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  • ISSN 1006-3080
  • CN 31-1691/TQ

新型肌肉高效亲和AAV血清型的开发

陈颖 吴佳梅 凌菲香 曾步兵 郑静 吴侠 赵锴 肖啸

陈颖, 吴佳梅, 凌菲香, 曾步兵, 郑静, 吴侠, 赵锴, 肖啸. 新型肌肉高效亲和AAV血清型的开发[J]. 华东理工大学学报(自然科学版), 2022, 48(1): 51-58. doi: 10.14135/j.cnki.1006-3080.20210127001
引用本文: 陈颖, 吴佳梅, 凌菲香, 曾步兵, 郑静, 吴侠, 赵锴, 肖啸. 新型肌肉高效亲和AAV血清型的开发[J]. 华东理工大学学报(自然科学版), 2022, 48(1): 51-58. doi: 10.14135/j.cnki.1006-3080.20210127001
CHEN Ying, WU Jiamei, LING Feixiang, ZENG Bubing, ZHENG Jing, WU Xia, ZHAO Kai, XIAO Xiao. Generation of a Novel Muscle-Tropic AAV Serotype[J]. Journal of East China University of Science and Technology, 2022, 48(1): 51-58. doi: 10.14135/j.cnki.1006-3080.20210127001
Citation: CHEN Ying, WU Jiamei, LING Feixiang, ZENG Bubing, ZHENG Jing, WU Xia, ZHAO Kai, XIAO Xiao. Generation of a Novel Muscle-Tropic AAV Serotype[J]. Journal of East China University of Science and Technology, 2022, 48(1): 51-58. doi: 10.14135/j.cnki.1006-3080.20210127001

新型肌肉高效亲和AAV血清型的开发

doi: 10.14135/j.cnki.1006-3080.20210127001
基金项目: 中央高校基本科研业务费专项资金赞助
详细信息
    作者简介:

    陈颖:陈 颖(1995-),女,江苏南通人,硕士生,研究方向为基因治疗的新药研发。E-mail:cysj950925@163.com

    通讯作者:

    肖 啸,E-mail:xiaoxiao@ecust.edu.cn

  • 中图分类号: Q782

Generation of a Novel Muscle-Tropic AAV Serotype

  • 摘要: 通过在AAV5衣壳的N573后插入一个寡肽PGPSPAD生成了一种改良的AAV血清型,称为AAVc1,它在体外和体内均表现出比AAV5、AAV8和AAV9更好的肌肉感染性;恒河猴血清中针对AAVc1的中和抗体(Neutralizing antibodies,Nabs)滴度低于AAV9,这表明针对AAVc1的中和抗体的免疫应答低于AAV9。结果表明,新型血清型AAVc1应用到AAV基因治疗中优于野生型AAV血清型。

     

  • 图  1  从C2C12成肌细胞中筛选AAV5突变衣壳

    Figure  1.  Screen of AAV5 mutant capsids from C2C12 myoblast cell

    图  2  AAV在C2C12成肌细胞中的转导效率

    Figure  2.  Transduction efficiency of AAV in C2C12 myoblast cell

    图  3  AAV在小鼠肌肉中的转导效率

    Figure  3.  Transduction efficiency of AAV in mouse muscle

    表  1  AAV血清型效价的定量

    Table  1.   Quantification of AAV serotype titer

    SerotypeTiter/(vg·mL−1)
    AAV5AAV8AAV9AAVc1
    qPCR1.09×10121.54×10128.95×10111.28×1012
    Silver stain1.65×10123.25×10122.31×10121.93×1012
    下载: 导出CSV

    表  2  猴子血清中不同AAV血清型的中和抗体检测

    Table  2.   Detection of neutralizing antibodies of different AAV serotypes in monkey serum

