A synthetic DNA template for fast manufacturing of versatile single epitope mRNA

Wout de Mey; Phaedra De Schrijver; Dorien Autaers; Lena Pfitzer; Bruno Fant; Hanne Locy; Arthur Esprit; Lien Lybaert; Cedric Bogaert; Magali Verdonck; Kris Thielemans; Karine Breckpot; Lorenzo Franceschini

doi:10.1016/j.omtn.2022.08.021

A synthetic DNA template for fast manufacturing of versatile single epitope mRNA

Wout de Mey, Phaedra De Schrijver, Dorien Autaers, Lena Pfitzer, Bruno Fant, Hanne Locy, Arthur Esprit, Lien Lybaert, Cedric Bogaert, Magali Verdonck, Kris Thielemans, Karine Breckpot, Lorenzo Franceschini

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Abstract

A flexible, affordable, and rapid vaccine platform is necessary to unlock the potential of personalized cancer vaccines in order to achieve full clinical efficiency. mRNA cancer vaccine manufacture relies on the rigid sequence design of multiepitope constructs produced by laborious bacterial cloning and time-consuming plasmid preparation. Here, we introduce a synthetic DNA template (SDT) assembly process, which allows cost- and time-efficient manufacturing of single (neo)epitope mRNA. We benchmarked SDT-derived mRNA against mRNA derived from a plasmid DNA template (PDT), showing that monocyte-derived dendritic cells (moDCs) electroporated with SDT-mRNA or PDT-mRNA, encoding HLA-I- or HLA-II-restricted (neo)epitopes, equally activated T cells that were modified to express the cognate T cell receptors. Furthermore, we validated the SDT-mRNA platform for neoepitope immunogenicity screening using the characterized HLA-A2-restricted neoepitope DHX40B and four new candidate HLA-A2-restricted melanoma neoepitopes. Finally, we compared SDT-mRNA with PDT-mRNA for vaccine development purposes. moDCs electroporated with mRNA encoding the HLA-A2-restricted, mutated Melan-A/Mart-1 epitope together with TriMix mRNA-generated high levels of functional Melan-A/Mart-1-specific CD8+ T cells. In conclusion, SDT single epitope mRNA can be manufactured in a more flexible, cost-efficient, and time-efficient way compared with PDT-mRNA, allowing prompt neoepitope immunogenicity screening, and might be exploited for the development of personalized cancer vaccines.

Original language	English
Pages (from-to)	943-954
Number of pages	12
Journal	Molecular Therapy Nucleic Acids
Volume	29
DOIs	https://doi.org/10.1016/j.omtn.2022.08.021
Publication status	Published - 13 Sept 2022

Bibliographical note

Keywords

MT: Oligonucleotides: Therapies and Applications
T cell
cancer
mRNA
neoantigen
synthetic DNA template

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10.1016/j.omtn.2022.08.021Licence: CC BY

88346845Final published version, 1.28 MBLicence: CC BY

Cite this

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title = "A synthetic DNA template for fast manufacturing of versatile single epitope mRNA",

abstract = "A flexible, affordable, and rapid vaccine platform is necessary to unlock the potential of personalized cancer vaccines in order to achieve full clinical efficiency. mRNA cancer vaccine manufacture relies on the rigid sequence design of multiepitope constructs produced by laborious bacterial cloning and time-consuming plasmid preparation. Here, we introduce a synthetic DNA template (SDT) assembly process, which allows cost- and time-efficient manufacturing of single (neo)epitope mRNA. We benchmarked SDT-derived mRNA against mRNA derived from a plasmid DNA template (PDT), showing that monocyte-derived dendritic cells (moDCs) electroporated with SDT-mRNA or PDT-mRNA, encoding HLA-I- or HLA-II-restricted (neo)epitopes, equally activated T cells that were modified to express the cognate T cell receptors. Furthermore, we validated the SDT-mRNA platform for neoepitope immunogenicity screening using the characterized HLA-A2-restricted neoepitope DHX40B and four new candidate HLA-A2-restricted melanoma neoepitopes. Finally, we compared SDT-mRNA with PDT-mRNA for vaccine development purposes. moDCs electroporated with mRNA encoding the HLA-A2-restricted, mutated Melan-A/Mart-1 epitope together with TriMix mRNA-generated high levels of functional Melan-A/Mart-1-specific CD8+ T cells. In conclusion, SDT single epitope mRNA can be manufactured in a more flexible, cost-efficient, and time-efficient way compared with PDT-mRNA, allowing prompt neoepitope immunogenicity screening, and might be exploited for the development of personalized cancer vaccines.",

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day = "13",

doi = "10.1016/j.omtn.2022.08.021",

language = "English",

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journal = "Molecular Therapy Nucleic Acids",

