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AAV3-miRNA vectors for growth suppression of human hepatocellular carcinoma cells in vitro and human liver tumors in a murine xenograft model in vivo

Gene Therapy volume 28, pages 422–434 (2021)Cite this article

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Abstract

We have previously reported that recombinant adeno-associated virus serotype 3 (AAV3) vectors transduce human liver tumors more efficiently in a mouse xenograft model following systemic administration. Others have utilized AAV8 vectors expressing miR-26a and miR-122 to achieve near total inhibition of growth of mouse liver tumors. Since AAV3 vectors transduce human hepatic cells more efficiently than AAV8 vectors, in the present studies, we wished to evaluate the efficacy of AAV3-miR-26a/122 vectors in suppressing the growth of human hepatocellular carcinoma (HCC) cells in vitro, and human liver tumors in a mouse model in vivo. To this end, a human HCC cell line, Huh7, was transduced with various multiplicities of infection (MOIs) of AAV3-miR-26a or scAAV3-miR-122 vectors, or both, which also co-expressed a Gaussia luciferase (GLuc) reporter gene. Only a modest level of dose-dependent growth inhibition of Huh7 cells (~12–13%) was observed at the highest MOI (1 × 105 vgs/cell) with each vector. When Huh7 cells were co-transduced with both vectors, the extent of growth inhibition was additive (~26%). However, AAV3-miR-26a and scAAV3-miR-122 vectors led to ~70% inhibition of growth of Huh-derived human liver tumors in a mouse xenograft model in vivo. Thus, the combined use of miR-26a and scAAV3-miR-122 delivered by AAV3 vectors offers a potentially useful approach to target human liver tumors.

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Fig. 1: Construction and characterization of scAAV3 vectors.
Fig. 2: scAAV3-miRNA vectors-mediated growth inhibition of Huh7 human hepatocellular carcinoma cells.
Fig. 3: scAAV3-miRNA vectors-mediated growth inhibition of Huh7 cells stably expressing the firefly luciferase (FLuc) reporter gene.
Fig. 4: scAAV3-miRNA vectors-mediated growth inhibition of Huh7-FLuc cells correlates with diminution of transgene expression and loss of viral vector genomes as a function of time.
Fig. 5: Loss of vector genome copy numbers correlates with expression levels of miRNAs in Huh7-FLuc cells.
Fig. 6: Growth inhibition of Huh7-FLuc cells with scAAV3-miRNA vectors is additive, and correlates with diminution of transgene expression and loss of viral vector genomes.
Fig. 7: Lack of growth inhibition of HepaRG cells by scAAV3-miRNA vectors.
Fig. 8: Suppression of growth of tumors induced by Huh7-FLuc cells transduced with scAAV3-miRNA vectors in vivo.
Fig. 9: Suppression of growth of tumors induced by Huh7-FLuc cells transduced with scAAV3-miRNA vectors in vivo as a function of time.
Fig. 10: Lack of effect of tumor growth on the average weight of NSG mice.

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References

  1. Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2018;68:394–424.

    Article  PubMed  Google Scholar 

  2. Crissien AM, Frenette C. Current management of hepatocellular carcinoma. Gastroenterol Hepatol. 2014;10:153–61.

    Google Scholar 

  3. Wang X, Wang N, Cheung F, Lao L, Li C, Feng Y. Chinese medicines for prevention and treatment of human hepatocellular carcinoma: current progress on pharmacological actions and mechanisms. J Integr Med. 2015;13:142–64.

    Article  PubMed  Google Scholar 

  4. Forner A, Llovet JM, Bruix J. Hepatocellular carcinoma. Lancet. 2012;379:1245–55.

    Article  Google Scholar 

  5. Greene CM, Varley RB, Lawless MW. MicroRNAs and liver cancer associated with iron overload: therapeutic targets unravelled. World J Gastroenterol. 2013;19:5212–26.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Erridge S, Pucher PH, Markar SR, Malietzis G, Athanasiou T, Darzi A, et al. Meta-analysis of determinants of survival following treatment of recurrent hepatocellular carcinoma. Br J Surg. 2017;104:1433–42.

    Article  CAS  PubMed  Google Scholar 

  7. Ling CQ, Fan J, Lin HS, Shen F, Xu ZY, Lin LZ, et al. Clinical practice guidelines for the treatment of primary liver cancer with integrative traditional Chinese and Western medicine. J Integr Med. 2018;16:236–48.

    Article  PubMed  Google Scholar 

  8. Lee RC, Feinbaum RL, Ambrost V. The C. Elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-4. Cell. 1993;75:843–54.

