基因治疗用腺相关病毒载体的剂量效应与剂量控制

摘要/Abstract

摘要： 人类基因治疗需要在靶细胞适时、适量、安全的产生具有疗效的药物,这些正好是定量药理学的研究范畴。重组腺相关病毒载体(rAAV)具有的各种特性以及近来在规模化制备方面取得的进展使其逐渐成为基因治疗的理想载体。超过60项以AAV为载体的基因治疗已经完成或正在展开。其中,rAAV载体的剂量与基因表达效率以及临床疗效之间的关系是研究者关注的焦点之一。多数研究认为,在一定的剂量范围内,基因表达水平具有剂量依赖性。然而, 基因表达水平同时还受病毒载体构型、组织趋向性、细胞靶向性、注射途径, 甚至患者的年龄、性别等条件影响 ,基因表达水平还可受到其他药物的调控。因此,基因治疗实验过程中应结合定量药理学深入展开,解决基因治疗中的定量与药效之间的关系。

关键词: 基因治疗, 病毒载体, 剂量效应, 定量药理学, 临床治疗

Abstract: Human gene therapy needs to express exogenous DNA at the targeting cells, producing a practical and efficient therapeutic dosage at an approp-riate time (quantitative pharmacology)with a safe man-ner .Recombinant adeno-associated virus(rAAV)vec-tors possess a number of properties and recent progress in rAAV production made it rapidly become the reagent of choice for therapeutic gene transfer.Over 60 clinical trials of gene therapy based on rAAV have been carried out .The dose response reaction between rAAV vectors and gene expression activity or clinical outcome is one of major aspects of these trials.Most studies showed that vector genomes (vg)and gene expression had a concentration-dependent relationship during a certain scope .However, gene expression can be affected by viral serotypes, tissue tropisms, cell targeting , drug regulation, injection route , age and sex , etc .Thus, these aspects should be carefully considered by scienti-sts, pharmacologists and physicians during animal ex-periments or clinical trails.

Key words: gene therapy, viral vector, dose-re-sponse, quantitative pharmacology, clinical therapy

中图分类号:

R968

王启钊, 吕颖慧, 许瑞安. 基因治疗用腺相关病毒载体的剂量效应与剂量控制[J]. 中国临床药理学与治疗学, 2008, 13(10): 1182-1194.

WANG Qi-zhao, LV Ying-hui, XU Rui-an. Dose-response and control of adeno-associated viral vectors based preclinical and clinical gene therapy[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(10): 1182-1194.

