提高重组型腺相关病毒转导效率的研究现状

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.007

摘要/Abstract

摘要： 重组型腺相关病毒(recombinant adeno-associated virus，rAAV)载体是目前基因治疗研究中常用的、非常有前景的载体之一。欧洲第一个批准上市的基因治疗药物正是基于rAAV。然而，rAAV的转导效率相对有限，导致其治疗成本过高；且过高剂量的rAAV可以激发人体的免疫反应，降低其疗效。因此，如何提高rAAV的转导效率一直是基因治疗领域研究的热点之一。目前常用的提高rAAV转导效率的方法有：使用组织特异性强的血清型/变体、应用蛋白酶体抑制剂、突变衣壳蛋白表面裸露氨基酸、增加单链DNA的第二链合成、构建自身互补型双链载体等。就这些方法各自的原理、应用现状及优劣势进行系统地综述。

关键词: 重组腺相关病毒载体, 基因治疗, 转导效率

Abstract: The recombinant adeno-associated virus(rAAV)vector has emerged as one of the promising and commonly-used vectors in gene therapy research. The first gene therapy drug in clinic approved in Europe is based on rAAV. Due to the relatively limited transduction efficiency, the cost of the rAAV-mediated treatment is expensive. Additionally, high dose of rAAV may trigger host immune response, resulting in the curative effect reduced. Therefore, how to enhance the transduction efficiency of rAAV has been a hot issue in gene therapy. Hitherto there are five common methods to achieve this goal：selecting tissue-specific tropism serotypes and variants, using proteasome inhibitors, mutating capsid surface-exposed amino-acids, increasing second-strand DNA synthesis, and producing self-complementary vectors. In this paper we systemically review the above methods from the aspects of principle, status of application, advantages and disadvantages.

Key words: recombinant adeno-associated virus vector, gene therapy, transduction efficiency

殷子斐, 王丽娜, 王园, 凌晨. 提高重组型腺相关病毒转导效率的研究现状[J]. 生物技术通报, 2015, 31(9): 49-59.

Yin Zifei, Wang Li’na, Wang Yuan, Ling Chen. Research Advances on Increasing the Transduction Efficiency of Recombinant Adeno-associated Viral Vectors[J]. Biotechnology Bulletin, 2015, 31(9): 49-59.

