Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

AAV2 serotype demonstrates the highest transduction efficiency in porcine lung-derived cells

  • Won Seok Ju (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Seokho Kim (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Areum Choi (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jae-Yeong Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Haesun Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jingu No (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Seunghoon Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Keonbong Oh (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jae Gyu Yoo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration)
  • Received : 2024.10.31
  • Accepted : 2024.12.11
  • Published : 2024.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Background: The ability of adeno-associated viruses (AAVs) to transduce various cell types with minimal immune responses renders them prominent vectors for gene editing (GE), with different AAV serotypes exhibiting distinct transduction efficiencies due to their specific cellular tropism. However, detailed molecular processes of AAV infection and penetration, as well as the optimal serotype for specific purposes, remain poorly understood. Porcine models are widely used in research benefitting both human and livestock due to anatomical and physiological similarities to humans. Methods: Transduction efficiencies of 18 AAV serotypes (AAV1-9, 6.2, rh10, DJ, DJ/8, PHP.eB, PHP.S, 2-retro, 2-QuadYF, and 2.7m8) were evaluated in immortalized porcine lung epithelial cells (pLCsImt) and pulmonary alveolar macrophages 3D4/31 (PAMs 3D4/31). Results: We found AAV2, DJ, and 2.7m8 to be the most effective in both cell types. The highest enhanced green fluorescent protein expression of 52.46 ± 2.4% in pLCsImt and 64.08 ± 2.4% in PAMs 3D4/31 was observed for AAV2, while negligible transduction was observed for AAV4, rh10, DJ, PHP.eB, PHP.S, and 2-retro. AAV-DJ showed superior transduction efficiency in PK-15, as compared to AAV2 and 2.7m8. Results emphasize the cell type-specific nature of AAV serotype transduction efficiencies. Notably, AAV2 was most effective in both lung and macrophage cells, whereas AAV-DJ was more effective in renal cells. Conclusions: Our findings suggest that AAV2 was identified as the most efficient serotype for transducing pLCsImt and PAMs 3D4/31, compare to the PK-15 cells. Understanding cell type-specific preferences of AAV serotypes offer crucial insight for tailoring AAV vectors to specific tissue and optimizing genome editing strategies, with potential implications for the advancement of personalized medicine and development of treatments for human and livestock.

Keywords

Acknowledgement

This study was supported by 2024 the RDA Fellowship Program of the National Institute of Animal Science, Rural Development Administration, Republic of Korea.

References

  1. Asmamaw Mengstie M. 2022. Viral vectors for the in vivo delivery of CRISPR components: advances and challenges. Front. Bioeng. Biotechnol. 12;10:895713. https://doi.org/10.3389/fbioe.2022.895713
  2. Balakrishnan B and Jayandharan GR. 2014. Basic biology of adeno-associated virus (AAV) vectors used in gene therapy. Curr. Gene Ther. 14:86-100. https://doi.org/10.2174/1566523214666140302193709
