Skip to main content

Advertisement

SpringerLink
Log in

Utilization of a novel Sendai virus vector in ex vivo gene therapy for hemophilia A

  • Original Article
  • Published:
International Journal of Hematology Aims and scope Submit manuscript

Abstract

Sendai virus (SeV) vectors are being recognized as a superior tool for gene transfer. Here, we report the transfection efficacy of a novel, high-performance, replication-defective, and persistent Sendai virus (SeVdp) vector in cultured cells and in mice using a near-infrared fluorescent protein (iRFP)-mediated in vivo imaging system. The novel SeVdp vector established persistent infection, and strong expression of inserted genes was sustained indefinitely in vitro. Analysis of iRFP-expressing cells transplanted subcutaneously into NOG, nude, and ICR mice suggests that innate immunity was involved in the exclusion of the transplanted cells. We also evaluated the feasibility of this novel SeVdp vector for hemophilia A gene therapy. This system enabled insertion of full-length FVIII genes, and transduced cells secreted FVIII into the culture medium. Transient FVIII activity was detected in the plasma of mice after intraperitoneal transplantation of these FVIII-secreting cells. Further improvement in methods to evade immunity, such as simultaneous expression of immunomodulatory genes, would make this novel vector a very useful tool in regenerative medicine.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Nakanishi M, Otsu M. Development of Sendai virus vectors and their potential applications in gene therapy and regenerative medicine. Curr Gene Ther. 2012;12:410–6.

    Article  CAS  Google Scholar 

  2. Hurwitz JL. Development of recombinant Sendai virus vaccines for prevention of human parainfluenza and respiratory syncytial virus infections. Pediatr Infect Dis J. 2008;27:S126–8.

    Article  Google Scholar 

  3. Masaki I, Yonemitsu Y, Yamashita A, Sata S, Tanii M, Komori K, et al. Angiogenic gene therapy for experimental critical limb ischemia: acceleration of limb loss by overexpression of vascular endothelial growth factor 165 but not of fibroblast growth factor-2. Circ Res. 2002;90:966–73.

    Article  CAS  Google Scholar 

  4. Hasegawa Y, Kinoh H, Iwadate Y, Onimaru M, Ueda Y, Harada Y, et al. Urokinase-targeted fusion by oncolytic Sendai virus eradicates orthotopic glioblastomas by pronounced synergy with interferon-beta gene. Mol Ther. 2010;18:1778–86.

    Article  CAS  Google Scholar 

  5. Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y, et al. Development of defective and persistent Sendai virus vector: a unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem. 2011;286:4760–71.

    Article  CAS  Google Scholar 

  6. Yoshida T, Nagai Y, Maeno K, Iinuma M, Hamaguchi M, Matsumoto T, et al. Studies on the role of M protein in virus assembly using a ts mutant of HVJ (Sendai virus). Virology. 1979;92:139–54.

    Article  CAS  Google Scholar 

  7. Eguchi A, Kondoh T, Kosaka H, Suzuki T, Momota H, Masago A, et al. Identification and characterization of cell lines with a defect in a post-adsorption stage of Sendai virus-mediated membrane fusion. J Biol Chem. 2000;275:17549–55.

    Article  CAS  Google Scholar 

  8. Manco-Johnson MJ, Abshire TC, Shapiro AD, Riske B, Hacker MR, Kilcoyne R, et al. Prophylaxis versus episodic treatment to prevent joint disease in boys with severe hemophilia. N Engl J Med. 2007;357:535–44.

    Article  CAS  Google Scholar 

  9. Oldenburg J, Mahlangu JN, Kim B, Schmitt C, Callaghan MU, Young G, et al. Emicizumab prophylaxis in hemophilia A with inhibitors. N Engl J Med. 2017;377:809–18.

    Article  CAS  Google Scholar 

  10. Matsui H, Shibata M, Brown B, Labelle A, Hegadorn C, Andrews C, et al. Ex vivo gene therapy for hemophilia A that enhances safe delivery and sustained in vivo factor VIII expression from lentivirally engineered endothelial progenitors. Stem Cells. 2007;25:2660–9.

    Article  CAS  Google Scholar 

  11. Ohmori T, Mizukami H, Ozawa K, Sakata Y, Nishimura S. New approaches to gene and cell therapy for hemophilia. J Thromb Haemost. 2015;13(Suppl 1):S133–42.