参考文献

[1] Minchin RF, Orr RJ, Cronin AS, et al.The pharmacolo-gy of gene therapy[J] .CMJ, 1999, 40(3):381-391.
[2] Xiao X, Li J, Samulski RJ.Production of high-titer recombinant adeno-associated virus vectors in the absence of helper adenovirus[J] .J Virol, 1998, 72(3):2224-2232.
[3] Stender S, Murphy M, O'BrienT, et al. Adeno-associat-ed viral vector transduction of human mesenchymal stem cells[J] .Eur CellMater, 2007, 13:93-99.
[4] Nakai H, Thomas CE, Storm TA, et al.A Limited num-ber of transducible hepatocytes restrictsa wide-range linear vector dose response in recombinant adeno-associated vir-us-mediated liver transduction[J] .J Virol, 2002, 6(22): 11343-11349.
[5] Forsayeth JR, Eberling JL, Sanftner LM, et al.A dose-ranging study of AAV-hAADC therapy in Parkinsonian monkeys[J] .Mol Ther, 2006, 14(4):571-757.
[6] Jacobson SG, Acland GM, Aguirre GD, et al.Safety of recombinant adeno-associated virus type 2-RPE65 vector delivered by ocular subretinal injection[J] .Mol Ther, 2006, 13(6):1074-1084.
[7] Cao L, Liu Y, During MJ, et al.High-titer, wild-type free recombinant adeno-associated virus vector production using intron-containing helper plasmids[J] .J Virol, 2000, 74(24):11456-11463.
[8] Ogura T, Mizukami H, Mimuro J, et al.Utility of intrap-eritoneal administration as a route of AAV serotype 5 vec-tor-mediated neonatal gene transfer[J] .J Gene Med, 2006, 8(8):990-997.
[9] Kaplitt MG, Feigin A, Tang C, et al.Safety and tolera-bility of gene therapy with an adeno-associated virus (AAV)borne GAD gene for Parkinson' s disease:An open-label, phase I trial[J] .Lancet, 2007, 369(9579): 2097-2105.
[10] Xu RA, Janson CG, Mastakov M, et al.Quantitative comparison of expression with adeno-associated virus (AAV-2)brain-specific gene cassettes[J] .Gene Ther, 2001, 8(17):1323-1332.
[11] Liu Y, Okada T, Nomoto T, et al.Promoter effectsof ad-eno-associated viral vector for transgene expression in the cochlea in vivo[J] .Exp Mol Med, 2007, 39(2):170-175.
[12] Halbert CL, Lam SL, Miller AD.High-efficiency promot-er-dependent transduction by adeno-associated virus type 6 vectors in mouse lung[J] .Hum Gene Ther, 2007, 18(4):344-354.
[13] Gossen M, Bujard H.Tight control of gene expression in mammalian cells by tetracycline-responsive[J] .Proc Natl Acad Sci USA, 1992, 89(12):5547-5551.
[14] Chtarto A, Yang X, Bockstael O, et al.Controlled deliv-ery of glial cell line-derivedneurotrophic factor by a single tetracycline-inducible AAV vector[J] .Exp Neurol, 2007, 204(1):387-399.
[15] Stieger K, Le Meur G, Lasne F, et al.Long-term doxycy-cline-regulated transgene expression in the retina of non-human primates following subretinal injection of recombi-nant AAV vectors[J] .Mol Ther, 2006, 13(5):967-975.
[16] Wang J, VoutetakisA, Papa M, et al. Rapamycin control of transgene expression from a single AAV vector in mouse salivary glands[J] .Gene Ther, 2006, 13(2):187-190.
[17] Su H, Kan YW.Adeno-associated viral vector-delivered hypoxia-inducible gene expression in ischemic hearts[J] . MethodsMol Biol, 2007, 366:331-342.
[18] Ogasawara Y, Mizukami H, Urabe M, et al.Highly regu-lated expression of adeno-associated virus large Rep pro-teins in stable 293 cell lines using the Cre loxP switching system[J] .J Gen Virol, 1999, 80(9):2477-2480.
[19] Wu Z, Asokan A, Samulski RJ.Adeno-associated virus serotypes:vector tookit for human gene therapy[J] .Mol Ther, 2006, 14(3):316-327.
[20] Limberis MP, Wilson JM.Adeno-associated virus serotype 9 vectors transduce murine alveolar and nasal epithelia and can be readministered[J] .Proc Natl Acad Sci USA, 2006, 103(35):12993-12998.
[21] Bostick B, Ghosh A, Yue Y, et al.Systemic AAV-9 transduction in mice is influenced by animal age but not by the route of administration[J] .Gene Ther, 2007, 14(22):1605-1609.
[22] Broekman ML, Comer LA, Hyman BT, et al.Adeno-as-sociated virus vectors serotyped with AAV8 capsid are more efficient than AAV-1 or -2 serotypes for widespread gene delivery to the neonatal mouse brain[J] .Neuro-science, 2006, 138(2):501-510.
[23] Harding TC, Dickinson PJ, Roberts BN, et al.Enhanced gene transfer efficiency in the murine striatum and an or-thotopic glioblastoma tumor model, using AAV-7-and AAV-8-pseudotyped vectors[J] .Hum Gene Ther, 2006, 17(8):807-820.