参考文献

[1]Grieger JC, Samulski RJ. Adeno-associated virus vectorology, manufacturing, and clinical applications[J]. Methods Enzymol, 2012, 507：229-254.
[2]Griesenbach U, Alton EW. Current status and future directions of gene and cell therapy for cystic fibrosis[J]. BioDrugs, 2011, 25(2)：77-88.
[3]Mingozzi F, High KA. Therapeutic in vivo gene transfer for genetic disease using AAV：progress and challenges[J]. Nat Rev Genet, 2011, 12(5)：341-355.
[4]Tal J. Adeno-associated virus-based vectors in gene therapy[J]. J Biomed Sci, 2000, 7(4)：279-291.
[5]Masat E, Pavani G, Mingozzi F. Humoral immunity to AAV vectors in gene therapy：challenges and potential solutions[J]. Discov Med, 2013, 15(85)：379-389.
[6]Mingozzi F, High KA. Immune responses to AAV in clinical trials[J]. Curr Gene Ther, 2011, 11(4)：321-330.
[7]Hareendran S, Balakrishnan B, Sen D, et al. Adeno-associated virus(AAV)vectors in gene therapy：immune challenges and strategies to circumvent them[J]. Rev Med Virol, 2013, 23(6)：399-413.
[8]Logan GJ, Alexander IE. Adeno-associated virus vectors：immunobiology and potential use for immune modulation[J]. Curr Gene Ther, 2012, 12(4)：333-343.
[9]刁勇, 许瑞安. 重组腺相关病毒载体诱导的天然免疫反应及机制[J]. 微生物学报, 2012, 52(5)：550-557.
[10]张凤兰, 文朝阳, 丁卫. 腺相关病毒基因治疗载体的改良与应用[J]. 首都医科大学学报, 2009, 30(4)：565-572.
[11]Qing K, Khuntirat B, Mah C, et al. Adeno-associated virus type 2-mediated gene transfer：correlation of tyrosine phosphorylation of the cellular single-stranded D sequence-binding protein with transgene expression in human cells in vitro and murine tissues in vivo[J]. J Virol, 1998, 72(2)：1593-1599.
[12]Zhong L, Li B, Mah CS, et al. Next generation of adeno-associated virus 2 vectors：point mutations in tyrosines lead to high-efficiency transduction at lower doses[J]. Proc Natl Acad Sci USA, 2008, 105(22)：7827-732.
[13]Seisenberger G, Ried MU, Endress T, et al. Real-time single-molecule imaging of the infection pathway of an adeno-associated virus[J]. Science, 2001, 294(5548)：1929-1932.
[14]Nonnenmacher M, Weber T. Adeno-associated virus 2 infection requires endocytosis through the CLIC/GEEC pathway[J]. Cell Host Microbe, 2011, 10(6)：563-576.
[15]Johnson JS, Gentzsch M, Zhang L, et al. AAV exploits subcellular stress associated with inflammation, endoplasmic reticulum expansion, and misfolded proteins in models of cystic fibrosis[J]. PLoS Pathog, 2011, 7(5)：e1002053.
[16]Wu Z, Asokan A, Samulski RJ. Adeno-associated virus serotypes：vector toolkit for human gene therapy[J]. Mol Ther, 2006, 14(3)：316-327.
[17]Gao GP, Alvira MR, Wang L, et al. Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy[J]. Proc Natl Acad Sci USA, 2002, 99(18)：11854-11859.
[18]Miyake K, Miyake N, Yamazaki Y, et al. Serotype-independent method of recombinant adeno-associated virus(AAV)vector production and purification[J]. J Nippon Med Sch, 2012, 79(6)：394-402.
[19]Ling C, Lu Y, Cheng B, et al. High-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors[J]. J Vis Exp, 2011, (49)：doo：10.3791/2538.
[20]Ling C, Lu Y, Kalsi JK, et al. Human hepatocyte growth factor receptor is a cellular coreceptor for adeno-associated virus serotype 3[J]. Hum Gene Ther, 2010, 21(12)：1741-1747.
[21]Cheng B, Ling C, Dai Y, et al. Development of optimized AAV3 serotype vectors：mechanism of high-efficiency transduction of human liver cancer cells[J]. Gene Ther, 2012, 19(4)：375-384.
[22]Markakis EA, Vives KP, Bober J, et al. Comparative transduction efficiency of AAV vector serotypes 1-6 in the substantia nigra and striatum of the primate brain[J]. Mol Ther, 2010, 18(3)：588-593.
[23]Aschauer DF, Kreuz S, Rumpel S. Analysis of transduction efficiency, tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain[J]. PLoS One, 2013, 8(9)：e76310.
[24]Bartel MA, Weinstein JR, Schaffer DV. Directed evolution of novel adeno-associated viruses for therapeutic gene delivery[J]. Gene Ther, 2012, 19(6)：694-700.
