applicable.
Methods: A series of plasmid DNA-loaded self-assembled nanoparticles (pDNA@SNPsx/y) prepared from adamantane-grafted polyamidoamine (Ad-PAMAM) dendrimers of different generations (G1 or G5) and cyclodextrin-grafted branched polyethylenimine (CD-PEI) of different molecular weights (800, 2000, or 25,000 Da) were characterized and evaluated. The detailed physicochemical properties, cellular interaction, and cytotoxicity of selected pDNA@SNPG1/800 were systematically investigated. Thereafter, the epidermal growth factor receptor variant III (EGFRvIII) CAR-expression plasmid vector (pEGFRvIII-CAR) was constructed and encapsulated into SNPG1/800. The resulting pEGFRvIII-CAR@SNPG1/800 was used for Jurkat cell transient transfection, and the EGFRvIII-CAR expressed in transfected cells was measured by flow cytometry and Western blot. Finally, the response of EGFRvIII CAR-positive Jurkat T cell to target tumor cell was evaluated.
Results: The pDNA@SNPG1/800 showed the highest efficacy in Jurkat cell gene transfection and exhibited low cytotoxicity. pEGFRvIII-CAR@SNPG1/800 can efficiently deliver pEGFRvIII-CAR into Jurkat T cells, thereby resulting in transient EGFRvIII-CAR expression in transfected cells. EGFRvIII-CAR that is present on the cell membrane enabled Jurkat T cells to recognize and bind specifically with EGFRvIII-positive tumor cells.
Conclusion: These results indicated that pEGFRvIII-CAR@SNPG1/800 can effectively achieve T-cell transient CAR modification, thereby demonstrating considerable potential in CAR-T cancer therapy.
Keywords: self-assembled nanoparticle, PAMAM dendrimer, polyethylenimine, epidermal growth factor receptor variant III, chimeric antigen receptor, T lymphocyte