[24] Palomeque J, Chemaly ER, Colosi P, et al.Efficiency of eight different AAV serotypes in transducing rat myocardi-um in vivo[J] .Gene Ther, 2007, 14(13):989-997.
[25] Pacak CA, MahCS, Thattaliyath BD, et al. Recombinant adeno-associated virus serotype 9 leads to preferential car-diac transduction in vivo[J] .Circ Res, 2006, 99(4):e3-9.
[26] Allocca M, Mussolino C, Garcia-Hoyos M, et al.Novel adeno-associated virus serotypes efficiently transduce mu-rine photoreceptors[J] .J Virol, 2007, 81(20):11372-11380.
[27] Takeda S, Takahashi M, Mizukami H, et al.Successful gene transfer using adeno-associated virus vectors into the kidney:comparison among adeno-associated virus serotype 1-5 vectors in vitro and in vivo[J] .Nephron Exp Neph-rol, 2004, 96(4):e119-126.
[28] Wang AY, Peng PD, Ehrhardt A, et al.Comparison of adenoviral and adeno-associated viral vectorsfor pancreat-ic gene delivery in vivo[J] .Hum Gene Ther, 2004, 15(4):405-413.
[29] Cheng H, Wolfe SH, Valencia V, et al.Efficient and persistent transduction of exocrine and endocrine pancreas by adeno-associated virus type 8[J] .J Biomed Sci, 2007, 14(5):585-594.
[30] Rivière C, Danos O, Douar AM.Long-term expression and repeated administration of AAV type 1, 2 and 5 vec-tors in skeletal muscle of immunocompetent adult mice[J] .Gene Ther, 2006, 13(17):1300-1308.
[31] Ghosh A, Yue Y, Duan D.Viral serotype and the trans-gene sequence influence overlapping adeno-associated vi-ral (AAV)vector-mediated gene transfer in skeletal mus-cle[J] .J Gene Med, 2006, 8(3):298-305.
[32] Taymans JM, Vandenberghe LH, Haute CV, et al.Com-parative analysis of adeno-associated viral vector serotypes 1, 2, 5, 7, and 8 inmouse brain[J] .Hum Gene Ther, 2007, 18(3):195-206.
[33] Lux K, Goerlitz N, Schlemminger S, et al.Green fluores-cent protein-tagged adeno-associated virus particles allow the study of cytosolic and nuclear trafficking[J] .J Virol, 2005, 79(18):11776-11787.
[34] Shi J, Zheng D, Liu Y, et al.Over-expression of soluble TRAIL induces apoptosis in human lung adenocarcinoma and inhibits growth of tumor xenografts in nude mice[J] . Cancer Res, 2005, 65(5):1687-1689.
[35] Fleurence E, Riviere C, Lacaze-Masmonteil T, et al.Comparative efficacy of intratracheal adeno-associated vir-us administration to newborn rats[J] .Hum Gene Ther, 2005, 16(11):1298-1306.
[36] Cearley CN, Wolfe JH.Transduction characteristicsof ad-eno-associated virus vectors expressing cap serotypes 7, 8, 9, and Rh10in the mouse brain[J] .MolTher, 2006, 13(3):528-537.
[37] Ledley TS, Ledley FD.Multicompartment, numerical model of cellular events in the pharmacokinetics of gene therapies[J] .Hum Gene Ther, 1994, 5(6):679-691.
[38] Qing K, Mah C, Hansen J, et al.Human fibroblast growth factor receptor 1 is a co-receptor for infection by adeno-associated virus 2[J] .Nat Med, 1999, 5(1):71-77.
[39] Summerford C, Bartlett JS, Samulski RJ.AlphaVbeta5 in-tegrin:a co-receptor for adeno-associated virus type 2 in-fection[J] .Nat Med, 1999, 5(1):78-82.
[40] Bartlett JS, Wilcher R, Samulski RJ.Infectious entry pathway of adeno-associated virus and adeno-associated virus vectors[J] .J Virol, 2000, 74(6):2777-2785.
[41] Xu RA, Herrison P, Chen M, et al.Cytoglobin overex-pression protects against damage-induced fibrosis[J] .Mol Ther, 2006, 13(6):1093-1000.
[42] Duan D, Li Q, Kao AW, et al.Dynamin is required for recombinant adeno-associated virus type 2 infection[J] .J Virol, 1999, 73(12):10371-10376.
[43] Douar AM, Poulard K, Stockholm D, et al.Intracellular trafficking of adeno-associatedvirus vectors:routing to the late endosomal compartment and proteasome degradation[J] .J Virol, 2001, 75(4):1824-1833.
[44] Bantel -Schaal U, Hub B, Kartenbeck J.Endocytosis of adeno-associated virus type 5 leads to accumulationof vir-us particles in the Golgi compartment[J] .J Virol, 2002, 76(5):2340-2349.
[45] Ding W, Zhang L N, Yeaman C, et al.rAAV2 traffics through both the late and the recycling endosomes in a dose-dependent fashion[J] .Mol Ther, 2006, 13(4):671-682.
[46] Xiao W, WarringtonKH Jr, Hearing P, et al. Adenovir-us-facilitated nuclear translocation of adeno-associated vir-us type 2[J] .J Virol, 2002, 76(22):11505-11517.
[47] Yokoi K, Kachi S, Zhang HS, et al.Ocular gene transfer with self -complementary AAV vectors[J] .Invest Ophthal-mol Vis Sci, 2007, 48(7):3324-3328.
