mRNA, a Revolution in Biomedicine

Baptista, Bruno; Carapito, Rita; Laroui, Nabila; Pichon, Chantal; Sousa, Fani

doi:10.3390/pharmaceutics13122090

Cited by 31 publications

(25 citation statements)

References 163 publications

(405 reference statements)

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“…In vitro transcribed mRNA can be used as a therapeutic agent in preventive vaccines, cancer immunotherapy, or gene replacement therapy (30)(31)(32). However, the utility of IVT RNA exceeds potential clinical applications.…”

Section: Discussionmentioning

confidence: 99%

2’-O-methylation of the second transcribed nucleotide within mRNA 5’ cap impacts protein production level in a cell specific manner and contributes to RNA immune evasion

Drążkowska¹,

Baran

Warmiński

et al. 2022

Preprint

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In higher eukaryotes, m7G-adjacent nucleotides undergo extensive modifications. Ribose of the first or first and second transcribed nucleotides can be subjected to 2'-O-methylation to form cap1 or cap2, respectively. Additionally, when the first transcribed nucleotide is adenosine, it can not only undergo 2'-O-methylation but can also be methylated at position N6 forming N6,2'-O-dimethyladenosine (m6Am). Recent studies have shed some light on the functions of cap1, showing that cap1 in mammalian cells plays a crucial role in distinguishing between 'self' and 'non-self' RNA during viral infection. Here, we attempted to understand the impact of other cap methylations on RNA-related processes. Therefore, we synthesized tetranucleotide cap analogs and used them for efficient co-transcriptional RNA capping during in vitro transcription. Using this tool, we found that 2'-O-methylation of the second transcribed nucleotide within the mRNA 5' cap influences protein production levels in a cell-specific manner. The presence of this modification can strongly hamper protein biosynthesis or do not influence protein production levels. Interestingly, 2'-O-methylation of the second transcribed nucleotide as well as the presence of N6,2'-O-dimethyladenosine as the first transcribed nucleotide serve as determinants that define transcripts as 'self' and contribute to transcript escape from the host innate immune response. Additionally, cap methylation status does not influence transcript affinity towards translation initiation factor 4E or in vitro susceptibility to decapping by DCP2; however what we observe is resistance of RNA capped with cap2 to DXO-mediated decapping and degradation.

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Section: Discussionmentioning

confidence: 99%

2’-O-methylation of the second transcribed nucleotide within mRNA 5’ cap impacts protein production level in a cell specific manner and contributes to RNA immune evasion

Drążkowska¹,

Baran

Warmiński

et al. 2022

Preprint

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“…Discovered in the 1970s, the 5′-cap structure (m7G5′ppp5′N), composed of a 7-methylguanosine nucleoside and a terminal nucleotide linked through a triphosphate bridge in the 5′-mRNA, confers IVT mRNA stability and translation efficiency [ 42 ]. There are three major types of 5′-cap structures, including type O, type I, and type II, which are classified according to whether there is a methylated ribose on the 2′ hydroxyl group of the first or second nucleotide from the 5′ end [ 43 ] ( Figure 4 ).…”

Section: The Principles Of Mrna Vaccine Design and Modificationmentioning

confidence: 99%

“…The 5′-UTR is a noncoding region, but it can help mRNA bind to ribosomes [ 42 ]. Early research suggested that the secondary structure of the 5′-UTR can inhibit mRNA translation, whose symbol is a high GC content [ 49 , 50 ].…”

Section: The Principles Of Mrna Vaccine Design and Modificationmentioning

confidence: 99%

Nonviral Delivery Systems of mRNA Vaccines for Cancer Gene Therapy

et al. 2022

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In recent years, the use of messenger RNA (mRNA) in the fields of gene therapy, immunotherapy, and stem cell biomedicine has received extensive attention. With the development of scientific technology, mRNA applications for tumor treatment have matured. Since the SARS-CoV-2 infection outbreak in 2019, the development of engineered mRNA and mRNA vaccines has accelerated rapidly. mRNA is easy to produce, scalable, modifiable, and not integrated into the host genome, showing tremendous potential for cancer gene therapy and immunotherapy when used in combination with traditional strategies. The core mechanism of mRNA therapy is vehicle-based delivery of in vitro transcribed mRNA (IVT mRNA), which is large, negatively charged, and easily degradable, into the cytoplasm and subsequent expression of the corresponding proteins. However, effectively delivering mRNA into cells and successfully activating the immune response are the keys to the clinical transformation of mRNA therapy. In this review, we focus on nonviral nanodelivery systems of mRNA vaccines used for cancer gene therapy and immunotherapy.

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“…As a result of these efforts, co-formulation into lipid nanoparticles (LNPs) has become one of the most successful and commonly used methods for mRNA delivery in vivo (Ickenstein LM, et al, 2019). Because of its reliability, mRNA-LNPs are being extensively utilized in a broad range of new and potential treatments, such as regenerative medicines, vaccines, immunotherapies, and gene editing applications (Baptista B, et al, 2021).…”

Section: Fortunately the Hybridoma Technology Developed By Georges Ko...mentioning

confidence: 99%

An Efficient Approach for SARS-CoV-2 Monoclonal Antibody Production via Modified mRNA-LNP Immunization

Hsu

Liang

Kumari

et al. 2022

Preprint

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Throughout the COVID-19 pandemic, many prophylactic and therapeutic drugs have been evaluated and introduced. Among these treatments, monoclonal antibodies (mAbs) that bind to and neutralize SARS-CoV-2 virus have been applied as complementary and alternative treatments to vaccines. Although different methodologies have been utilized to produce mAbs, traditional hybridoma fusion technology is still commonly used for this purpose due to its unmatched performance record. In this study, we coupled the hybridoma fusion strategy with mRNA-lipid nanoparticle (LNP) immunization. This time-saving approach can circumvent biological and technical hurdles, such as difficult to express membrane proteins, antigen instability, and the lack of posttranslational modifications on recombinant antigens. We used mRNA-LNP immunization and hybridoma fusion technology to generate mAbs against the receptor binding domain (RBD) of SARS-CoV-2 spike (S) protein. Compared with traditional protein-based immunization approaches, inoculation of mice with RBD mRNA-LNP induced higher titers of serum antibodies. In addition, the mAbs we obtained can bind to SARS-CoV-2 RBDs from several variants. Notably, RBD-mAb-3 displayed particularly high binding affinities and neutralizing potencies against both Alpha and Delta variants. In addition to introducing specific mAbs against SARS-CoV-2, our data generally demonstrate that mRNA-LNP immunization may be useful to quickly generate highly functional mAbs against emerging infectious diseases.

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mRNA, a Revolution in Biomedicine

Cited by 31 publications

References 163 publications

2’-O-methylation of the second transcribed nucleotide within mRNA 5’ cap impacts protein production level in a cell specific manner and contributes to RNA immune evasion

2’-O-methylation of the second transcribed nucleotide within mRNA 5’ cap impacts protein production level in a cell specific manner and contributes to RNA immune evasion

Nonviral Delivery Systems of mRNA Vaccines for Cancer Gene Therapy

An Efficient Approach for SARS-CoV-2 Monoclonal Antibody Production via Modified mRNA-LNP Immunization

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