Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo

被引:185
作者
Kuhn, A. N. [1 ]
Diken, M. [1 ]
Kreiter, S. [1 ]
Selmi, A. [1 ]
Kowalska, J. [2 ]
Jemielity, J. [2 ]
Darzynkiewicz, E. [2 ]
Huber, C. [1 ]
Tureci, O. [1 ]
Sahin, U. [1 ]
机构
[1] Johannes Gutenberg Univ Mainz, Div Translat & Expt Oncol, Dept Internal Med 3, D-55131 Mainz, Germany
[2] Univ Warsaw, Inst Expt Phys, Fac Phys, Div Biophys, Warsaw, Poland
关键词
vaccination; dendritic cells; RNA; cap analog; TRANSFECTED DENDRITIC CELLS; MESSENGER-RNA; VACCINATION TRIAL; DIRECT-INJECTION; GENE-THERAPY; CANCER; EXPRESSION; PROTEIN; VITRO; DEGRADATION;
D O I
10.1038/gt.2010.52
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Vaccination with in vitro transcribed RNA coding for tumor antigens is considered a promising approach for cancer immunotherapy and has already entered human clinical testing. One of the basic objectives for development of RNA as a drug is the optimization of immunobioavailability of the encoded antigen in vivo. By analyzing the effect of different synthetic 5' mRNA cap analogs on the kinetics of the encoded protein, we found that m(2)(7,2)' (O)Gpp(s)pG (beta-S-ARCA) phosphorothioate caps, in particular the D1 diastereoisomer, profoundly enhance RNA stability and translational efficiency in immature but not mature dendritic cells. Moreover, in vivo delivery of the antigen as beta-S-ARCA(D1)-capped RNA species is superior for protein expression and for efficient priming and expansion of naive antigen-specific T cells in mice. Our findings establish 5 0 mRNA cap analogs as yet another module for tuning immunopharmacological properties of recombinant antigen-encoding RNA for vaccination purposes. Gene Therapy (2010) 17, 961-971; doi: 10.1038/gt.2010.52; published online 22 April 2010
引用
收藏
页码:961 / 971
页数:11
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