Current Status of Messenger RNA Delivery Systems

Stanton, Matthew

doi:10.1089/nat.2018.0726

Cited by 21 publications

(16 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The other important factor determining an efficient gene transfection is the type of carrier. Various non-viral gene delivery vehicles made of cationic polymers or lipids with different modifications have been developed 27,43 . However, beyond the transfection of established cell lines, only few of them achieved robust gene delivery in primary cells 22,44 .…”

Section: Discussionmentioning

confidence: 99%

mRNA Transfection-Induced Activation of Primary Human Monocytes and Macrophages: Dependence on Carrier System and Nucleotide Modification

Moradian

Roch

Lendlein

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Monocytes and macrophages are key players in maintaining immune homeostasis. identifying strategies to manipulate their functions via gene delivery is thus of great interest for immunological research and biomedical applications. We set out to establish conditions for mRnA transfection in hard-to-transfect primary human monocytes and monocyte-derived macrophages due to the great potential of gene expression from in vitro transcribed mRnA for modulating cell phenotypes. mRnA doses, nucleotide modifications, and different carriers were systematically explored in order to optimize high mRNA transfer rates while minimizing cell stress and immune activation. We selected three commercially available mRnA transfection reagents including liposome and polymer-based formulations, covering different application spectra. Our results demonstrate that liposomal reagents can particularly combine high gene transfer rates with only moderate immune cell activation. For the latter, use of specific nucleotide modifications proved essential. In addition to improving efficacy of gene transfer, our findings address discrete aspects of innate immune activation using cytokine and surface marker expression, as well as cell viability as key readouts to judge overall transfection efficiency. The impact of this study goes beyond optimizing transfection conditions for immune cells, by providing a framework for assessing new gene carrier systems for monocyte and macrophage, tailored to specific applications. Innate immune cells play an important role in response to pathological conditions and maintaining immune homeostasis 1. Among them, monocytes and monocyte-derived macrophages have remarkable properties, including their immunomodulatory capacities. Monocytes with various distinct phenotypes are key players of early inflammation. During inflammation, monocytes can dynamically repolarize to different phenotypes in response to local signals, which is thought to be more efficient at resolving tissue homeostasis than the recruitment of other anti-inflammatory and pro-regenerative subsets of monocytes or macrophages 2-4. Macrophages themselves have special functions such as phagocytosis of invading pathogens or apoptotic cells, antigen presentation to T cells, elimination of pathogens via releasing reactive oxygen species or proteolytic enzymes, and secretion of pro-or anti-inflammatory signaling molecules to recruit various types of other immune cells 5-9. Elucidation and manipulation of monocyte and macrophage phenotypes is therefore essential to fully explore their role in immunoregulation. This will benefit not only basic immunological research but also clinical and translational studies 3,10,11. For innate immune cell manipulation, transfections have been commonly used to introduce nucleic acids, such as plasmid DNA (pDNA) or small interfering RNA, to induce or inhibit the expression of a target protein,

show abstract

Section: Discussionmentioning

confidence: 99%

mRNA Transfection-Induced Activation of Primary Human Monocytes and Macrophages: Dependence on Carrier System and Nucleotide Modification

Moradian

Roch

Lendlein

et al. 2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Several groups are also attempting to create particles that behave like viruses able to shield mRNA from endo-, 5 0 exo-, and 3 0 exo-nucleases, deliver mRNA efficiently into cells, and achieve a sufficient level of the encoded protein. [127][128][129] In all cases, the vehicle must pass through the target cell membrane, and after cell uptake, usually by endocytosis, it must escape the endosome and unload its mRNA cargo into the cytosol, where translation occurs. For some mRNAs, the delivery system may also influence the quantity and quality of local gene expression patterns, innate immune stimulation, and can provide a synergistic adjuvant effect.…”

Section: Self-amplifying Mrna Vaccinesmentioning

confidence: 99%

mRNA as a Transformative Technology for Vaccine Development to Control Infectious Diseases

et al. 2019

View full text Add to dashboard Cite

“…With recent technological advances in oligonucleotide delivery, mRNA therapy in living animals has become a feasible strategy to treat a variety of pathologies (1). All mRNAs are polyanions that vary only in sequence and length, whereas the proteins that they encode can elicit countless functions.…”

Section: Introductionmentioning

confidence: 99%

Local Delivery ofOx40l,Cd80, andCd86mRNA Kindles Global Anticancer Immunity

et al. 2019

View full text Add to dashboard Cite

Localized expression of effector molecules can initiate antitumor responses through engagement of specific receptors on target cells in the tumor microenvironment. These locally induced responses may also have a systemic effect, clearing additional tumors throughout the body. In this study, to evoke systemic antitumor responses, we utilized charge-altering releasable transporters (CART) for local intratumoral delivery of mRNA coding for costimulatory and immune-modulating factors. Intratumoral injection of the CART-mRNA complexes resulted in mRNA expression at the site of administration, transfecting a substantial proportion of tumor-infiltrating dendritic cells, macrophages, and T cells in addition to the tumor cells, resulting in a local antitumor effect. Using a twotumor model, we further show that mRNA therapy locally administered to one tumor stimulated a systemic antitumor response, curing both tumors. The combination of Ox40l-, Cd80-, and Cd86-encoding mRNA resulted in the local upregulation of proinflammatory cytokines, robust local T-cell activation, and migration of immune cells to local draining lymph node or to an anatomically distant tumor. This approach delayed tumor growth, facilitated tumor regression, and cured tumors in both A20 and CT26 tumor models. These results highlight mRNA-CART therapy as a viable approach to induce systemic antitumor immunity from a single localized injection.Significance: The mRNA-CART system is a highly effective delivery platform for delivering immunostimulatory genes into the tumor microenvironment for potential therapeutic development.

show abstract

Current Status of Messenger RNA Delivery Systems

Cited by 21 publications

References 53 publications

mRNA Transfection-Induced Activation of Primary Human Monocytes and Macrophages: Dependence on Carrier System and Nucleotide Modification

mRNA Transfection-Induced Activation of Primary Human Monocytes and Macrophages: Dependence on Carrier System and Nucleotide Modification

mRNA as a Transformative Technology for Vaccine Development to Control Infectious Diseases

Local Delivery ofOx40l,Cd80, andCd86mRNA Kindles Global Anticancer Immunity

Contact Info

Product

Resources

About