    Article  Google Scholar 

  12. 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  Google Scholar 

  13. Louis Jeune V, Joergensen JA, Hajjar RJ, Weber T. Pre-existing anti-adeno-associated virus antibodies as a challenge in AAV gene therapy. Hum Gene Ther Methods. 2013;24:59–67.

    Article  CAS  Google Scholar 

  14. Wu Z, Yang H, Colosi P. Effect of genome size on AAV vector packaging. Mol Ther. 2010;18:80–6.

    Article  CAS  Google Scholar 

  15. Brown HC, Wright JF, Zhou S, Lytle AM, Shields JE, Spencer HT, et al. Bioengineered coagulation factor VIII enables long-term correction of murine hemophilia A following liver-directed adeno-associated viral vector delivery. Mol Ther Methods Clin Dev. 2014;1:14036.

    Article  Google Scholar 

  16. Hirsch ML, Wolf SJ, Samulski RJ. Delivering transgenic DNA exceeding the carrying capacity of AAV vectors. Methods Mol Biol. 2016;1382:21–39.

    Article  CAS  Google Scholar 

  17. Kaufman RJ, Pipe SW, Tagliavacca L, Swaroop M, Moussalli M. Biosynthesis, assembly and secretion of coagulation factor VIII. Blood Coagul Fibrinolysis. 1997;8(Suppl 2):S3-14.

    CAS  PubMed  Google Scholar 

  18. Fomin ME, Togarrati PP, Muench MO. Progress and challenges in the development of a cell-based therapy for hemophilia A. J Thromb Haemost. 2014;12:1954–65.

    Article  CAS  Google Scholar 

  19. Matsui H, Fujimoto N, Sasakawa N, Ohinata Y, Shima M, Yamanaka S, et al. Delivery of full-length factor VIII using a piggyBac transposon vector to correct a mouse model of hemophilia A. PLoS ONE. 2014;9:e104957.

    Article  Google Scholar 

  20. Shi Q. Platelet-targeted gene therapy for hemophilia. Mol Ther Methods Clin Dev. 2018;9:100–8.

    Article  CAS  Google Scholar 

  21. Ito M, Bujo H, Takahashi K, Arai T, Tanaka I, Saito Y. Implantation of primary cultured adipocytes that secrete insulin modifies blood glucose levels in diabetic mice. Diabetologia. 2005;48:1614–20.

    Article  CAS  Google Scholar 

  22. Aoyagi Y, Kuroda M, Asada S, Bujo H, Tanaka S, Konno S, et al. Fibrin glue increases the cell survival and the transduced gene product secretion of the ceiling culture-derived adipocytes transplanted in mice. Exp Mol Med. 2011;43:161–7.

    Article  CAS  Google Scholar 

  23. Spalding KL, Arner E, Westermark PO, Bernard S, Buchholz BA, Bergmann O, et al. Dynamics of fat cell turnover in humans. Nature. 2008;453:783–7.

    Article  CAS  Google Scholar 

  24. Shimizu T, Ishikawa T, Sugihara E, Kuninaka S, Miyamoto T, Mabuchi Y, et al. c-MYC overexpression with loss of Ink4a/Arf transforms bone marrow stromal cells into osteosarcoma accompanied by loss of adipogenesis. Oncogene. 2010;29:5687–99.

    Article  CAS  Google Scholar 

  25. Miao HZ, Sirachainan N, Palmer L, Kucab P, Cunningham MA, Kaufman RJ, et al. Bioengineering of coagulation factor VIII for improved secretion. Blood. 2004;103:3412–9.

    Article  CAS  Google Scholar 

  26. Filonov GS, Piatkevich KD, Ting LM, Zhang J, Kim K, Verkhusha VV. Bright and stable near-infrared fluorescent protein for in vivo imaging. Nat Biotechnol. 2011;29:757–61.

    Article  CAS  Google Scholar 

  27. Tran MT, Tanaka J, Hamada M, Sugiyama Y, Sakaguchi S, Nakamura M, et al. In vivo image analysis using iRFP transgenic mice. Exp Anim. 2014;63:311–9.

