Custom T Cell Reprogramming by mRNA

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  • Custom T Cell Reprogramming by mRNA

    T Cells Reprogramming mRNA reprogramming is a “footprint-free” method that can evoke the production of induced pluripotent stem cells (iPSCs) without heritable changes to cellular DNA and an unacceptable risk of tumorigenicity. mRNA-based reprogramming method also contributes to the generation of iPSCs derived from T cells. As an excellent service provider, Creative Biolabs has developed a non-integrative RNA reprogramming platform to produce and deliver high-quality engineered T cells for cell immunotherapy.

    Overview of T Cells Reprogramming

    T cells play crucial roles in mounting effective immune responses that affect therapeutic interventions in cancer, infectious disease, and autoimmunity. It has been revealed that T cells can be programmed with molecular modules to increase their therapeutic potency and specificity, especially in adoptive cell therapy (ACT) for cancers. Genetically reprogrammed T cells hold great promise in the areas of ACT, offering great hope for curative responses in cancer patients.

    • Engineered TCRs
    • T cell receptor (TCR) is sufficient to direct antigen-specific T cell differentiation and redirect their cytotoxicity. Now, TCRs can be engineered to possess more specificity, affinity, reactivity and broad-spectrum binding ability. For instance, TCRs can be engineered to potentially recognize all peptides processed and presented in the context of MHC molecules, thus allowing TCRs to target both surface and intracellular antigens. Exodomains of α and β subunits of the TCR can be modified by replacing their variable domains with antibody domains that can recognize cancer-associated antigens.

    • Chimeric Antigen Receptors (CARs)
    • CARs are a class of synthetic TCR receptors that reprogram lymphocyte specificity and function. It is an effective genetic optimization of T cells to redirect specificity. For instance, CD28-based CARs can promote T cell proliferation, glucose metabolism, and self-limited T cell persistence; while 4-1BB-based CARs induce a less potent effector T cell but stimulate lipid oxidation and support greater T cell persistence, indicating the potential to differentially reprogram T cells using CARs.

    T Cell Reprogramming by mRNA

    In vitro-transcribed (IVT) mRNA has become an attractive tool in T cell reprogramming and immunotherapeutics because of its rapid and facile production, as well as its safety profile. Unlike DNA, IVT mRNA has no risk of causing insertional mutagenesis and no long-term concern for side effects because of its labile nature.

    Schematic illustration of IVT mRNA use in adoptive T cell therapy. Fig.2 Schematic illustration of IVT mRNA use in adoptive T cell therapy. (Foster, 2019)
    • IVT mRNA for Expanded T Cells and Engineered TCRs
    • IVT mRNA is a useful tool to optimize and test TCR functionality. Mitchell et al. showed that CMV-activated T cells were preferentially transfected with IVT mRNA encoding CXCR2 and those with CXCR2 showed increased migration and trafficking toward CXCR2-specific chemokines for both in vitro and in vivo models.

    • IVT mRNA to Generate CAR T Cells
    • IVT mRNA has been widely used in adoptive T cell therapy, especially in the generation of T cells with CARs. It offers a much safer CAR therapy than viral CARs, since IVT-mRNA leaves no ultimate genetic residue in recipient cells. Foster et al. reported that T cells electroporated with nucleoside-modified and purified mRNA encoding CD19 CAR showed an initial twofold increase in CAR surface expression, as well as a twofold improvement in the cytotoxic killing of leukemia cells.

    Services at Creative Biolabs

    Combined with adequate expertise and advanced instruments in mRNA reprogramming, our scientists have developed a non-integrative IVT mRNA platform to provide T cells reprogramming services expanding beyond TCR and CAR development for immunotherapy. We use mRNA electroporation to reprogram T cells specificity and function to monitor T cell response in immunotherapy.

    At Creative Biolabs, we offer one-stop mRNA-based T cell reprogramming solutions to meet specific application and program needs, expanding from IVT mRNA design, modification, production, to T-cell transfection and characterization. Notably, our technology platform allows in vitro transcription of long RNA strands (~ 10 kbp) with appropriate chemical modification. By investigating a variety of electroporation conditions, we can achieve >90% efficiency and >80% viability when electroporating IVT mRNA into T lymphocytes.

    mRNA-based reprogramming method contributes to the generation of engineered T cells and advances the improvement of efficient T cell reprogramming and T cell immunotherapy. If you are interested in our services, please feel free to contact us.

    Reference

    1. Mitchell, D. A.; et al. Selective modification of antigen-specific T cells by RNA electroporation. Human Gene Therapy. 2008, 19(5): 511-521.
    2. Foster, J. B.; et al. The Emerging Role of In Vitro-Transcribed mRNA in Adoptive T Cell Immunotherapy. Molecular Therapy. 2019, 27(4): 747-756.
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