<p>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</p>

Abstract

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 CART cell-based tumor therapy feasible and broadly applicable. Methods: A series of plasmid DNA-loaded self-assembled nanoparticles (pDNA@SNPs x/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@SNP G1/800 were systematically investigated. Thereafter, the epidermal growth factor receptor variant III (EGFRvIII) CAR-expression plasmid vector (pEGFRvIII-CAR) was constructed and encapsulated into SNP G1/800. The resulting pEGFRvIII-CAR@SNP G1/800 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@SNP G1/800 showed the highest efficacy in Jurkat cell gene transfection and exhibited low cytotoxicity. pEGFRvIII-CAR@SNP G1/800 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@SNP G1/800 can effectively achieve T-cell transient CAR modification, thereby demonstrating considerable potential in CART cancer therapy.

Non-viral Vectors in Gene Therapy: Recent Development, Challenges, and Prospects

“…have demonstrated that self-assembled nanoparticles (SNPs) prepared from cyclodextrin-grafted low molecular weight PEI (CD-PEI800) presented low cytotoxicity and a high transfection efficiency to Jurkat cells. They postulated that the cationic hydrogel generated from CD-PEI800 contributed to SNPs’ enhanced gene encapsulation efficiency ( 64 ).…”

Recent Advances in Preclinical Research Using PAMAM Dendrimers for Cancer Gene Therapy

“…The complex successfully delivered the plasmid with EGFRvIII-CAR to Jurkat T cells, which specifically recognized EGFRvIII-positive cancer cells ( Figure 7 ). This study has demonstrated the high transient CAR transfection efficiency and low toxicity towards Jurkat T cells, which, in the view of the research team, can be used in future in vivo and ex vivo studies of the given complex, forming a basis for potential clinical trials [ 153 ].…”

“…Abbreviations: pDNA, plasmid DNA; Ad-PEG, adamantane-grafted poly(ethylene glycol); Ad-PAMAMx, adamantane-grafted polyamidoamine dendrimer (x: PAMAM dendrimer generation); CD-PEIy, cyclodextrin-grafted branched polyethylenimine (y: PEI molecular weight); SNPs, self-assembled nanoparticles; EGFRvIII, epidermal growth factor receptor variant III; CAR, chimeric antigen receptor; pEGFRvIII-CAR, epidermal growth factor receptor variant III-chimeric antigen receptor expression plasmid. With the permission of [ 153 ], copyright (2021) Dove Medical Press.…”

Nanocarriers Used in Drug Delivery to Enhance Immune System in Cancer Therapy

“…The results of the study showed the ability of LNPs to deliver mRNA from the CAR to primary T cells and generate functional CAR T cells that have the potential to induce cancer cell death [ 159 ]. Furthermore, another approach to reprogramming T cells is to use nanocarriers to deliver drugs, antibodies, and interleukins, as they can protect T cells from immunosuppression and activate CD8+ T cells [ 160 , 161 , 162 , 163 , 164 ].…”