[25]Perabo L, Endell J, King S, et al. Combinatorial engineering of a gene therapy vector：directed evolution of adeno-associated virus[J]. J Gene Med, 2006, 8(2)：155-162.
[26]Gray SJ, Blake BL, Criswell HE, et al. Directed evolution of a novel adeno-associated virus(AAV)vector that crosses the seizure-compromised blood-brain barrier(BBB)[J]. Mol Ther, 2010, 18(3)：570-578.
[27]Kienle E, Senis E, B?rner K, et al. Engineering and evolution of synthetic adeno-associated virus(AAV)gene therapy vectors via DNA family shuffling[J]. J Vis Exp, 2012, (62). pii：3819. doi：10.3791/3819
[28]Yang L, Jiang J, Drouin LM, et al. A myocardium tropic adeno-associated virus(AAV)evolved by DNA shuffling and in vivo selection[J]. Proc Natl Acad Sci USA, 2009, 106(10)：3946-3951.
[29]Koerber JT, Schaffer DV. Transposon-based mutagenesis generates diverse adeno-associated viral libraries with novel gene delivery properties[J]. Methods Mol Biol, 2008：434161-434170.
[30]Muller OJ, Kaul F, Weitzman MD, et al. Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors[J]. Nat Biotechnol, 2003, 21(9)：1040-1046.
[31]Varadi K, Michelfelder S, Korff T, et al. Novel random peptide libraries displayed on AAV serotype 9 for selection of endothelial cell-directed gene transfer vectors[J]. Gene Ther, 2012, 19(8)：800-809.
[32]Lisowski L, Dane AP, Chu K, et al. Selection and evaluation of clinically relevant AAV variants in a xenograft liver model[J]. Nature, 2014, 506(7488)：382-386.
[33]Marsic D, Govindasamy L, Currlin S, et al. Vector design tour de force：integrating combinatorial and rational approaches to derive novel adeno-associated virus variants[J]. Mol Ther, 2014, 22(11)：1900-1909.
[34]Engel K, Bassermann F. The ubiquitin proteasome system and its implications for oncology[J]. Dtsch Med Wochenschr, 2013, 138(22)：1178-1182.
[35]Shen M, Schmitt S, Buac D, et al. Targeting the ubiquitin-proteasome system for cancer therapy[J]. Expert Opin Ther Targets, 2013, 17(9)：1091-1108.
[36]Mitchell AM, Samulski RJ. Mechanistic insights into the enhancement of adeno-associated virus transduction by proteasome inhibitors[J]. J Virol, 2013, 87(23)：13035-13041.
[37]Duan D, Yue Y, Yan Z, et al. Endosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus[J]. J Clin Invest, 2000, 105(11)：1573-1587.
[38]Yan Z, Zak R, Luxton GW, et al. Ubiquitination of both adeno-associated virus type 2 and 5 capsid proteins affects the transduction efficiency of recombinant vectors[J]. J Virol, 2002, 76(5)：2043-2053.
[39]Jennings K, Miyamae T, Traister R, et al. Proteasome inhibition enhances AAV-mediated transgene expression in human synoviocytes in vitro and in vivo[J]. Mol Ther, 2005, 11(4)：600-607.
[40]Przystal JM, Umukoro E, Stoneham CA, et al. Proteasome inhibition in cancer is associated with enhanced tumor targeting by the adeno-associated virus/phage[J]. Mol Oncol, 2013, 7(1)：55-66.
[41]Neukirchen J, Meier A, Rohrbeck A, et al. The proteasome inhibitor bortezomib acts differently in combination with p53 gene transfer or cytotoxic chemotherapy on NSCLC cells[J]. Cancer Gene Ther, 2007, 14(4)：431-439.
[42]Monahan PE, Lothrop CD, Sun J, et al. Proteasome inhibitors enhance gene delivery by AAV virus vectors expressing large genomes in hemophilia mouse and dog models：a strategy for broad clinical application[J]. Mol Ther, 2010, 18(11)：1907-1916.
[43]Zhang FL, Jia SQ, Zheng SP, et al. Celastrol enhances AAV1-mediated gene expression in mice adipose tissues[J]. Gene Ther, 2011, 18(2)：128-134.
[44]Wang LN, Wang Y, Lu Y, et al. Pristimerin enhances recombinant adeno-associated virus vector-mediated transgene expression in human cell lines in vitro and murine hepatocytes in vivo[J]. J Integr Med, 2014, 12(1)：20-34.
[45]Ling C, Wang Y, Zhang Y, et al. Selective in vivo targeting of human liver tumors by optimized AAV3 vectors in a murine xenograft model[J]. Hum Gene Ther, 2014, 25(12)：1023-1034.
[46]Rampen A J, Jongen J L, van Heuvel I, et al. Bortezomib-induced polyneuropathy[J]. Neth J Med, 2013, 71(3)：128-133.
