Hit-and-run programming of therapeutic cytoreagents using mRNA nanocarriers

Abstract

Therapies based on immune cells have been applied for diseases ranging from cancer to diabetes. However, the viral and electroporation methods used to create cytoreagents are complex and expensive. Consequently, we develop targeted mRNA nanocarriers that are simply mixed with cells to reprogram them via transient expression. Here, we describe three examples to establish that the approach is simple and generalizable. First, we demonstrate that nanocarriers delivering mRNA encoding a genome-editing agent can efficiently knock-out selected genes in anti-cancer T-cells. Second, we imprint a long-lived phenotype exhibiting improved antitumor activities into T-cells by transfecting them with mRNAs that encode a key transcription factor of memory formation. Third, we show how mRNA nanocarriers can program hematopoietic stem cells with improved self-renewal properties. The simplicity of the approach contrasts with the complex protocols currently used to program therapeutic cells, so our methods will likely facilitate manufacturing of cytoreagents.

Polymeric Nonviral Gene Delivery Systems for Cancer Immunotherapy

“…successfully designed a new CAR‐T preparation method which delivered mRNA‐coated nanoparticle into stem cells or T cells from the patients in vitro (Figure 10b). [ 104 ]…”

Nanotechnology Promotes Genetic and Functional Modifications of Therapeutic T Cells Against Cancer

“…[ 71 ] Based on this concept, Moffett et al effectively removed the T cell receptor alpha constant region (TRAC) using megaTAL nuclease and abolished the production of non‐cancer‐specific T cell receptors (TCRs), which is an important challenge in improving CAR‐T cell therapy. [ 63 ] To achieve their goal, they delivered mRNA genes to T cells by targeting CD3‐ and CD8‐cells. Their results confirmed that the addition of bioengineered NPs can serve to target specific cell subtypes, stimulate receptor‐mediated endocytosis (namely, the “enhancement of RNA entry by a physiological process without compromising cell viability”), transiently program the expression of genes, and improve the therapeutic potential of both programmed T cells and stem cells.…”

“…Given the essential role of cancer‐associated cells in tumor progression and metastasis, it is not surprising that targeting lymphocytes or hematopoietic stem cells (HSCs) with NPs has proven to be an exciting therapeutic option. [ 63 ] Using this concept, NP‐based gene delivery to tumor‐associated cells can induce the transformation of the immunosuppressive TME into a toxic environment for cancer cells, [ 106 ] which may potentially offer a universal approach to immunotherapy. [ 107 ] Furthermore, NPs appear to be a promising tool to reprogram tumor‐associated cells such as fibroblasts, immune cells (macrophages), dendritic cells (DCs), blood endothelial cells (ECs), and lymphatic vasculators.…”

“…Within this framework, the unique characteristics of NPs (size, shape, surface moieties) have been explored extensively to achieve superior conditions improving T cell programming, transfection, and therapeutic efficiency. [ 63,73 ] Representative examples of such NP systems are summarized in Table 3. It should be considered that nucleic acids comprise either ribose (RNA) or deoxyribose (DNA) and display additional properties such as hydrophilicity and negative charges allowing them to be efficiently encapsulated into NPs.…”

Realizing Cancer Precision Medicine by Integrating Systems Biology and Nanomaterial Engineering

“…[73][74][75][76] Moffett et al and Smith et al each developed a nanocarrier that specifically targets the T cells of patients and delivers mRNA or a DNA plasmid into the T cells to transform them into chimeric antigen receptor (CAR) T cells in situ or ex vivo. [77,78] In this nanocarrier, mRNA or a DNA plasmid encoding the CAR is condensed by incubation with poly(beta-amino ester) (PBAE) 447 polymer and a polyglutamic acid-antibody recognizing cluster of differentiation 3 (CD3), a protein found on the surface of T cells. Through the CD3-specific antibody, the nanocarrier interacts with T cells.…”