    Article  CAS  PubMed  Google Scholar 

  9. Bartel DP. MicroRNAs: Target recognition and regulatory functions. Cell. 2009;136:215–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Port JD, Sucharov C. Role of microRNAs in cardiovascular disease: therapeutic challenges and potentials. J Cardiovasc Pharmacol. 2010;56:444–53.

    Article  CAS  PubMed  Google Scholar 

  11. Pager CT, Wehner KA, Fuchs G, Sarnow P. MicroRNA-mediated gene silencing. Prog Mol Biol Transl Sci. 2009;90:187–210.

    Article  CAS  PubMed  Google Scholar 

  12. Hammond SM. MicroRNA therapeutics: a new niche for antisense nucleic acids. Trends Mol Med. 2006;12:99–101.

    Article  CAS  PubMed  Google Scholar 

  13. Chen J, Zhang K, Xu YJ, Gao YP, Li C, Wang R, et al. The role of microRNA-26a in human cancer progression and clinical application. Tumor Biol. 2016;37:7095–108.

    Article  CAS  Google Scholar 

  14. Sartorius K, Sartorius B, Cheryl W, Chuturgoon A, Makarova J. The biological and diagnostic role of mirna’s in hepatocellular carcinoma. Front Biosci. 2018;23:1701–20.

    Article  CAS  Google Scholar 

  15. Yang X, Liang L, Zhang XF, Jia HL, Qin Y, Zhu XC, et al. MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin-6-Stat3 pathway. Hepatology. 2013;58:158–70.

    Article  CAS  PubMed  Google Scholar 

  16. Chai Z, Kong J, Zhu X, Zhang Y, Lu L, Zhou J, et al. MicroRNA-26a inhibits angiogenesis by down-regulating VEGFA through the PIK3C2α/Akt/HIF-1α pathway in hepatocellular carcinoma. PLoS ONE. 2013;8:e77957.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Yang X, Zhang XF, Lu X, Jia HL, Liang L, Dong QZ, et al. MicroRNA-26a suppresses angiogenesis in human hepatocellular carcinoma by targeting hepatocyte growth factor-cMet pathway. Hepatology. 2014;59:1874–85.

    Article  CAS  PubMed  Google Scholar 

  18. Ji JF, Shi J, Budhu A, Yu ZP, Forgues M, Roessler S, et al. MicroRNA expression, survival, and response to interferon in liver cancer. N Engl J Med. 2009;361:1437–47.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lagos-Quintana M, Rauhut R, Yalcin A, Meyer J, Lendeckel W, Tuschl T. Identification of tissue-specific microRNAs from mouse. Curr Biol. 2002;12:735–9.

    Article  CAS  PubMed  Google Scholar 

  20. Xu J, Zhu XM, Wu LJ, Yang R, Yang ZR, Wang QF, et al. microRNA-122 suppresses cell proliferation and induces cell apoptosis in hepatocellular carcinoma by directly targeting Wnt/β-catenin pathway. Liver Int. 2012;32:752–60.

    Article  CAS  PubMed  Google Scholar 

  21. Gramantieri L, Ferracin M, Fornari F, Veronese A, Sabbioni S, Liu CG, et al. Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma. Cancer Res. 2007;67:6092–9.

    Article  CAS  PubMed  Google Scholar 

  22. Wang B, Hsu SH, Wang XM, Kutay H, Bid HK, Yu JH, et al. Reciprocal regulation of microRNA-122 and c-Myc in hepatocellular cancer: role of E2F1 and TFDP2. Hepatology. 2014;59:555–66.

    Article  CAS  PubMed  Google Scholar 

  23. Fan CG, Wang CM, Tian C, Wang Y, Li L, Sun WS, et al. miR-122 inhibits viral replication and cell proliferation in hepatitis B virus-related hepatocellular carcinoma and targets NDRG3. Oncol Rep. 2011;26:1281–6.

    CAS  PubMed  Google Scholar 

  24. Sun J, Lu HQ, Wang X, Jin HC. MicroRNAs in hepatocellular carcinoma: regulation, function, and clinical implications. Sci World J. 2013;2013:1–14.

    Google Scholar 

  25. Tsai WC, Hsu PW, Lai TC, Chau GY, Lin CW, Chen CM, et al. MicroRNA-122, a tumor suppressor microRNA that regulates intrahepatic metastasis of hepatocellular carcinoma. Hepatology. 2009;49:1571–82.

