T Cell Engineering Using mRNA
Adoptive T cell therapy is a promising form of cellular therapy. With continuous development over the past several decades, T cell-based immunotherapy has made periodic progress with US Food and Drug Administration (FDA) approval of chimeric antigen receptor (CAR) T cell therapy for leukemia and lymphoma. Adoptive T cell transfer generally requires ex vivo modification of isolated autologous cells. Therefore, a number of compounds and techniques for T cell engineering have been developed and evaluated, including in vitro-transcribed (IVT) mRNA. Compared to the conventional use of viral vectors, IVT mRNA has numerous benefits, including inherent safety features, efficient recombinant protein translation, as well as the ability to control pharmacokinetic properties of the therapy. Creative Biogene is a forward-looking research institute as well as a leading custom service provider in the field of mRNA-based cellular engineering and reprogramming. We are committed to achieving the most prominent modification of T cells, including the induction of cancer-specific T cell receptors CAR and TCR expression. Our services are supported by both experienced experts and the latest advanced technologies. We not only offer electroporation method but also have the capability to design ionizable lipid nanoparticles (LNPs) for ex vivo mRNA delivery to human T cells.
mRNA transfection opens the door to the generation of T cell engineering
Adoptive T cell therapy uses human immune cells, usually by expressing recombinant proteins, to attack selected targets on tumors or infected cells. The current standard for manufacturing genetic engineering of T cells is mainly based on the use of retro- or lentiviral transduction. However, the method has a risk of insertional mutagenesis. The use of mRNA transfection for modification of T cells is a promising strategy to mitigate the adverse effects associated with viral vectors. It allows directed integration of T cell receptors to the corresponding locus, leading to uniform T cell receptor expression as well as enhancing T cell potency. In addition, mRNA encoding CARs or TCRs can avoid the occurrence of cytokine release syndrome due to its temporary expression of CARs or TCRs.
Advantage of mRNA transfection for T cell engineering compared to the conventional use of viral vectors.
- Reduces the potential for viral contamination.
- Less time- and resource-intensive.
- Transient expression of the T cell receptors.
- Reduce the risk for off-target effects.
mRNA-based T cell engineering in Creative Biogene
In general, the use of mRNA transfection for T cell engineering includes isolation and expanding T cells, transfection of IVT mRNA, and cell quality control. Among them, transfection of IVT mRNA is a crucial step as well as cell culture. In addition to optimizing T cell isolation and expansion methods, we also offer two strategies for achieving mRNA transfection. We offer electroporation of mRNA and LNPs-mediated mRNA delivery to facilitate the successful GMP-compliant production of CAR-T cells.
mRNA-Based T cell engineering.
Electroporation for T cell mRNA delivery
Electroporation has been used to deliver a broader range of bioactive constructs into a variety of cell types. T cells are a type of sensitive cells and can be difficult to transfect. Based on our advanced modular electroporator, we can achieve T cell mRNA transfection in a flexible and scalable way. With years of accumulated experience in electroporation conditions for T cells, we can support the delivery of IVT mRNA into T cells with transfection rates of >80 % and survival rates of >80 %.
Ionizable lipid nanoparticle (LNPs)- mediated mRNA delivery for modification of T cells
Electroporation is the most common method for human T cell mRNA delivery, which bypasses the extensive safety and regulatory requirements for T cell engineering using viruses. However, electroporation's negative impact on cell viability and function limit this physical transfection method for more demanding T cell engineering. Therefore, we offer LNPs for ex vivo mRNA delivery to human T cells. After formulation and screening of ionizable lipids, we can offer the efficient and top-performing LNP formulation for CAR or TCR mRNA delivery to human T cells.
If you are interested in our services, please don't hesitate to contact us. We look forward to providing services for your next project.
- Foster, J. B., et al. (2019). "The emerging role of in vitro-transcribed mRNA in adoptive T cell immunotherapy." Molecular Therapy, 27(4), 747-756.