Design and Development of Modified mRNA Encoding Core Antigen of Hepatitis C Virus: a Possible Application in Vaccine Production

Abstract

Background: Hepatitis C virus (HCV) is a blood-borne pathogen, resulting in liver cirrhosis and liver cancer. Despite of many efforts in development of treatments for HCV, no vaccine has been licensed yet. The purpose of this study was to design and prepare a specific mRNA, without 5' cap and poly (A) tail transcribed in vitro capable of coding core protein and also to determine its functionality. Methods: Candidate mRNA was prepared by in vitro transcription of the designed construct consisting of 5ʹ and 3ʹ untranslated regions of heat shock proteins 70 (hsp70) mRNA, T7 promoter, internal ribosome entry site (IRES) sequences of eIF4G related to human dendritic cells (DCs), and the Core gene of HCV. To design the modified mRNA, the 5' cap and poly (A) tail structures were not considered. DCs were transfected by in vitro-transcribed messenger RNA (IVT-mRNA) and the expressions of green fluorescent protein (GFP), and Core genes were determined by microscopic examination and Western blotting assay. Results: Cell transfection results showed that despite the absence of 5' cap and poly (A) tail, the structure of the mRNA was stable. Moreover, the successful expressions of GFP and Core genes were achieved. Conclusion: Our findings indicated the effectiveness of a designed IVT-mRNA harboring the Core gene of HCV in transfecting and expressing the antigens in DCs. Considering the simple and efficient protocol for the preparation of this IVT-mRNA and its effectiveness in expressing the gene that it carries, this IVT-mRNA could be suitable for development of an RNA vaccine against HCV.

Hepatitis C virus DNA vaccines: a systematic review

“…The preliminary reports on mass vaccination with SARS-CoV-2 mRNA vaccines have demonstrated their general safety and high efficacy in disease prevention [ 71 ]. Therefore, with proper backbone, mRNAs encoding HCV antigens [ 72 ] could obviate the need for electroporation to generate sufficient immune response and demonstrate higher efficacy and immunogenicity compared to DNA vaccines.…”

Nanoplatforms for mRNA Therapeutics

“…[ 137 ] Formulations based on PLGA have been reported to deliver various types of cargos, such as hydrophobic drugs, proteins, and nucleic acid. [ 137 ] In addition to polyplexes, [ 138 ] PLGA has also been incorporated into LPPs for mRNA delivery. [ 139 ] mRNA delivery efficiency from different polymer‐based LPPs has been investigated and compared.…”

In the era of rapid mRNA-based vaccines: Why is there no effective hepatitis C virus vaccine yet?

“…On the contrary, with the exception of using mRNA to transfect DCs (which will be discussed in the next section), there have been no pre-clinical or clinical trials using mRNA-based vaccines against HCV. Interestingly, Sharifnia et al [ 282 ] (2019) have proposed for the first time that an RNA-based vaccine against HCV could be feasible since after in vitro generation of an mRNA coding for the core protein, they were able to detect core protein in monocyte-derived DCs which were previously transfected with this construct[ 282 ]. Unfortunately, no further animal studies were performed to assess the immunogenicity of this approach.…”