[47]Voortman J, Giaccone G. Severe reversible cardiac failure after bortezomib treatment combined with chemotherapy in a non-small cell lung cancer patient：a case report[J]. BMC Cancer, 2006, (6)：129.
[48]Petrucci MT, Giraldo P, Corradini P, et al. A prospective, international phase 2 study of bortezomib retreatment in patients with relapsed multiple myeloma[J]. Br J Haematol, 2013, 160(5)：649-659.
[49]Zhong L, Zhao W, Wu J, et al. A dual role of EGFR protein tyrosine kinase signaling in ubiquitination of AAV2 capsids and viral second-strand DNA synthesis[J]. Mol Ther, 2007, 15(7)：1323-1330.
[50]Zhong L, Li B, Jayandharan G, et al. Tyrosine-phosphorylation of AAV2 vectors and its consequences on viral intracellular trafficking and transgene expression[J]. Virology, 2008, 381(2)：194-202.
[51]Markusic DM, Herzog RW, Aslanidi GV, et al. High-efficiency transduction and correction of murine hemophilia B using AAV2 vectors devoid of multiple surface-exposed tyrosines[J]. Mol Ther, 2010, 18(12)：2048-2056.
[52]Qi YF, Li QH, Shenoy V, et al. Comparison of the transduction efficiency of tyrosine-mutant adeno-associated virus serotype vectors in kidney[J]. Clin Exp Pharmacol Physiol, 2013, 40(1)：53-55.
[53]Qiao C, Zhang W, Yuan Z, et al. Adeno-associated virus serotype 6 capsid tyrosine-to-phenylalanine mutations improve gene transfer to skeletal muscle[J]. Hum Gene Ther, 2010, 21(10)：1343-1348.
[54]Klimczak RR, Koerber JT, Dalkara D, et al. A novel adeno-associated viral variant for efficient and selective intravitreal transduction of rat Muller cells[J]. PLoS One, 2009, 4(10)：e7467.
[55]Dalkara D, Kolstad KD, Guerin KI, et al. AAV mediated GDNF secretion from retinal glia slows down retinal degeneration in a rat model of retinitis pigmentosa[J]. Mol Ther, 2011, 19(9)：1602-1608.
[56]Aslanidi GV, Rivers AE, Ortiz L, et al. High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors[J]. Vaccine, 2012, 30(26)：3908-3917.
[57]Aslanidi GV, Rivers AE, Ortiz L, et al. Optimization of the capsid of recombinant adeno-associated virus 2(AAV2)vectors：the final threshold?[J]. PLoS One, 2013, 8(3)：e59142.
[58]Zolotukhin I, Luo D, Gorbatyuk O, et al. Improved adeno-associated viral gene transfer to murine glioma[J]. J Genet Syndr Gene Ther, 2013, 4(133)：12815.
[59]Dai X, Han J, Qi Y, et al. AAV-mediated lysophosphatidylcholine acyltransferase 1(Lpcat1)gene replacement therapy rescues retinal degeneration in rd11 mice[J]. Invest Ophthalmol Vis Sci, 2014, 55(3)：1724-1734.
[60]Hakim CH, Yue Y, Shin JH, et al. Systemic gene transfer reveals distinctive muscle transduction profile of tyrosine mutant AAV-1, -6, and -9 in neonatal dogs[J]. Mol Ther Methods Clin Dev, 2014, 1：14002.
[61]Mowat FM, Gornik KR, Dinculescu A, et al. Tyrosine capsid-mutant AAV vectors for gene delivery to the canine retina from a subretinal or intravitreal approach[J]. Gene Ther, 2014, 21(1)：96-105.
[62]Qiao C, Yuan Z, Li J, et al. Single tyrosine mutation in AAV8 and AAV9 capsids is insufficient to enhance gene delivery to skeletal muscle and heart[J]. Human Gene Therapy, Part B：Methods, 2012, 23(1)：29-37.
[63]Ferrari FK, Samulski T, Shenk T, et al. Second-strand synthesis is a rate-limiting step for efficient transduction by recombinant adeno-associated virus vectors[J]. J Virol, 1996, 70(5)：3227-3234.
[64]Qing K, Wang XS, Kube DM, et al. Role of tyrosine phosphorylation of a cellular protein in adeno-associated virus 2-mediated transgene expression[J]. Proc Natl Acad Sci USA, 1997, 94(20)：10879-10884.
[65]Mah C, Qing K, Khuntirat B, et al. Adeno-associated virus type 2-mediated gene transfer：role of epidermal growth factor receptor protein tyrosine kinase in transgene expression[J]. J Virol, 1998, 72(12)：9835-9843.
[66]Qing K, Hansen J, Weigel-Kelley KA, et al. Adeno-associated virus type 2-mediated gene transfer：role of cellular FKBP52 protein in transgene expression[J]. J Virol, 2001, 75(19)：8968-8976.