  3. Bennett A, Keravala A, Makal V, Kurian J, Belbellaa B, Aeran R, Tseng YS, Sousa D, Spear J, Gasmi M, Agbandje-McKenna M. 2020. Structure comparison of the chimeric AAV2.7m8 vector with parental AAV2. J. Struct. Biol. 209:107433. https://doi.org/10.1016/j.jsb.2019.107433
  4. Bijlani S, Pang KM, Sivanandam V, Singh A, Chatterjee S. 2022. The role of recombinant AAV in precise genome editing. Front. Genome Ed. 3:799722. https://doi.org/10.3389/fgeed.2021.799722
  5. Bähr A and Wolf E. 2012. Domestic animal models for biomedical research. Reprod. Domest. Anim. 47 Suppl 4:59-71. https://doi.org/10.1111/j.1439-0531.2012.02056.x
  6. Calcedo R, Morizono H, Wang L, McCarter R, He J, Jones D, Batshaw ML, Wilson JM. 2011. Adeno-associated virus antibody profiles in newborns, children, and adolescents. Clin. Vaccine. Immunol. 18:1586-1588. https://doi.org/10.1128/CVI.05107-11
  7. Calvert JG, Slade DE, Shields SL, Jolie R, Mannan RM, Ankenbauer RG, Welch SK. 2007. CD163 expression confers susceptibility to porcine reproductive and respiratory syndrome viruses. J. Virol. 81:7371-7379. https://doi.org/10.1128/JVI.00513-07
  8. Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sánchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V. 2017. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat. Neurosci. 20:1172-1179. https://doi.org/10.1038/nn.4593
  9. Chen CL, Jensen RL, Schnepp BC, Connell MJ, Shell R, Sferra TJ, Bartlett JS, Clark KR, Johnson PR. 2005. Molecular characterization of adeno-associated viruses infecting children. J. Virol. 79:14781-14792. https://doi.org/10.1128/JVI.79.23.14781-14792.2005
  10. Chen OG, Mather SE, Brommel CM, Hamilton BA, Ehler A, Villacreses R, Girgis RE, Abou Alaiwa M, Stoltz DA, Zabner J, Li X. 2021. Transduction of pig small airway epithelial cells and distal lung progenitor cells by AAV4. Cells 10:1014. https://doi.org/10.3390/cells10051014
  11. Chng K, Larsen SR, Zhou S, Wright JF, Martiniello-Wilks R, Rasko JE. 2007. Specific adeno-associated virus serotypes facilitate efficient gene transfer into human and non-human primate mesenchymal stromal cells. J. Gene Med. 9:22-32. https://doi.org/10.1002/jgm.990
  12. Cho IC, Park HB, Ahn JS, Han SH, Lee JB, Lim HT, Yoo CK, Jung EJ, Kim DH, Sun WS, Ramayo-Caldas Y, Kim SG, Kang YJ, Kim YK, Shin HS, Seong PN, Hwang IS, Park BY, Hwang S, Lee SS, Ryu YC, Lee JH, Ko MS, Lee K, Andersson G, Pérez Enciso M, Lee JW. 2019. A functional regulatory variant of MYH3 influences muscle fiber-type composition and intramuscular fat content in pigs. PLoS Genet 15:e1008279. https://doi.org/10.1371/journal.pgen.1008279
  13. Clauss S, Bleyer C, Schüttler D, Tomsits P, Renner S, Klymiuk N, Wakili R, Massberg S, Wolf E, Kääb S. 2019. Animal models of arrhythmia: classic electrophysiology to genetically modified large animals. Nat. Rev. Cardiol. 16:457-475. https://doi.org/10.1038/s41569-019-0179-0
  14. Cooney AL, Thornell IM, Singh BK, Shah VS, Stoltz DA, McCray PB Jr, Zabner J, Sinn PL. 2019. A novel AAV-mediated gene delivery system corrects CFTR function in pigs. Am. J. Respir. Cell Mol. Biol. 61:747-754. https://doi.org/10.1165/rcmb.2019-0006OC
