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Enhanced Delivery and Potency of Self-Amplifying mRNA Vaccines by Electroporation in Situ.

Cu Y, Broderick KE, Banerjee K, Hickman J, Otten G, Barnett S, Kichaev G, Sardesai NY, Ulmer JB, Geall A - Vaccines (Basel) (2013)

Bottom Line: Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications.In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein.These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

View Article: PubMed Central - PubMed

Affiliation: Novartis Vaccines & Diagnostics, Inc., 350 Massachusetts Ave, Cambridge, MA 02139, USA.

ABSTRACT
Nucleic acid-based vaccines such as viral vectors, plasmid DNA (pDNA), and mRNA are being developed as a means to address limitations of both live-attenuated and subunit vaccines. DNA vaccines have been shown to be potent in a wide variety of animal species and several products are now licensed for commercial veterinary but not human use. Electroporation delivery technologies have been shown to improve the generation of T and B cell responses from synthetic DNA vaccines in many animal species and now in humans. However, parallel RNA approaches have lagged due to potential issues of potency and production. Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications. In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein. These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

No MeSH data available.


In vivo expression of secreted embryonic alkaline phosphatase (SEAP) after intramuscular njection of self-amplifying mRNA (RNA, 1 µg or 10 µg, panels A,C) or pDNA (DNA, 1 µg or 10 µg, panels B,D). The vectors were delivered by bilateral intramuscular injection with (+EP) or without electroporation. Serum SEAP expression was measured on days 1, 3, 10 and 17 after treatment, and represented as the log of mean relative luminescence (RLU)±SEM, n = 5/group. * Statistically significant (student-t test, p < 0.05).
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vaccines-01-00367-f001: In vivo expression of secreted embryonic alkaline phosphatase (SEAP) after intramuscular njection of self-amplifying mRNA (RNA, 1 µg or 10 µg, panels A,C) or pDNA (DNA, 1 µg or 10 µg, panels B,D). The vectors were delivered by bilateral intramuscular injection with (+EP) or without electroporation. Serum SEAP expression was measured on days 1, 3, 10 and 17 after treatment, and represented as the log of mean relative luminescence (RLU)±SEM, n = 5/group. * Statistically significant (student-t test, p < 0.05).

Mentions: After bilateral intramuscular injection of a low dose (1 μg) of naked self-amplifying mRNA encoding for the transgene secreted alkaline phosphatase (SEAP), measureable but low levels of serum SEAP were detectable as early as 3 days after treatment. No significant enhancements in the levels of SEAP expression were observed when the RNA was delivered using EP (RNA + EP) at the same dose, except at the latest time point (Figure 1A). In contrast to the RNA, mice injected with a low dose (1 μg) of naked pDNA showed levels of SEAP expression that were indistinguishable from baseline (Figure 1B), but substantial enhancements in SEAP expression was observed when pDNA was delivered by EP (pDNA + EP) at the same dose. At the 1 μg dose, SEAP expression by pDNA + EP was superior to RNA + EP.


Enhanced Delivery and Potency of Self-Amplifying mRNA Vaccines by Electroporation in Situ.

Cu Y, Broderick KE, Banerjee K, Hickman J, Otten G, Barnett S, Kichaev G, Sardesai NY, Ulmer JB, Geall A - Vaccines (Basel) (2013)

In vivo expression of secreted embryonic alkaline phosphatase (SEAP) after intramuscular njection of self-amplifying mRNA (RNA, 1 µg or 10 µg, panels A,C) or pDNA (DNA, 1 µg or 10 µg, panels B,D). The vectors were delivered by bilateral intramuscular injection with (+EP) or without electroporation. Serum SEAP expression was measured on days 1, 3, 10 and 17 after treatment, and represented as the log of mean relative luminescence (RLU)±SEM, n = 5/group. * Statistically significant (student-t test, p < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4494232&req=5

vaccines-01-00367-f001: In vivo expression of secreted embryonic alkaline phosphatase (SEAP) after intramuscular njection of self-amplifying mRNA (RNA, 1 µg or 10 µg, panels A,C) or pDNA (DNA, 1 µg or 10 µg, panels B,D). The vectors were delivered by bilateral intramuscular injection with (+EP) or without electroporation. Serum SEAP expression was measured on days 1, 3, 10 and 17 after treatment, and represented as the log of mean relative luminescence (RLU)±SEM, n = 5/group. * Statistically significant (student-t test, p < 0.05).
Mentions: After bilateral intramuscular injection of a low dose (1 μg) of naked self-amplifying mRNA encoding for the transgene secreted alkaline phosphatase (SEAP), measureable but low levels of serum SEAP were detectable as early as 3 days after treatment. No significant enhancements in the levels of SEAP expression were observed when the RNA was delivered using EP (RNA + EP) at the same dose, except at the latest time point (Figure 1A). In contrast to the RNA, mice injected with a low dose (1 μg) of naked pDNA showed levels of SEAP expression that were indistinguishable from baseline (Figure 1B), but substantial enhancements in SEAP expression was observed when pDNA was delivered by EP (pDNA + EP) at the same dose. At the 1 μg dose, SEAP expression by pDNA + EP was superior to RNA + EP.

Bottom Line: Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications.In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein.These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

View Article: PubMed Central - PubMed

Affiliation: Novartis Vaccines & Diagnostics, Inc., 350 Massachusetts Ave, Cambridge, MA 02139, USA.

ABSTRACT
Nucleic acid-based vaccines such as viral vectors, plasmid DNA (pDNA), and mRNA are being developed as a means to address limitations of both live-attenuated and subunit vaccines. DNA vaccines have been shown to be potent in a wide variety of animal species and several products are now licensed for commercial veterinary but not human use. Electroporation delivery technologies have been shown to improve the generation of T and B cell responses from synthetic DNA vaccines in many animal species and now in humans. However, parallel RNA approaches have lagged due to potential issues of potency and production. Many of the obstacles to mRNA vaccine development have recently been addressed, resulting in a revival in the use of non-amplifying and self-amplifying mRNA for vaccine and gene therapy applications. In this paper, we explore the utility of EP for the in vivo delivery of large, self-amplifying mRNA, as measured by reporter gene expression and immunogenicity of genes encoding HIV envelope protein. These studies demonstrated that EP delivery of self-amplifying mRNA elicited strong and broad immune responses in mice, which were comparable to those induced by EP delivery of pDNA.

No MeSH data available.