    Article  CAS  PubMed  Google Scholar 

  26. Li MX, Jayandharan GR, Li BZ, Ling C, Ma WQ, Srivastava A, et al. High-Efficiency transduction of fibroblasts and mesenchymal stem cells by Tyrosine-Mutant AAV2 vectors for their potential use in cellular therapy. Hum Gene Ther. 2010;21:1527–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Kota J, Chivukula RR, O’Donnell KA, Wentzel EA, Montgomery CL, Hwang HW, et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model. Cell. 2009;137:1005–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Hsu SH, Wang B, Kota J, Yu JH, Costinean S, Kutay H, et al. Essential metabolic, anti-inflammatory, and anti-tumorigenic functions of miR-122 in liver. J Clin Investig. 2012;122:2871–83.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Bainbridge JW, Smith AJ, Barker SS, Robbie S, Henderson R, Balaggan K, et al. Effect of gene therapy on visual function in Leber’s congenital amaurosis. N Engl J Med. 2008;358:2231–9.

    Article  CAS  PubMed  Google Scholar 

  30. Maguire AM, Simonelli F, Pierce EA, Pugh EN Jr, Mingozzi F, Bennicelli J, et al. Safety and efficacy of gene transfer for Leber’s congenital amaurosis. N Engl J Med. 2008;358:2240–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Hauswirth WW, Aleman TS, Kaushal S, Cideciyan AV, Schwartz SB, Wang L, et al. Treatment of Leber congenital amaurosis due to RPE65 mutations by ocular subretinal injection of adeno–associated virus gene vector: short-term results of a phase I trial. Hum Gene Ther. 2008;19:979–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Cideciyan AV, Aleman TS, Boye SL, Schwartz SB, Kaushal S, Roman AJ, et al. Human gene therapy for RPE65 isomerase deficiency activates the retinoid cycle of vision but with slow rod kinetics. Proc Natl Acad Sci USA. 2008;105:15112–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Gaudet D, Méthot J, Déry S, Brisson D, Essiembre C, Tremblay G, et al. Efficacy and long-term safety of alipogene tiparvovec (AAV1-LPLS447X) gene therapy for lipoprotein lipase deficiency: an open-label trial. Gene Ther. 2013;20:361–9.

    Article  CAS  PubMed  Google Scholar 

  34. Nathwani AC, Tuddenham EG, Rangarajan S, Rosales C, McIntosh J, Linch DC, et al. Adenovirus-associated virus vector-mediated gene transfer in Hemophilia B. N Engl J Med. 2011;365:2357–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Nathwani AC, Reiss UM, Tuddenham EG, Rosales C, Chowdary P, McIntosh J, et al. Long-term safety and efficacy of Factor IX gene therapy in Hemophilia B. N Engl J Med. 2014;371:1994–2004.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  36. Miesbach W, Meijer K, Coppens M, Kampmann P, Klamroth R, Schutgens R, et al. Gene therapy with adeno-associated virus vector 5-human Factor IX in adults with Hemophilia B. Blood. 2018;131:1022–31.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. George LA, Sullivan SK, Giermasz A, Rasko JEJ, Samelson-Jones BJ, Ducore J, et al. Hemophilia B gene therapy with a high-specific-activity Factor IX variant. N Engl J Med. 2017;377:2215–27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Rangarajan S, Walsh L, Lester W, Perry D, Madan B, Laffan M, et al. AAV5-Factor VIII gene transfer in severe Hemophilia A. N Engl J Med. 2017;377:2519–30.

    Article  CAS  PubMed  Google Scholar 

  39. Pasi KJ, Rangarajan S, Mitchell N, Lester W, Symington E, Madan B, et al. Multiyear follow-up of AAV5-hFVIII-SQ gene therapy for Hemophilia A. N Engl J Med. 2020;382:29–40.

    Article  CAS  PubMed  Google Scholar 

  40. Hwu WL, Muramatsu S, Tseng SH, Tzen KY, Lee NC, Chien YH, et al. Gene therapy for aromatic L-amino acid decarboxylase deficiency. Sci Transl Med. 2012;4:134ra61.

    Article  PubMed  CAS  Google Scholar 

  41. MacLaren RE, Groppe M, Barnard AR, Cottriall CL, Tolmachova T, Seymour L, et al. Retinal gene therapy in patients with choroideremia: initial findings from a phase 1/2 clinical trial. Lancet. 2014;383:1129–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Guy J, Feuer WJ, Davis JL, Porciatti V, Gonzalez PJ, Koilkonda RD, et al. Gene therapy for Leber hereditary optic neuropathy: low- and medium-dose visual results. Ophthalmology. 2017;124:1621–34.