  15. Dai Y, Kavita U, Lampen MH, Gielen S, Banks G, Levesque P, Kozhich A, Pillutla R, Zhang Y, Jawa V, Adam L. 2022. Prevalence of pre-existing neutralizing antibodies against adenoassociated virus serotypes 1, 2, 5, 6, 8, and 9 in sera of different pig strains. Hum. Gene Ther. 33:451-459. https://doi.org/10.1089/hum.2021.213
  16. Fan N and Lai L. 2013. Genetically modified pig models for human diseases. J. Genet. Genomics 40:67-73. https://doi.org/10.1016/j.jgg.2012.07.014
  17. Gaj T, Gersbach CA, Barbas CF 3rd. 2013. ZFN, TALEN, and CRISPR/Cas-based methods for genome engineering. Trends. Biotechnol. 31:397-405. https://doi.org/10.1016/j.tibtech.2013.04.004
  18. Gao G, Vandenberghe LH, Alvira MR, Lu Y, Calcedo R, Zhou X, Wilson JM. 2004. Clades of Adeno-associated viruses are widely disseminated in human tissues. J. Virol. 78:6381-6388. https://doi.org/10.1128/JVI.78.12.6381-6388.2004
  19. Gorovits B, Fiscella M, Havert M, Koren E, Long B, Milton M, Purushothama S. 2020. Recommendations for the development of cell-based anti-viral vector neutralizing antibody assays. AAPS J. 22:24. (Erratum published 2020, AAPS. J. 22:42). https://doi.org/10.1208/s12248-019-0403-1
  20. Grimm D, Lee JS, Wang L, Desai T, Akache B, Storm TA, Kay MA. 2008. In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adenoassociated viruses. J. Virol. 82:5887-5911. https://doi.org/10.1128/JVI.00254-08
  21. Halbert CL, Rutledge EA, Allen JM, Russell DW, Miller AD. 2000. Repeat transduction in the mouse lung by using adeno-associated virus vectors with different serotypes. J. Virol. 74:1524-1532. https://doi.org/10.1128/JVI.74.3.1524-1532.2000
  22. Haldrup SH, Fabian-Jessing BK, Jakobsen TS, Lindholm AB, Adsersen RL, Aagaard L, Bek T, Askou AL, Corydon TJ. 2024. Subretinal AAV delivery of RNAi-therapeutics targeting VEGFA reduces choroidal neovascularization in a large animal model. Mol. Ther. Methods Clin. Dev. 32:101242. https://doi.org/10.1016/j.omtm.2024.101242
  23. Hryhorowicz M, Zeyland J, Słomski R, Lipiński D. 2017. Genetically modified pigs as organ donors for xenotransplantation. Mol. Biotechnol. 59:435-444. https://doi.org/10.1007/s12033-017-0024-9
  24. Issa SS, Shaimardanova AA, Solovyeva VV, Rizvanov AA. 2023. Various AAV serotypes and their applications in gene therapy: an overview. Cells 12:785. https://doi.org/10.3390/cells12050785
  25. Kim S, No JG, Lee S, Choi A, Lee JY, Hyung N, Ju WS, Lee JY, Kwak TU, Lee P, Park MR, Byun SJ, Lee H, Oh KB, Yang H, Yoo JG. 2024. In vitro gene editing using primary cells derived from Cas9-expressing pigs. J. Anim. Sci. Technol. https://doi.org/10.5187/jast.2024.e77
  26. Kotterman MA, Yin L, Strazzeri JM, Flannery JG, Merigan WH, Schaffer DV. 2015. Antibody neutralization poses a barrier to intravitreal adeno-associated viral vector gene delivery to non-human primates. Gene Ther. 22:116-126. https://doi.org/10.1038/gt.2014.115
  27. Lisowski L, Tay SS, Alexander IE. 2015. Adeno-associated virus serotypes for gene therapeutics. Curr. Opin. Pharmacol. 24: 59-67. https://doi.org/10.1016/j.coph.2015.07.006
  28. Liu Z, Klose K, Neuber S, Jiang M, Gossen M, Stamm C. 2020. Comparative analysis of adeno-associated virus serotypes for gene transfer in organotypic heart slices. J. Transl. Med. 18:437. https://doi.org/10.1186/s12967-020-02605-4