    Monkey IDDilution ratio
    Anti-AAV9Anti-AAVc1
    115591∶321∶2
    130611∶81∶1
    130731∶161∶2
    131751∶161∶8
    133431∶2<1∶1
    136531∶81∶2
    137251∶321∶8
    800431∶11∶1
    1000391∶161∶1
    12347A
    1∶1<1∶1
    下载: 导出CSV
  • [1] LI C, SAMULSKI R J. Engineering adeno-associated virus vectors for gene therapy[J]. Nature Reviews Genetics, 2020, 21(4): 255-272. doi: 10.1038/s41576-019-0205-4
    [2] NALDINI L. Ex vivo gene transfer and correction for cell-based therapies[J]. Nature Reviews Genetics, 2011, 12(5): 301-315. doi: 10.1038/nrg2985
    [3] MINGOZZI F, HIGH K A. Therapeutic in vivo gene transfer for genetic disease using AAV: Progress and challenges[J]. Nature Reviews Genetics, 2011, 12(5): 341-355. doi: 10.1038/nrg2988
    [4] ATCHISON R W, CASTO B C, HAMMON W M. Adenovirus-associated defective virus particles[J]. Science, 1965, 149(3685): 754-756. doi: 10.1126/science.149.3685.754
    [5] HIRSCH M L, WOLF S J, SAMULSKI R J. Delivering transgenic DNA exceeding the carrying capacity of AAV vectors[J]. Methods in Molecular Biology, 2016, 1382: 21-39.
    [6] SAMULSKI R J, MUZYCZKA N. AAV-mediated gene therapy for research and therapeutic purposes[J]. Annual Review of Virology, 2014, 1(1): 427-451. doi: 10.1146/annurev-Virology-031413-085355
    [7] CARTER P J, SAMULSKI R J. Adeno-associated viral vectors as gene delivery vehicles[J]. International Journal of Molecular Medicine, 2000, 6(1): 17-27.
    [8] COLELLA P, RONZITTI G, MINGOZZI F. Emerging issues in AAV-mediatedin vivo gene Therapy[J]. Molecular Therapy Methods & Clinical Development, 2018, 8: 87-104.
    [9] LI C, BOWLES D E, VAN DYKE T, et al. Adeno-associated virus vectors: Potential applications for cancer gene therapy[J]. Cancer Gene Therapy, 2005, 12(12): 913-925. doi: 10.1038/sj.cgt.7700876
    [10] STILWELL J L, SAMULSKI R J. Adeno-associated virus vectors for therapeutic gene transfer[J]. BioTechniques, 2003, 34(1): 148-150. doi: 10.2144/03341dd01
    [11] CALCEDO R, MORIZONO H, WANG L, et al. Adeno-associated virus antibody profiles in newborns, children, and adolescents[J]. Clinical and Vaccine Immunology: CVI, 2011, 18(9): 1586-1588. doi: 10.1128/CVI.05107-11
    [12] CHEN Y H, CHANG M, DAVIDSON B L. Molecular signatures of disease brain endothelia provide new sites for CNS-directed enzyme therapy[J]. Nature Medicine, 2009, 15(10): 1215-1218. doi: 10.1038/nm.2025
    [13] PAULK N K, PEKRUN K, ZHU E, et al. Bioengineered AAV capsids with combined high human liver transduction in vivo and unique humoral seroreactivity[J]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2018, 26(1): 289-303. doi: 10.1016/j.ymthe.2017.09.021
    [14] MAHESHRI N, KOERBER J T, KASPAR B K, et al. Directed evolution of adeno-associated virus yields enhanced gene delivery vectors[J]. Nature Biotechnology, 2006, 24(2): 198-204. doi: 10.1038/nbt1182
    [15] KOERBER J T, JANG J H, SCHAFFER D V. DNA shuffling of adeno-associated virus yields functionally diverse viral progeny[J]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2008, 16(10): 1703-1709. doi: 10.1038/mt.2008.167
    [16] KOTTERMAN M A, SCHAFFER D V. Engineering adeno-associated viruses for clinical gene therapy[J]. Nature Reviews Genetics, 2014, 15(7): 445-451. doi: 10.1038/nrg3742
    [17] CHOUDHURY S R, FITZPATRICK Z, HARRIS A F, et al. In vivo selection yields AAV-B1 capsid for central nervous system and muscle gene therapy[J]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2016, 24(7): 1247-1257. doi: 10.1038/mt.2016.84
    [18] LI W, ZHANG L, JOHNSON J S, et al. Generation of novel AAV variants by directed evolution for improved CFTR delivery to human ciliated airway epithelium[J]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2009, 17(12): 2067-2077. doi: 10.1038/mt.2009.155
    [19] YANG L, JIANG J, DROUIN L M, et al. A myocardium tropic adeno-associated virus (AAV) evolved by DNA shuffling and in vivo selection[J]. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(10): 3946-3951. doi: 10.1073/pnas.0813207106