    Article  PubMed  Google Scholar 

  43. Mendell JR, Al-Zaidy S, Shell R, Arnold WD, Rodino-Klapac LR, Prior TW, et al. Single-dose gene-replacement therapy for spinal muscular atrophy. N Engl J Med. 2017;377:1713–22.

    Article  CAS  PubMed  Google Scholar 

  44. Ling C, Lu Y, Cheng B, McGoogan KE, Gee SWY, Ma WQ, et al. High-Efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors. J Vis Exp. 2011;49:2538.

    Google Scholar 

  45. Cheng BB, Ling C, Dai Y, Lu Y, Glushakova LG, Gee SWY, et al. Development of optimized AAV3 serotype vectors: mechanism of high-efficiency transduction of human liver cancer cells. Gene Ther. 2012;19:375–84.

    Article  CAS  PubMed  Google Scholar 

  46. Ling C, Wang Y, Zhang YH, Ejjigani A, Yin ZF, Lu Y, et al. Selective in vivo targeting of human liver tumors by optimized AAV3 vectors in a murine xenograft model. Hum Gene Ther. 2014;25:1023–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Liu Y, Joo KI, Wang P. Endocytic processing of adeno-associated virus type 8 vectors for transduction of target cells. Gene Ther. 2013;20:308–17.

    Article  CAS  PubMed  Google Scholar 

  48. Kube DM, Srivastava A. Quantitative DNA slot blot analysis: inhibition of DNA binding to membranes by magnesium ions. Nucleic Acids Res. 1997;25:3375–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Ling C, Wang Y, Lu Y, Wang LN, Jayandharan GR, Aslanidi GV, et al. Enhanced transgene expression from recombinant single-stranded D-sequence-substituted adeno-associated virus vectors in human cell lines in vitro and in murine hepatocytes in vivo. J Virol. 2015;89:952–61.

    Article  PubMed  CAS  Google Scholar 

  50. Aslanidi GV, Rivers AE, Ortiz L, Song L, Ling C, Govindasamy L, et al. Optimization of the capsid of recombinant adeno-associated virus 2 (AAV2) vectors: the final threshold? PLoS ONE. 2013;8:e59142.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Nakabayashi H, Taketa K, Miyano K, Yamane T, Sato J. Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Res. 1982;42:3858–63.

    CAS  PubMed  Google Scholar 

  52. Nowrouzi A, Penaud-Budloo M, Kaeppel C, Appelt U, Le Guiner C, Moullier P, et al. Integration frequency and intermolecular recombination of rAAV vectors in non-human primate skeletal muscle and liver. Mol Ther. 2012;20:1177–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Guillouzo A, Corlu A, Aninat C, Glaise D, Morel F, Guguen-Guillouzo C. The human hepatoma HepaRG cells: a highly differentiated model for studies of liver metabolism and toxicity of xenobiotics. Chem Biol Interact. 2007;168:66–73.

    Article  CAS  PubMed  Google Scholar 

  54. Glushakova LG, Lisankie MJ, Eruslanov EB, Ojano-Dirain C, Zolotukhin I, Liu C, et al. AAV3-mediated transfer and expression of the pyruvate dehydrogenase E1 alpha subunit gene causes metabolic remodeling and apoptosis of human liver cancer cells. Mol Genet Metab. 2009;98:289–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Ling C, Li BZ, Ma WQ, Srivastava A. Development of optimized AAV serotype vectors for high-efficiency transduction at further reduced doses. Hum Gene Ther Method. 2016;27:143–9.

    Article  CAS  Google Scholar 

  56. Zhang YH, Wang Y, Yusufali AH, Ashby F, Zhang D, Yin ZF, et al. Cytotoxic genes from traditional Chinese medicine inhibit tumor growth both in vitro and in vivo. J Integr Med. 2014;12:483–94.

    Article  PubMed  Google Scholar 

  57. Qadir MI, Rizvi SZ. MiRNA in hepatocellular carcinoma: Pathogenesis and therapeutic approaches. Crit Rev Eukaryot Gene Expr. 2017;27:355–61.

    Article  PubMed  Google Scholar 

  58. Bandiera S, Pfeffer S, Baumert TF, Zeisel MB. miR-122—a key factor and therapeutic target in liver disease. J Hepatol. 2015;62:448–57.

    Article  CAS  PubMed  Google Scholar 

  59. Janssen HLA, Reesink HW, Lawitz EJ, Zeuzem S, Rodriguez-Torres M, Patel K, et al. Treatment of HCV infection by targeting microRNA. N Engl J Med. 2013;368:1685–94.

