modRNA技术:一种蛋白表达新方法
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摘要
早在上世纪90年代,注射裸信使核糖核酸(mRNA)已经能够在体内实现外源性蛋白的表达。然而,mRNA的不稳定性及高度免疫原性阻碍了这项技术的进一步发展与应用。近年来,随着对mRNA结构的认识深入及核酸技术的进步,可化学合成并修饰信使核糖核酸,即Chemically synthetic modified message RNA(modRNA),modRNA兼具稳定性、低免疫原性及良好的蛋白翻译表的优势。我们对其研究现状、存在问题进行综述,并对可能的改进方向进行展望。
Earlier in the 1990s, direct injection of naked mRNA has been proved to be an approach to achieve the exogenous gene expression in vivo. However, the instability and high-immunogenicity of m RNA becomes the main challenges of further development and application of this technology. Recently, with the in-depth research of m RNA structure and the advancement of nucleic acid technologies, chemically synthetic modified message RNA(modRNA) has emerged as a newlydeveloped gene vector, featuring stability, low-immunogenicity and high expression of target proteins. Based on the related researches, the concept, structure, mechanism and research status of modRNA were concluded, and the advantages and flaws along with the potential improvement direction were put forward in this paper.
Earlier in the 1990s, direct injection of naked mRNA has been proved to be an approach to achieve the exogenous gene expression in vivo. However, the instability and high-immunogenicity of m RNA becomes the main challenges of further development and application of this technology. Recently, with the in-depth research of m RNA structure and the advancement of nucleic acid technologies, chemically synthetic modified message RNA(modRNA) has emerged as a newlydeveloped gene vector, featuring stability, low-immunogenicity and high expression of target proteins. Based on the related researches, the concept, structure, mechanism and research status of modRNA were concluded, and the advantages and flaws along with the potential improvement direction were put forward in this paper.
引文
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[27]Thess A,Grund S,Mui BL,et al.Sequence-engineered mRNAwithout chemical nucleoside modifications enables an effective protein therapy in large animals[J].Mol Ther,2015,23(9):1456-1464.
[28]Pardi N,Hogan MJ,Pelc RS,et al.Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination[J].Nature,2017,543(7644):248-251.
[29]Richner JM,Himansu S,Dowd KA,et al.Modified mRNAvaccines protect against zika virus infection[J].Cell,2017,168(6):1114-1125.
[30]Sahin U,Derhovanessian E,Miller M,et al.Personalized RNAmutanome vaccines mobilize poly-specific therapeutic immunity against cancer[J].Nature,2017,547(7662):222-226.
[31]Stadler CR,Bahr-Mahmud H,Celik L,et al.Elimination of large tumors in mice by mRNA-encoded bispecific antibodies[J].Nat Med,2017,23(7):815-817.
[32]Islam MA,Reesor EK,Xu Y,et al.Biomaterials for mRNA delivery[J].Biomater Sci,2015,3(12):1519-1533.
[2]Alexopoulou L,Holt AC,Medzhitov R,et al.Recognition of doublestranded RNA and activation of NF-kappaB by Toll-like receptor 3[J].Nature,2001,413(6857):732-738.
[3]Heil F,Hemmi H,Hochrein H,et al.Species-specific recognition of single-stranded RNA via toll-like receptor 7 and 8[J].Science,2004,303(5663):1526-1529.
[4]Hornung V,Ellegast J,Kim S,et al.5'-triphosphate RNA is the ligand for rig-i[J].Science,2006,314(5801):994-997.
[5]Pichlmair A,Schulz O,Tan CP,et al.Activation of mda5 requires higher-order RNA structures generated during virus infection[J].J Virol,2009,83(20):10761-10769.
[6]Sahin U,Kariko K,Tureci O.mRNA-based therapeutics-developing a new class of drugs[J].Nat Rev Drug Discov,2014,13(10):759-780.
[7]KarikóK,Buckstein M,Ni H,et al.Suppression of RNA recognition by toll-like receptors:The impact of nucleoside modification and the evolutionary origin of RNA[J].Immunity,2005,23(2):165-175.
[8]Anderson BR,Muramatsu H,Nallagatla SR,et al.Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation[J].Nucleic Acids Res,2010,38(17):5884-5892.
[9]Anderson BR,Muramatsu H,Jha BK,et al.Nucleoside modifications in RNA limit activation of 2'-5'-oligoadenylate synthetase and increase resistance to cleavage by RNAse l[J].Nucleic Acids Res,2011,39(21):9329-9338.
[10]Kariko K,Muramat su H,Welsh FA,et al.Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector with increased t ranslational capacity and biological stability[J].Mol Ther,2008,16(11):1833-1840.