    [20] SANTIAGO-ORTIZ J, OJALA D S, WESTESSON O, et al. AAV ancestral reconstruction library enables selection of broadly infectious viral variants[J]. Gene Therapy, 2015, 22(12): 934-946. doi: 10.1038/gt.2015.74
    [21] ZINN E, PACOURET S, KHAYCHUK V, et al. In silico reconstruction of the viral evolutionary lineage yields a potent gene therapy vector[J]. Cell Reports, 2015, 12(6): 1056-1068. doi: 10.1016/j.celrep.2015.07.019
    [22] 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]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2014, 22(11): 1900-1909. doi: 10.1038/mt.2014.139
    [23] PERABO L, BUNING H, KOFLER D M, et al. In vitro selection of viral vectors with modified tropism: The adeno-associated virus display[J]. Molecular Therapy: The Journal of the American Society of Gene Therapy, 2003, 8(1): 151-157. doi: 10.1016/S1525-0016(03)00123-0
    [24] MULLER O J, KAUL F, WEITZMAN M D, et al. Random peptide libraries displayed on adeno-associated virus to select for targeted gene therapy vectors[J]. Nature Biotechnology, 2003, 21(9): 1040-1046. doi: 10.1038/nbt856
    [25] ZOLOTUKHIN S, BYRNE B J, MASON E, et al. Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield[J]. Gene Therapy, 1999, 6(6): 973-985. doi: 10.1038/sj.gt.3300938
    [26] SUN J, HUA B, CHEN X, et al. Gene delivery of activated factor VII using alternative adeno-associated virus serotype improves hemostasis in hemophiliac mice with FVIII inhibitors and adeno-associated virus neutralizing antibodies[J]. Human Gene Therapy, 2017, 28(8): 654-666. doi: 10.1089/hum.2017.016
    [27] FRONTERA W R, OCHALA J. Skeletal muscle: A brief review of structure and function[J]. Calcified Tissue International, 2015, 96(3): 183-195. doi: 10.1007/s00223-014-9915-y
    [28] KATTENHORN L M, TIPPER C H, STOICA L, et al. Adeno-associated virus gene therapy for liver disease[J]. Human Gene Therapy, 2016, 27(12): 947-961. doi: 10.1089/hum.2016.160
    [29] DEVERMAN B E, RAVINA B M, BANKIEWICZ K S, et al. Gene therapy for neurological disorders: Progress and prospects[J]. Nature Reviews Drug Discovery, 2018, 17(9): 641-659. doi: 10.1038/nrd.2018.110
    [30] MENDELL J R, AL-ZAIDY S, SHELL R, et al. Single-dose gene-replacement therapy for spinal muscular atrophy[J]. The New England Journal of Medicine, 2017, 377(18): 1713-1722. doi: 10.1056/NEJMoa1706198
    [31] FELSENTHAL N, ZELZER E. Mechanical regulation of musculoskeletal system development[J]. Development, 2017, 144(23): 4271-4283. doi: 10.1242/dev.151266
    [32] SANES J R, LICHTMAN J W. Induction, assembly, maturation and maintenance of a postsynaptic apparatus[J]. Nature Reviews Neuroscience, 2001, 2(11): 791-805. doi: 10.1038/35097557
    [33] MOUSE G S C, WATERSTON R H, LINDBLAD-TOH K, et al. Initial sequencing and comparative analysis of the mouse genome[J]. Nature, 2002, 420(6915): 520-562. doi: 10.1038/nature01262
    [34] FU H, MEADOWS A S, PINEDA R J, et al. Differential prevalence of antibodies against adeno-associated virus in healthy children and patients with mucopolysaccharidosis III: Perspective for AAV-mediated gene therapy[J]. Human Gene Therapy Clinical Development, 2017, 28(4): 187-196. doi: 10.1089/humc.2017.109
    [35] BOUTIN S, MONTEILHET V, VERON P, et al. Prevalence of serum IgG and neutralizing factors against adeno-associated virus (AAV) types 1, 2, 5, 6, 8, and 9 in the healthy population: Implications for gene therapy using AAV vectors[J]. Human Gene Therapy, 2010, 21(6): 704-712. doi: 10.1089/hum.2009.182
    [36] LIU Q, HUANG W, ZHANG H, et al. Neutralizing antibodies against AAV2, AAV5 and AAV8 in healthy and HIV-1-infected subjects in China: Implications for gene therapy using AAV vectors[J]. Gene Therapy, 2014, 21(8): 732-738. doi: 10.1038/gt.2014.47
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出版历程
  • 收稿日期:  2021-01-27
  • 网络出版日期:  2021-07-01
  • 刊出日期:  2022-02-23

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