    Article  CAS  PubMed  Google Scholar 

  60. Chang L, Li K, Guo T. miR-26a-5p suppresses tumor metastasis by regulating EMT and is associated with prognosis in HCC. Clin Transl Oncol. 2017;19:695–703.

    Article  CAS  PubMed  Google Scholar 

  61. Fukuhara T, Kambara H, Shiokawa M, Ono C, Katoh H, Morita E, et al. Expression of microRNA miR-122 facilitates an efficient replication in nonhepatic cells upon infection with hepatitis C virus. J Virol. 2012;86:7918–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  62. Mizuguchi Y, Takizawa T, Yoshida H, Uchida E. Dysregulated miRNA in progression of hepatocellular carcinoma: a systematic review. Hepatol Res. 2016;46:391–406.

    Article  CAS  PubMed  Google Scholar 

  63. Yin J, Tang HF, Xiang Q, Yu J, Yang XY, Hu N, et al. MiR-122 increases sensitivity of drug-resistant BEL-7402/5-FU cells to 5-fluorouracil via down-regulation of bcl-2 family proteins. Pharmazie. 2011;66:975–81.

    CAS  PubMed  Google Scholar 

  64. Gao G, Alvira MR, Somanathan S, Lu Y, Vandenberghe LH, Rux JJ, et al. Adeno-associated viruses undergo substantial evolution in primates during natural infections. Proc Natl Acad Sci USA. 2003;100:6081–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Sarkar R, Tetreault R, Gao G, Wang L, Bell P, Chandler R, et al. Total correction of Hemophilia A mice with canine FVIII using an AAV 8 serotype. Blood. 2004;103:1253–60.

    Article  CAS  PubMed  Google Scholar 

  66. Li S, Ling C, Zhong L, Li M, Su Q, He R, et al. Efficient and targeted transduction of nonhuman primate liver with systemically delivered optimized AAV3B vectors. Mol Ther. 2015;23:1867–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Wang L, Bell P, Somanathan S, Wang Q, He Z, Yu H, et al. Comparative study of liver gene transfer with AAV vectors based on natural and engineered AAV capsids. Mol Ther. 2015;23:1877–87.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Vercauteren K, Hoffman BE, Zolotukhin I, Keeler GD, Xiao JW, Basner-Tschakarjan E, et al. Superior in vivo transduction of human hepatocytes using engineered AAV3 capsid. Mol Ther. 2016;24:1042–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  69. El-Serag HB. Current status of Sorafenib use for treatment of hepatocellular carcinoma. Gastroenterol Hepatol. 2017;13:623–5.

    Google Scholar 

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Acknowledgements

This research was supported in part by Public Health Service grants R01 HL-097088, R41 AI-122735, and R21 EB-015684 from the National Institutes of Health; a grant from the Children’s Miracle Network; and support from the Kitzman Foundation (to AS). LY was supported by the International Postdoctoral Exchange Fellowship Program 2018 by the Office of China Postdoctoral Council.

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  1. Yuanhui Zhang

    Present address: Department of Oncology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, China

Authors and Affiliations

  1. State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai, China

    Ling Yin, Yuanhui Zhang & Chen Ling

  2. Division of Cellular and Molecular Therapy, Department of Pediatrics, University of Florida College of Medicine, Gainesville, FL, USA

    Ling Yin, Geoffrey D. Keeler, Yuanhui Zhang, Brad E. Hoffman, Chen Ling, Keyun Qing & Arun Srivastava

  3. Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, FL, USA

    Ling Yin, Geoffrey D. Keeler, Yuanhui Zhang, Brad E. Hoffman, Chen Ling, Keyun Qing & Arun Srivastava

  4. Department of Molecular Genetics & Microbiology, University of Florida College of Medicine, Gainesville, FL, USA

    Chen Ling & Arun Srivastava

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Contributions

LY, GDK, YZ, KQ performed the experiments. LY, GDK, YZ, BEH, CL, KQ, and AS analyzed the data. CL and AS conceived of the idea. LY and AS wrote the paper, and all authors read and approved the final version.

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Correspondence to Chen Ling or Arun Srivastava.

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AS is a cofounder of, and holds equity in, Lacerta Therapeutics, aaVective, KASHX Bio, and Nirvana Therapeutics, and is an inventor on several issued patents on rAAV vectors that have been licensed to various gene therapy companies. All other authors declare no conflict of interests.

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Yin, L., Keeler, G.D., Zhang, Y. et al. AAV3-miRNA vectors for growth suppression of human hepatocellular carcinoma cells in vitro and human liver tumors in a murine xenograft model in vivo. Gene Ther 28, 422–434 (2021). https://doi.org/10.1038/s41434-020-0140-1

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