[48] Choi VW, McCarty DM, Samulski RJ.Host cell DNA re-pair pathways in adeno-associated viral genome processing[J] .J Virol, 2006, 80(21):10346-10356.
[49] Streck CJ, Dickson PV, Ng CY, et al.Adeno-associated virus vector-mediated systemic delivery of IFN-beta com-bined with low-dose cyclophosphamide affects tumor re-gression in murine neuroblastoma models[J] .Clin Cancer Res, 2005, 11(16):6020-6029.
[50] Cotugno G, Pollock R, Formisano P, et al.Pharmacologi-cal regulation of the insulin receptor signaling pathway mimics insulin action in cells transducedwith viralvectors[J] .Hum Gene Ther, 2004, 15(11):1101-1108.
[51] Wang Z, Zhu T, Rehman KK, et al.Widespreadand sta-ble pancreatic gene transfer by adeno-associated virus vec-tors via different routes[J] .Diabetes, 2006b, 55(4):875-884.
[52] Conrad CK, Allen SS, Afione SA, et al.Safety of single-dose administration of an adeno-associated virus(AAV)-CFTR vector in the primate lung[J] .Gene Ther, 1996, 3(8):658-668.
[53] Lai L, Davision BB, Veazey RS, et al.A preliminary evaluation of recombinant adeno-assiociated virus biodis-tribution in rhesus monkeys after intrahepatic inoculation inutero[J] .Hum Gene Ther, 2002, 13(17):2027-2039.
[54] Favre D, Provost N, Blouin V, et al.Immediate and long-term safety of recombinant adeno-associated virus in-jection into the nonhuman primate muscle[J] .Mol ther, 2001, 4(6):559-566.
[55] Watson G, Bastacky J, Belichenko P, et al.Intrathecal administration of AAV vectors for the treatment of lysoso-mal storage in the brains of MPS I mice[J] .Gene Ther, 2006, 13(11):917-925.
[56] Ito T, Okada T, Mimuro J, et al.Adenoassociated virus-mediated prostacyclin synthase expression prevents pulmo-nary arterial hypertension in rats[J] .Hypertension, 2007, 50(3):531-536.
[57] Cearley CN, Wolfe JH.A single injection of an adeno-as-sociated virus vector into nuclei with divergent connections results in widespread vector distribution in the brain and global correction of a neurogenetic disease[J] .J Neuros-ci, 2007, 27(37):9928-9940.
[58] Di Pasquale G, Chiorini JA.AAV transcytosis through barrier epithelia and endothelium[J] .MolTher, 2006, 13(3):506-516.
[59] Rodino-Klapac LR, Janssen PML, Montgomery CL, et al.A translational approach for limb vascular delivery of the micro-dystrophin gene without high volume or high pres-sure for treatment of Duchenne muscular dystrophy[J] .J Transl Med, 2007, 5(45):1-11.
[60] Mori S, Takeuchi T, Enomoto Y, et al.Biodistribution of a low dose of intravenously administered AAV -2, 10, and 11 vectors to cynomolgus monkeys[J] .Jpn J Infect Dis, 2006, 59(5):285-293.
[61] Mah C, Cresawn KO, FraitesTJ, et al. Sustained correc-tion of glycogen storage disease type II using adeno-associ-atedvirus serotype 1 vectors[J] .Gene Ther, 2005, 12: 1405-1409.
[62] Ding Z, Georgiev P, Thöny B.Administration-route and gender-independent long-term therapeutic correction of phenylketonuria (PKU)in a mouse model by recombinant adeno-associated virus 8 pseudotyped vector-mediated gene transfer[J] .Gene Ther, 2006, 13(7):587-593.
[63] Passini MA, Bu J, Fidler JA, et al.Combination brain and systemic injectionsof AAV provide maximal functional and survival benefits in the Niemann-Pick mouse[J] . Proc NatlAcad Sci USA, 2007, 104(22):9505-9510.
[64] During M, Xu R, Young D, et al.Peroral gene therapy of lactose intolerance using an adeno-associated virus vector[J] .Nat Med, 1998, 4(10):1131-1135.
[65] During MJ, Symes CW, Lawlor PA, et al.An oral vac-cine against NMDAR1with efficacy in experimental stroke and epilepsy[J] .Science, 2000, 287(5457):1453-1460.
[66] Ma H, Liu Y, Liu S, et al.Oral adeno-associated virus-sTRAIL gene therapy suppresses human hepatocellular carcinoma growth inmice[J] .Hepatology, 2005, 42(6): 1355-1363.
[67] Hara H, Inoue M, Adachi K, et al.Mucosal immunother-apy for alzheimer' s disease with viral vectors[J] .Nihon Shinkei Seishin Yakurigaku Zasshi, 2007, 27(2):53-56.
[68] Mouri A, Noda Y, Hara H, et al.Oralvaccination with a viral vector containing Abeta cDNA attenuates age-related Abeta accumulation and memory deficits without causing inflammation in a mouse Alzheimer model[J] .FASEB J, 2007, 21(9):2135-2148.
[69] Sumner-Jones SG, Gill DR, Hyde SC.Lack of repeat transduction by rAAV5 5 vectors in the mouse airway[J] . J Virol, 2007, 81(22):12360-12367.