[67] Qing K, Li W, Zhong L, et al. Adeno-associated virus type 2-mediated gene transfer：role of cellular T-cell protein tyrosine phosphatase in transgene expression in established cell lines in vitro and transgenic mice in vivo[J]. J Virol, 2003, 77(4)：2741-2746.
[68]Zhong L, Chen L, Li Y, et al. Self-complementary adeno-associated virus 2(AAV)-T cell protein tyrosine phosphatase vectors as helper viruses to improve transduction efficiency of conventional single-stranded AAV vectors in vitro and in vivo[J]. Mol Ther, 2004, 10(5)：950-957.
[69]Zhao W, Wu J, Zhong L, et al. Adeno-associated virus 2-mediated gene transfer：role of a cellular serine/threonine protein phosphatase in augmenting transduction efficiency[J]. Gene Ther, 2007, 14(6)：545-550.
[70]Jayandharan GR, Zhong L, Li B, et al. Strategies for improving the transduction efficiency of single-stranded adeno-associated virus vectors in vitro and in vivo[J]. Gene Ther, 2008, 15(18)：1287-1293.
[71]Ma W, Li B, Ling C, et al. A simple method to increase the transduction efficiency of single-stranded adeno-associated virus vectors in vitro and in vivo[J]. Hum Gene Ther, 2011, 22(5)：633-640.
[72]McCarty DM, Monahan PE, Samulski RJ. Self-complementary recombinant adeno-associated virus(scAAV)vectors promote efficient transduction independently of DNA synthesis[J]. Gene Ther, 2001, 8(16)：1248-1254.
[73]McCarty DM, Fu H, Monahan PE, et al. Adeno-associated virus terminal repeat(TR)mutant generates self-complementary vectors to overcome the rate-limiting step to transduction in vivo[J]. Gene Ther, 2003, 10(26)：2112-2118.
[74]Wang Z, Ma HI, Li J, et al. Rapid and highly efficient transduction by double-stranded adeno-associated virus vectors in vitro and in vivo[J]. Gene Ther, 2003, 10(26)：2105-2111.
[75]Nathwani AC, Gray JT, Ng CY, et al. Self-complementary adeno-associated virus vectors containing a novel liver-specific human factor IX expression cassette enable highly efficient transduction of murine and nonhuman primate liver[J]. Blood, 2006, 107(7)：2653-2661.
[76]Gao GP, Lu Y, Sun X, et al. High-level transgene expression in nonhuman primate liver with novel adeno-associated virus serotypes containing self-complementary genomes[J]. J Virol, 2006, 80(12)：6192-6194.
[77]Liu Y, Keefe K, Tang X, et al. Use of self-complementary adeno-associated virus serotype 2 as a tracer for labeling axons：implications for axon regeneration[J]. PLoS One, 2014, 9(2)：e87447.
[78]McCarty DM. Self-complementary AAV vectors；advances and applications[J]. Mol Ther, 2008, 16(10)：1648-1656.
[79]Ding W, Yan Z, Zak R, et al. Second-strand genome conversion of adeno-associated virus type 2(AAV-2)and AAV-5 is not rate limiting following apical infection of polarized human airway epithelia[J]. J Virol, 2003, 77(13)：7361-7366.
[80]Martino AT, Suzuki M, Markusic DM, et al. The genome of self-complementary adeno-associated viral vectors increases Toll-like receptor 9-dependent innate immune responses in the liver[J]. Blood, 2011, 117(24)：6459-6468.
[81]Wu T, Topfer K, Lin S W, et al. Self-complementary AAVs induce more potent transgene product-specific immune responses compar-ed to a single-stranded genome[J]. Mol Ther, 2012, 20(3)：572-579.
[82]Mitchell AM, Li C, Samulski RJ. Arsenic trioxide stabilizes accumulations of adeno-associated virus virions at the perinuclear region, increasing transduction in vitro and in vivo[J]. J Virol, 2013, 87(8)：4571-4583.
[83]Le HT, Yu QC, Wilson JM, et al. Utility of PEGylated recombinant adeno-associated viruses for gene transfer[J]. J Control Release, 2005, 108(1)：161-177.
[84]Ponnazhagan S, Mahendra G, Kumar S, et al. Conjugate-based targeting of recombinant adeno-associated virus type 2 vectors by using avidin-linked ligands[J]. J Virol, 2002, 76(24)：12900-12907.
[85]Nathwani AC, Tuddenham EG, Rangarajan S, et al. Adenovirus-associated virus vector-mediated gene transfer in hemophilia B[J]. N Engl J Med, 2011, 365(25)：2357-2365.