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TY - JOUR

T1 - A synthetic DNA template for fast manufacturing of versatile single epitope mRNA

AU - de Mey, Wout

AU - De Schrijver, Phaedra

AU - Autaers, Dorien

AU - Pfitzer, Lena

AU - Fant, Bruno

AU - Locy, Hanne

AU - Esprit, Arthur

AU - Lybaert, Lien

AU - Bogaert, Cedric

AU - Verdonck, Magali

AU - Thielemans, Kris

AU - Breckpot, Karine

AU - Franceschini, Lorenzo

PY - 2022/9/13

Y1 - 2022/9/13

N2 - A flexible, affordable, and rapid vaccine platform is necessary to unlock the potential of personalized cancer vaccines in order to achieve full clinical efficiency. mRNA cancer vaccine manufacture relies on the rigid sequence design of multiepitope constructs produced by laborious bacterial cloning and time-consuming plasmid preparation. Here, we introduce a synthetic DNA template (SDT) assembly process, which allows cost- and time-efficient manufacturing of single (neo)epitope mRNA. We benchmarked SDT-derived mRNA against mRNA derived from a plasmid DNA template (PDT), showing that monocyte-derived dendritic cells (moDCs) electroporated with SDT-mRNA or PDT-mRNA, encoding HLA-I- or HLA-II-restricted (neo)epitopes, equally activated T cells that were modified to express the cognate T cell receptors. Furthermore, we validated the SDT-mRNA platform for neoepitope immunogenicity screening using the characterized HLA-A2-restricted neoepitope DHX40B and four new candidate HLA-A2-restricted melanoma neoepitopes. Finally, we compared SDT-mRNA with PDT-mRNA for vaccine development purposes. moDCs electroporated with mRNA encoding the HLA-A2-restricted, mutated Melan-A/Mart-1 epitope together with TriMix mRNA-generated high levels of functional Melan-A/Mart-1-specific CD8+ T cells. In conclusion, SDT single epitope mRNA can be manufactured in a more flexible, cost-efficient, and time-efficient way compared with PDT-mRNA, allowing prompt neoepitope immunogenicity screening, and might be exploited for the development of personalized cancer vaccines.

AB - A flexible, affordable, and rapid vaccine platform is necessary to unlock the potential of personalized cancer vaccines in order to achieve full clinical efficiency. mRNA cancer vaccine manufacture relies on the rigid sequence design of multiepitope constructs produced by laborious bacterial cloning and time-consuming plasmid preparation. Here, we introduce a synthetic DNA template (SDT) assembly process, which allows cost- and time-efficient manufacturing of single (neo)epitope mRNA. We benchmarked SDT-derived mRNA against mRNA derived from a plasmid DNA template (PDT), showing that monocyte-derived dendritic cells (moDCs) electroporated with SDT-mRNA or PDT-mRNA, encoding HLA-I- or HLA-II-restricted (neo)epitopes, equally activated T cells that were modified to express the cognate T cell receptors. Furthermore, we validated the SDT-mRNA platform for neoepitope immunogenicity screening using the characterized HLA-A2-restricted neoepitope DHX40B and four new candidate HLA-A2-restricted melanoma neoepitopes. Finally, we compared SDT-mRNA with PDT-mRNA for vaccine development purposes. moDCs electroporated with mRNA encoding the HLA-A2-restricted, mutated Melan-A/Mart-1 epitope together with TriMix mRNA-generated high levels of functional Melan-A/Mart-1-specific CD8+ T cells. In conclusion, SDT single epitope mRNA can be manufactured in a more flexible, cost-efficient, and time-efficient way compared with PDT-mRNA, allowing prompt neoepitope immunogenicity screening, and might be exploited for the development of personalized cancer vaccines.

KW - MT: Oligonucleotides: Therapies and Applications

KW - T cell

KW - cancer

KW - mRNA

KW - neoantigen

KW - synthetic DNA template

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U2 - 10.1016/j.omtn.2022.08.021

DO - 10.1016/j.omtn.2022.08.021

M3 - Article

C2 - 36159589

SN - 2162-2531

VL - 29

SP - 943

EP - 954

JO - Molecular Therapy Nucleic Acids

JF - Molecular Therapy Nucleic Acids

ER -

A synthetic DNA template for fast manufacturing of versatile single epitope mRNA

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

SRP84: SRP-Onderzoekszwaartepunt: PACT: multi-omics Profiling of T cells to improve Adoptive Cell Therapy

Cite this

A synthetic DNA template for fast manufacturing of versatile single epitope mRNA

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Fingerprint

Projects

SRP84: SRP-Onderzoekszwaartepunt: PACT: multi-omics Profiling of T cells to improve Adoptive Cell Therapy

Cite this