    Article  CAS  Google Scholar 

  28. Jobsis FF. Noninvasive, infrared monitoring of cerebral and myocardial oxygen sufficiency and circulatory parameters. Science. 1977;198:1264–7.

    Article  CAS  Google Scholar 

  29. Lai CW, Chen HL, Yen CC, Wang JL, Yang SH, Chen CM. Using dual fluorescence reporting genes to establish an in vivo imaging model of orthotopic lung adenocarcinoma in mice. Mol Imaging Biol. 2016;18:849–59.

    Article  CAS  Google Scholar 

  30. Neumeyer J, Lin RZ, Wang K, Hong X, Hua T, Croteau SE, et al. Bioengineering hemophilia A-specific microvascular grafts for delivery of full-length factor VIII into the bloodstream. Blood Adv. 2019;3:4166–76.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by the following grants: Health Labour Sciences Research Grant (GA26096), Research Project for Practical Applications of Regenerative Medicine (JP16bk0104032) by Japan Agency for Medical Research and Development (AMED), Grant-in-Aid for Scientific Research (KAKENHI) (26670264, 26221004, 15H04281, 16H01630, 16K14589), Industry-University collaboration project of University of Tsukuba (15-1), and AMED Grant Number JP17lm0203010. The authors would like to thank Masashi Ito and Kenji Kubara of Eisai Co. Ltd. for their valuable advice regarding the methods for FVIII measurement.

Author information

Authors and Affiliations

  1. Department of Pediatrics, University of Tsukuba Hospital, 2-1-1 Amakubo, Tsukuba, Ibaraki, 305-8576, Japan

    Yuni Yamaki, Takashi Fukushima & Hidetoshi Takada

  2. Department of Child Health, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan

    Takashi Fukushima, Ryo Sumazaki & Hidetoshi Takada

  3. Department of Pediatric Tumor, Saitama Medical University International Medical Center, Saitama, Japan

    Takashi Fukushima

  4. Biotechnology Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology, Ibaraki, Japan

    Naomi Yoshida & Mahito Nakanishi

  5. TOKIWA-Bio Inc., Ibaraki, Japan

    Naomi Yoshida & Mahito Nakanishi

  6. Graduate School of Comprehensive Human Sciences, University of Tsukuba, Ibaraki, Japan

    Ken Nishimura, Aya Fukuda & Koji Hisatake

  7. CellGenTech, Inc., Chiba, Japan

    Masayuki Aso

  8. Department of Molecular Pharmacology, Faculty of Medicine, University of Tsukuba, Ibaraki, Japan

    Tomoki Sakasai, Junko Kijima-Tanaka & Yoshihiro Miwa

  9. Ibaraki Children’s Hospital, Ibaraki, Japan

    Ryo Sumazaki

Authors
  1. Yuni Yamaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Takashi Fukushima
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Naomi Yoshida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ken Nishimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Aya Fukuda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Koji Hisatake
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Masayuki Aso
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tomoki Sakasai
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Junko Kijima-Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yoshihiro Miwa
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Mahito Nakanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Ryo Sumazaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Hidetoshi Takada
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

YY performed the research and wrote the manuscript; TF supervised all the experiments; NY and MN provided the SeVdp vectors and gave advice on their use in the study; KN, AF, and KH gave crucial advice on the experiments using 3T3-L1 and 3T3-F442A cells; MA gave crucial advice in consideration of the clinical application of this research; TS, JKT, and YM gave crucial advice for the iRFP experiments; HT assisted in the preparation of the manuscript; and RS designed the research study. All authors critically reviewed and approved the final manuscript.

Corresponding author

Correspondence to Yuni Yamaki.

Ethics declarations

Conflict of interest

MN is a founder and CTO of TOKIWA-Bio, Inc., and receives personal fees from TOKIWA-Bio, Inc. In addition, MN has a patent JP 4936482 issued, and a patent US 9145564 issued, relevant to this study. NY is an employee of TOKIWA-Bio, Inc. The other authors declare that there are no conflicts of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yamaki, Y., Fukushima, T., Yoshida, N. et al. Utilization of a novel Sendai virus vector in ex vivo gene therapy for hemophilia A. Int J Hematol 113, 493–499 (2021). https://doi.org/10.1007/s12185-020-03059-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12185-020-03059-6

Keywords

Search

Navigation