Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine

Robert A. Kaiser, Shennen A. Mao, Jaime Glorioso, Bruce Amiot, Clara T. Nicolas, Kari L. Allen, Zeji Du, Caitlin J. VanLith, Raymond D. Hickey, Scott L. Nyberg, Joseph B. Lillegard

Research output: Contribution to journalArticle

Abstract

Gene therapy is an ideal choice to cure many inborn errors of metabolism of the liver. Ex-vivo, lentiviral vectors have been used successfully in the treatment of many hematopoietic diseases in humans, as their use offers stable transgene expression due to the vector's ability to integrate into the host genome. This method demonstrates the application of ex vivo gene therapy of hepatocytes to a large animal model of hereditary tyrosinemia type I. This process consists of 1) isolation of primary hepatocytes from the autologous donor/recipient animal, 2) ex vivo gene delivery via hepatocyte transduction with a lentiviral vector, and 3) autologous transplant of corrected hepatocytes via portal vein injection. Success of the method generally relies upon efficient and sterile removal of the liver resection, careful handling of the excised specimen for isolation of viable hepatocytes sufficient for re-engrafting, high-percentage transduction of the isolated cells, and aseptic surgical procedures throughout to prevent infection. Technical failure at any of these steps will result in low yield of viable transduced hepatocytes for autologous transplant or infection of the donor/recipient animal. The pig model of human type 1 hereditary tyrosinemia (HT-1) chosen for this approach is uniquely amenable to such a method, as even a small percentage of engraftment of corrected cells will lead to repopulation of the liver with healthy cells based on a powerful selective advantage over native-diseased hepatocytes. Although this growth selection will not be true for all indications, this approach is a foundation for expansion into other indications and allows for manipulation of this environment to address additional diseases, both within the liver and beyond, while controlling for exposure to viral vector and opportunity for off-target toxicity and tumorigenicity.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number141
DOIs
StatePublished - Nov 4 2018
Externally publishedYes

Fingerprint

Gene therapy
Autologous Transplantation
Genetic Therapy
Hepatocytes
Swine
Liver
Tyrosinemias
Transplants
Animals
Autografts
Genes
Specimen Handling
Inborn Errors Metabolism
Portal Vein
Infection
Transgenes
Metabolism
Toxicity
Animal Models
Cells

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine. / Kaiser, Robert A.; Mao, Shennen A.; Glorioso, Jaime; Amiot, Bruce; Nicolas, Clara T.; Allen, Kari L.; Du, Zeji; VanLith, Caitlin J.; Hickey, Raymond D.; Nyberg, Scott L.; Lillegard, Joseph B.

In: Journal of visualized experiments : JoVE, No. 141, 04.11.2018.

Research output: Contribution to journalArticle

Kaiser, RA, Mao, SA, Glorioso, J, Amiot, B, Nicolas, CT, Allen, KL, Du, Z, VanLith, CJ, Hickey, RD, Nyberg, SL & Lillegard, JB 2018, 'Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine', Journal of visualized experiments : JoVE, no. 141. https://doi.org/10.3791/58399
Kaiser, Robert A. ; Mao, Shennen A. ; Glorioso, Jaime ; Amiot, Bruce ; Nicolas, Clara T. ; Allen, Kari L. ; Du, Zeji ; VanLith, Caitlin J. ; Hickey, Raymond D. ; Nyberg, Scott L. ; Lillegard, Joseph B. / Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine. In: Journal of visualized experiments : JoVE. 2018 ; No. 141.
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