  29. Logan GJ, Dane AP, Hallwirth CV, Smyth CM, Wilkie EE, Amaya AK, Zhu E, Khandekar N, Ginn SL, Liao SHY, Cunningham SC, Sasaki N, Cabanes-Creus M, Tam PPL, Russell DW, Lisowski L, Alexander IE. 2017. Identification of liver-specific enhancer-promoter activity in the 3' untranslated region of the wild-type AAV2 genome. Nat. Genet. 49:1267-1273. https://doi.org/10.1038/ng.3893
  30. Mendell JR, Al-Zaidy SA, Rodino-Klapac LR, Goodspeed K, Gray SJ, Kay CN, Boye SL, Boye SE, George LA, Salabarria S, Corti M, Byrne BJ, Tremblay JP. 2021. Current clinical applications of in vivo gene therapy with AAVs. Mol. Ther. 29:464-488. https://doi.org/10.1016/j.ymthe.2020.12.007
  31. Mingozzi F and High KA. 2011. Therapeutic in vivo gene transfer for genetic disease using AAV: progress and challenges. Nat. Rev. Genet. 12:341-355. (Erratum published 2011, Nat. Rev. Genet. 12:515). https://doi.org/10.1038/nrg2988
  32. Qi YF, Li QH, Shenoy V, Zingler M, Jun JY, Verma A, Katovich MJ, Raizada MK. 2013. Comparison of the transduction efficiency of tyrosine-mutant adeno-associated virus serotype vectors in kidney. Clin. Exp. Pharmacol. Physiol. 40:53-55. https://doi.org/10.1111/1440-1681.12037
  33. Senís E, Fatouros C, Große S, Wiedtke E, Niopek D, Mueller AK, Börner K, Grimm D. 2014. CRISPR/Cas9-mediated genome engineering: an adeno-associated viral (AAV) vector toolbox. Biotechnol. J. 9:1402-1412. https://doi.org/10.1002/biot.201400046
  34. Singh P and Ali SA. 2021. Impact of CRISPR-Cas9-based genome engineering in farm animals. Vet. Sci. 8:122. https://doi.org/10.3390/vetsci8070122
  35. Srivastava A. 2016. In vivo tissue-tropism of adeno-associated viral vectors. Curr. Opin. Virol. 21:75-80. https://doi.org/10.1016/j.coviro.2016.08.003
  36. Steines B, Dickey DD, Bergen J, Excoffon KJ, Weinstein JR, Li X, Yan Z, Abou Alaiwa MH, Shah VS, Bouzek DC, Powers LS, Gansemer ND, Ostedgaard LS, Engelhardt JF, Stoltz DA, Welsh MJ, Sinn PL, Schaffer DV, Zabner J. 2016. CFTR gene transfer with AAV improves early cystic fibrosis pig phenotypes. JCI Insight 1:e88728. https://doi.org/10.1172/jci.insight.88728
  37. Stender S, Murphy M, O'Brien T, Stengaard C, Ulrich-Vinther M, Søballe K, Barry F. 2007. Adeno-associated viral vector transduction of human mesenchymal stem cells. Eur. Cell Mater. 13:93-99; discussion 99. https://doi.org/10.22203/eCM.v013a10
  38. Su H, Yeghiazarians Y, Lee A, Huang Y, Arakawa-Hoyt J, Ye J, Orcino G, Grossman W, Kan YW. 2008. AAV serotype 1 mediates more efficient gene transfer to pig myocardium than AAV serotype 2 and plasmid. J. Gene Med. 10:33-41. https://doi.org/10.1002/jgm.1129
  39. Thornton PK. 2010. Livestock production: recent trends, future prospects. Philos. Trans. R Soc. Lond. B Biol. Sci. 365:2853-2867. https://doi.org/10.1098/rstb.2010.0134
  40. Van Gorp H, Van Breedam W, Delputte PL, Nauwynck HJ. 2008. Sialoadhesin and CD163 join forces during entry of the porcine reproductive and respiratory syndrome virus. J. Gen. Virol. 89(Pt 12):2943-2953. https://doi.org/10.1099/vir.0.2008/005009-0
  41. Wang K, Jin Q, Ruan D, Yang Y, Liu Q, Wu H, Zhou Z, Ouyang Z, Liu Z, Zhao Y, Zhao B, Zhang Q, Peng J, Lai C, Fan N, Liang Y, Lan T, Li N, Wang X, Wang X, Fan Y, Doevendans PA, Sluijter JPG, Liu P, Li X, Lai L. 2017. Cre-dependent Cas9-expressing pigs enable efficient in vivo genome editing. Genome Res. 27:2061-2071. https://doi.org/10.1101/gr.222521.117