[11]Andries O,Mc Cafferty S,De Smedt SC,et al.N(1)-methylpseudouridine-incorporated mRNA outperforms pseudouridineincorporated mRNA by providing enhanced protein expression and reduced immunogenicity in mammalian cell lines and mice[J].J Control Release,2015,217:337-344.
[12]Hulot JS,Ishikawa K,Hajjar RJ.Gene therapy for the treatment of heart failure:Promise postponed[J].Eur Heart J,2016,37(21):1651-1658.
[13]Warren L,Manos PD,Ahfeldt T,et al.Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA[J].Cell Stem Cell,2010,7(5):618-630.
[14]Luni C,Giulitti S,Serena E,et al.High-efficiency cellular reprogramming with microfluidics[J].Nat Methods,2016,13(5):446-452.
[15]Carlsson L,Clarke JC,Yen C,et al.Biocompatible,purified vegf-a mRNA improves cardiac function after intracardiac injection1 week post-myocardial infarction in swine[J].Mol Ther Methods Clin Dev,2018,9:330-346.
[16]Lui KO,Zangi L,Silva EA,et al.Driving vascular endothelial cell fate of human multipotent Isl1+heart progenitors with VEGFmodified mRNA[J].Cell Res,2013,23(10):1172-1186.
[17]Zangi L,Lui KO,von Gise A,et al.Modified mRNA directs the fate of heart progenitor cells and induces vascular regeneration after myocardial infarction[J].Nat Biotechnol,2013,31(10):898-907.
[18]Huang CL,Leblond AL,Turner EC,et al.Synthetic chemically modified mRNA-based delivery of cytoprotective factor promotes early cardiomyocyte survival post-acute myocardial infarction[J].Mol Pharm,2015,12(3):991-996.
[19]Zangi L,Oliveira MS,Ye LY,et al.Insulin-like growth factor 1receptor-dependent pathway drives epicardial adipose tissue formation after myocardial injury[J].Circulation,2017,135(1):59-72.
[20]Chien KR,Zangi L,Lui KO.Synthetic chemically modified mRNA(modRNA):Toward a new technology platform for cardiovascular biology and medicine[J].Cold Spring Harb Perspect Med,2014,5(1):a014035.
[21]Balmayor ER,Geiger JP,Aneja MK,et al.Chemically modified RNA induces osteogenesis of stem cells and human tissue explants as well as accelerates bone healing in rats[J].Biomaterials,2016,87:131-146.
[22]Elangovan S,Khorsand B,Do AV,et al.Chemically modified RNA activated matrices enhance bone regeneration[J].J Control Release,2015,218:22-28.
[23]Khorsand B,Elangovan S,Hong L,et al.A comparative study of the bone regenerative effect of chemically modified RNA encoding BMP-2 or BMP-9[J].AAPS J,2017,19(2):438-446.
[24]Wang G,McCain ML,Yang L,et al.Modeling the mitochondrial cardiomyopathy of barth syndrome with induced pluripotent stem cell and heart-on-chip technologies[J].Nat Med,2014,20(6):616-623.
[25]Kormann MS,Hasenpusch G,Aneja MK,et al.Expression of therapeutic proteins after delivery of chemically modified mRNAin mice[J].Nat Biotechnol,2011,29(2):154-157.
[26]Kariko K,Muramatsu H,Keller JM,et al.Increased erythropoiesis in mice injected with submicrogram quantities of pseudouridinecontaining mRNA encoding erythropoietin[J].Mol Ther,2012,20(5):948-953.
[27]Thess A,Grund S,Mui BL,et al.Sequence-engineered mRNAwithout chemical nucleoside modifications enables an effective protein therapy in large animals[J].Mol Ther,2015,23(9):1456-1464.
[28]Pardi N,Hogan MJ,Pelc RS,et al.Zika virus protection by a single low-dose nucleoside-modified mRNA vaccination[J].Nature,2017,543(7644):248-251.
[29]Richner JM,Himansu S,Dowd KA,et al.Modified mRNAvaccines protect against zika virus infection[J].Cell,2017,168(6):1114-1125.
[30]Sahin U,Derhovanessian E,Miller M,et al.Personalized RNAmutanome vaccines mobilize poly-specific therapeutic immunity against cancer[J].Nature,2017,547(7662):222-226.
[31]Stadler CR,Bahr-Mahmud H,Celik L,et al.Elimination of large tumors in mice by mRNA-encoded bispecific antibodies[J].Nat Med,2017,23(7):815-817.
[32]Islam MA,Reesor EK,Xu Y,et al.Biomaterials for mRNA delivery[J].Biomater Sci,2015,3(12):1519-1533.