Delivery of modified mRNA encoding vesicular stomatitis virus matrix protein for colon cancer gene therapy.

Yuquan Wei,Li Yang,Rong Huang,Guonian Zhu,Ke Men,Xueyan Zhang,Rui Zhang,Xingmei Duan,Rongsheng Tong

doi:10.1039/c7ra13656k

Abstract

Plasmid DNA based gene delivery has been widely utilized among both pre-clinical and clinical gene therapy studies. However, therapeutic efficiency is usually limited by the size and potential immune-stimulation issue of plasmid backbone. As an alternative form of genetic material, chemically modified messenger RNA (mRNA) provides a promising alternative to plasmid DNA. In this work, an in vitro transcription mRNA encoding vesicular stomatitis virus matrix protein (VSVMP) was delivered by a cationic liposome–protamine complex, resulting in high mRNA transporting and expression efficiency. The liposome–protamine complex delivered VSVMP mRNA strongly inhibits the growth of C26 tumor cells through inducing apoptosis, while obvious tumor regressions were achieved on both abdominal cavity metastatic and subcutaneous xenograft models in vivo with high safety. Our results also demonstrated that the liposome–protamine–mRNA complex was as potent as its plasmid DNA counterpart, showing strong potential in further colon cancer therapy.

Highlights

Cancer is one of the leading causes of death in both economically developed and developing countries.[1]
vesicular stomatitis virus matrix protein (VSVMP) messenger RNA (mRNA) was synthesized through a T7 polymerase-based in vitro transcription method based on previously constructed VSVMP encoding plasmid pVAX1-VSVMP
Since the length of mRNA might directly affect delivery efficiency and that of Enhanced GFP (EGFP) mRNA used in our experiment was 996 bases long, our results further indicated that VSVMP mRNA (690 bases in total) delivered by CLPP would be highly expressed in cells within 24 hours

Summary

Introduction

Cancer is one of the leading causes of death in both economically developed and developing countries.[1]. In our previous works, delivering apoptosis-inducing genes such as VSVMP and survivin-T34A has been evaluated on several cancer models and desired therapeutic effects were achieved.[8,9,10] the delivery and expression efficiency of the therapeutic gene was highly restricted by the size of plasmids in certain circumstances. This always results in tremendous efforts and costs on optimizing delivery vectors. Developing alternative forms of therapeutic gene is crucial to further facilitate non-viral vector-based therapy

Methods

Results

Conclusion