  42. Wang Z, Maunze B, Wang Y, Tsoulfas P, Blackmore MG. 2018. Global connectivity and function of descending spinal input revealed by 3D microscopy and retrograde transduction. J. Neurosci. 38:10566-10581. https://doi.org/10.1523/JNEUROSCI.1196-18.2018
  43. Weber M, Rabinowitz J, Provost N, Conrath H, Folliot S, Briot D, Chérel Y, Chenuaud P, Samulski J, Moullier P, Rolling F. 2003. Recombinant adeno-associated virus serotype 4 mediates unique and exclusive long-term transduction of retinal pigmented epithelium in rat, dog, and nonhuman primate after subretinal delivery. Mol. Ther. 7:774-781. https://doi.org/10.1016/S1525-0016(03)00098-4
  44. Wells KD, Bardot R, Whitworth KM, Trible BR, Fang Y, Mileham A, Kerrigan MA, Samuel MS, Prather RS, Rowland RRR. 2017. Replacement of porcine CD163 scavenger receptor cysteine-rich domain 5 with a CD163-like homolog confers resistance of pigs to genotype 1 but not genotype 2 porcine reproductive and respiratory syndrome virus. J. Virol. 91:e01521-16. https://doi.org/10.1128/JVI.01521-16
  45. Whitworth KM, Green JA, Redel BK, Geisert RD, Lee K, Telugu BP, Wells KD, Prather RS. 2022. Improvements in pig agriculture through gene editing. CABI Agric. Biosci. 3:41. https://doi.org/10.1186/s43170-022-00111-9
  46. Whitworth KM, Rowland RR, Ewen CL, Trible BR, Kerrigan MA, Cino-Ozuna AG, Samuel MS, Lightner JE, McLaren DG, Mileham AJ, Wells KD, Prather RS. 2016. Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus. Nat. Biotechnol. 34:20-22. https://doi.org/10.1038/nbt.3434
  47. Wiley LA, Burnight ER, Kaalberg EE, Jiao C, Riker MJ, Halder JA, Luse MA, Han IC, Russell SR, Sohn EH, Stone EM, Tucker BA, Mullins RF. 2018. Assessment of adeno-associated virus serotype tropism in human retinal explants. Hum. Gene Ther. 29:424-436. https://doi.org/10.1089/hum.2017.179
  48. Willimann M, Tiyaboonchai A, Adachi K, Li B, Waldburger L, Nakai H, Grompe M, Thöny B. 2024. AAV capsid screening for translational pig research using a mouse xenograft liver model. bioRxiv. https://doi.org/10.1101/2024.05.29.596409
  49. Yoon DS, Lee KM, Cho S, Ko EA, Kim J, Jung S, Shim JH, Gao G, Park KH, Lee JW. 2021. Cellular and tissue selectivity of AAV serotypes for gene delivery to chondrocytes and cartilage. Int. J. Med. Sci. 18:3353-3360. https://doi.org/10.7150/ijms.56760
  50. Zengel J, Wang YX, Seo JW, Ning K, Hamilton JN, Wu B, Raie M, Holbrook C, Su S, Clements DR, Pillay S, Puschnik AS, Winslow MM, Idoyaga J, Nagamine CM, Sun Y, Mahajan VB, Ferrara KW, Blau HM, Carette JE. 2023. Hardwiring tissuespecific AAV transduction in mice through engineered receptor expression. Nat. Methods. 20:1070-1081. https://doi.org/10.1038/s41592-023-01896-x
  51. Zhang J, Wang J, Zhang X, Zhao C, Zhou S, Du C, Tan Y, Zhang Y, Shi K. 2022. Transcriptome profiling identifies immune response genes against porcine reproductive and respiratory syndrome virus and Haemophilus parasuis co-infection in the lungs of piglets. J. Vet. Sci. 23:e2. https://doi.org/10.4142/jvs.2022.23.e2
  52. Zincarelli C, Soltys S, Rengo G, Rabinowitz JE. 2008. Analysis of AAV serotypes 1-9 mediated gene expression and tropism in mice after systemic injection. Mol. Ther. 16:1073-1080. https://doi.org/10.1038/mt.2008.76