Immunoengineering via Chimeric Antigen Receptor-T Cell Therapy: Reprogramming Nanodrug Delivery

Abstract

Following its therapeutic effect in hematological metastasis, chimeric antigen receptor (CAR) T cell therapy has gained a great deal of attention during the last years. However, the effectiveness of this treatment has been hampered by a number of challenges, including significant toxicities, difficult access to tumor locations, inadequate therapeutic persistence, and manufacturing problems. Developing novel techniques to produce effective CARs, administer them, and monitor their anti-tumor activity in CAR-T cell treatment is undoubtedly necessary. Exploiting the advantages of nanotechnology may possibly be a useful strategy to increase the efficacy of CAR-T cell treatment. This study outlines the current drawbacks of CAR-T immunotherapy and identifies promising developments and significant benefits of using nanotechnology in order to introduce CAR transgene motifs into primary T cells, promote T cell expansion, enhance T cell trafficking, promote intrinsic T cell activity and rewire the immunosuppressive cellular and vascular microenvironments. Therefore, the development of powerful CART cells can be made possible with genetic and functional alterations supported by nanotechnology. In this review, we discuss the innovative and possible uses of nanotechnology for clinical translation, including the delivery, engineering, execution, and modulation of immune functions to enhance and optimize the anti-tumor efficacy of CAR-T cell treatment.

Keywords: CAR T cell; drug delivery; immunoengineering; reprogramming.

Figure 1

The evolution and structure of MHC-independent CARs. Monoclonal antibodies and T cell receptor complexes are the sources of CAR, forming the ectodomain of the CAR by linking the hinge domains (HD) and combining the VH and VL of the antibody (ScFv). The entire CAR is anchored to the donor or autologous T cell membrane via the transmembrane domain. The CD3-chain, which is the first-generation CAR and contains three ITAMs, is the main source of the intracellular signaling domain. In addition, each of the CD3ε-chains, carries one ITAM. The CD28 CAR of the second generation introduces a co-stimulatory signal into T cells. The T cells of the third-generation CAR (4-1BB and CD28 CAR) integrate two unique co-stimulatory signals. The fourth-generation CAR incorporates cytokine signaling into T cells as well as a co-stimulatory signal. The fifth generation of CAR has modified the intracellular binding sites for STAT3 transcription factor and another attachment region for theIL-2 receptor. Reproduced with permission from Ref. [22]. Copyright 2021, Elsevier Ltd.

Figure 2

Creation and delivery of CAR mRNApolymeric nanoparticles with tumor specificity. (A) Bioengineered polymeric NPs are effectively able to bind to T cells via targeting ligands (Ab). (B) Upon their infusion, there is a brief programming for the expression of CARs that are specific to the tumor. Reproduced with permission from Ref. [37]. Copyright 2020, Springer Nature.

Figure 3

MPEI/pCAR-IFN nanocomplex therapeutic mechanisms. (A) Diagram showing the delivery of gene combination encoding an ALK-specific CAR and an IFN-plasmid DNA (pCAR-IFN) via MPEI to activate CAR-M1 macrophages in vivo and their anti-tumor properties. (B) Transgene construct including both an anti-ALK CAR and an IFN gene. Reproduced with permission from Ref. [49]. Copyright 2021, Wiley.

Figure 4

mRNA nanoparticle production for therapeutic T cell programming. (A) Creating specifically aimed mRNA-carrying NPs. A representative NP is depicted in the transmission electron micrograph inset. The designed synthetic mRNA contained within the NP, which encodes therapeutically important proteins, is also shown. (B) Schematic illustration of the programming of farmed T cells to express transgenes on polymeric nanoparticles (NPs) that are therapeutically used in patients. In order to introduce their mRNA cargoes and cause the targeted cells to express specified proteins (transcription factors or genome-editing agents), these particles are coated with ligands that direct them to particular cell types. Reproduced with permission from Ref. [56]. Copyright 2017, Springer Nature.

Figure 5

Injectable hydrogels triggering local inflammatory niche for the co-delivery of CART cells and cytokines in solid tumor therapy. (A) Schematic illustration comparing the intravenous (IV) administration technique for CART cells to solid tumors to existing methods. (B) The B7H3 CAR construct used in method is shown. (C) The amount of transduction that B7H3 CART cells were able to complete in comparison to untransduced “Mock” T cells after being stained with B7H3-Fc. (D) Dodecyl-modified hydroxypropyl methylcellulose (HPMC) and degradable block-copolymer nanoparticles self-assemble to form PNP hydrogels that co-encapsulate CART cells and stimulate cytokines. Reproduced with permission from Ref. [70]. Copyright 2022, AAAS.

Figure 6

Design and production of lymphocyte-programming nanoparticles. (A) Schematic illustration of the T cell-targeted DNA nanocarrier. A transmission electron micrograph of a typical nanoparticle is shown. The two plasmids that were included in the nanoparticles are also shown; they contain the hyperactive iPB7 transposase and an all-murine 194-1BBz CAR. (B) Diagram describing the fabrication of the poly(β-amino ester) nanoparticles. Reproduced with permission from Ref. [37]. Copyright 2017, Springer Nature.

Figure 7

Combining CAR-T cell therapy with CRISPR method. Using donor T cells with base editing to combat T cell leukemia. Cytidine deamination gives the opportunity for highly targeted CT conversion, the addition of stop codons, or the elimination of splice sites, all of which can be used to impair gene expression without resulting in double-strand DNA breakage. By electroporating three sgRNAs against TRBC, CD7, and CD52 along with mRNA expressing codon-optimized BE3, BE-CAR7 T cells were produced from healthy donor peripheral-blood lymphocytes. This procedure made it possible to target CD7⁺ leukemia cells specifically and express CAR7 following lentiviral transduction without causing fratricide. Reproduced with permission from Ref. [81]. Copyright 2023, NEJM.