Nanoparticle-Based Chimeric Antigen Receptor Therapy for Cancer Immunotherapy

Abstract

Adoptive cell therapy with chimeric antigen receptor (CAR)-engineered T cells (CAR-Ts) has emerged as an innovative immunotherapy for hematological cancer treatment. However, the limited effect on solid tumors, complex processes, and excessive manufacturing costs remain as limitations of CAR-T therapy. Nanotechnology provides an alternative to the conventional CAR-T therapy. Owing to their unique physicochemical properties, nanoparticles can not only serve as a delivery platform for drugs but also target specific cells. Nanoparticle-based CAR therapy can be applied not only to T cells but also to CAR-natural killer and CAR-macrophage, compensating for some of their limitations. This review focuses on the introduction of nanoparticle-based advanced CAR immune cell therapy and future perspectives on immune cell reprogramming.

Keywords: Cancer immunotherapy; Chimeric antigen receptor (CAR); Genetic engineering; Immune cell reprograming; Nanoparticle.

Fig. 1

A Schematic illustration of CAR and CAR-T and B Schematic diagram of conventional process of CAR-T therapy using viral vectors (created with BioRender.com)

Fig. 2

A Schematic diagram of preparation and formulation screening of SNPs by comparing T cell transfection efficiency depending on the type of Ad-PAMAM and CD-PEI, and anticancer effect of anti-EGFRvIII CAR-T. B Transfection efficiency of Lipofectamine 2000 (Lipo2000), PEI800, or SNPG1/800 via luciferase activity (*p < 0.05). C Confocal microscopy images of interaction between EGFRvIII-positive HuH7 cells (green) and Jurkat T cells (red) transfected with or without pEGFRvIII-CAR@SNPs^G1/800. Scale bars, 20 μm. (Reproduced from a previous report [29] with permission from Dove Medical Press Limited)

Fig. 3

A Schematic diagram of LNP preparation via microfluidic device. B Viability of primary T cells treated with crude LNP, pure LNP, and EP group. n = 3 biological replicates, *p < 0.05 in paired t-test to EP. C Comparison of tumor cell killing ability in vitro of CAR-Ts engineered by EP, LNP (C14-4), lentiviral vector, and control. n = 3 wells. *p < 0.05, **p < 0.01 in the paired t-test to control. (Reproduced from a previous report [30] with permission from the American Chemical Society)

Fig. 4

A Schematic diagram of fabrication of anti-CD3e f(ab′)2-coated PBAE NP carrying CAR plasmid DNA for in vivo CAR generation. B Flow cytometry of peripheral T cells after injection of NP encapsulating 194-1BBz_2A_GFP genes. C Survival of animals following the various groups of CAR-T therapy. (Reproduced from a previous report [31] with permission from Nature Publishing Group)

Fig. 5

A Schematic illustration of T cell engineering via FAP CAR-mRNA/LNP against activated fibroblast. B Percentage of FAPCAR positive T cells isolated from mice 48 h after injection of 10 μg of mRNA-LNPs. C Histologic analysis of coronal cardiac sections of animals and quantification of fibrosis. Picrosirius red staining indicates collagen (pink). Inset shows magnification of the LV myocardium. Scale bar, 100 μm. (Reproduced from a previous report [32] with permission from American Association for the Advancement of Science)

Fig. 6

A Schematic diagram of CART/mRNA polyplex preparation. B Percentage of antihuman CD19-41BB-CD3ζ CAR expression by isolated primary resting human NK cells. C Percentage of dead target cells (CD19 positive Nalm6: filled circles, CD19 knockout Nalm6: open circles) after 20:1 (transfected NK:target) ratio coculture for 6 h before flow cytometric analysis. (Reproduced from a previous report [33] with permission from the American Society of Hematology)

Fig. 7

A Schematic illustration of multifunctional nanoparticle (MF-NP) and its bioapplications. B Quantitative analysis data of EGFR-CAR expression on NK-92MI surface and cancer cell killing capability of EGFR-CAR expressing NK-92MI cells according to the concentration of MF-NPs. in vivo fluorescence, optical imaging C and MR imaging D of mice transplanted with MF-NP–labeled NK-92MI cells. (Reproduced from a previous report [34] with permission from Elsevier)

Fig. 8

A Schematic diagram of MPEI/pDNA(CAR-IFN-γ) nanocomplex preparation and antitumoral mechanisms of CAR-M1 macrophages. B Representative fluorescence images (left) of major organs and tumor 48 h after an intra-tumoral injection of vehicle (negative control) or MPEI/pCAR-IFN-γ. Flow cytometry data (right) of CAR + cell percentages in the tumor tissues (n = 3). C Mean tumor growth profiles of Neuro-2a tumor-bearing mice after various treatments (n = 15). D Survival rate of Neuro-2a-bearing mice after various treatments (n = 6). (Reproduced from a previous report [35] with permission from Wiley‐VCH)

Fig. 9

A Schematic illustration of CAR mRNA delivery to murine primary macrophages and killing assay of FLuc⁺ human B lymphoma. B Flow cytometry data of transfection efficiency of m1ψ-eGFP mRNA in Mφ and M1 treated in different formulations of 9322-O16B. C Killing assay of FLuc⁺ B lymphoma following different treatments (Mφ, p < 0.05*; M1, p < 0.05*). Data are presented as mean ± SD (n = 3). (Reproduced from a previous report [36] with permission from the American Chemical Society)