Therapeutic levels of human Factor VIII in mice implanted with encapsulated cells

Potential for gene therapy of haemophilia A

Carmen García-Martín, Marinee K L Chuah, An Van Damme, Kelly E. Robinson, Beatrijs Vanzieleghem, Jean Marie Saint-Remy, Dominique Gallardo, Frederick A. Ofosu, Thierry Vandendriessche, Gonzalo Hortelano

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein has been evaluated. The microcapsules are implanted intraperitoneally. In order to prevent cell immune rejection, cells are enclosed in non-antigenic biocompatible alginate microcapsules prior to their implantation into mice. It has been shown that encapsulated myoblasts can deliver therapeutic levels of Factor IX (FIX) in mice. The delivery of human Factor VIII (hFVIII) in mice using microcapsules was evaluated in this study. Methods: Mouse C2C12 myoblasts and canine MDCK epithelial kidney cells were transduced with MFG-FVIII (B-domain deleted) vector. Selected recombinant clones were enclosed in alginate microcapsules. Encapsulated recombinant clones were subsequently implanted intraperitoneally into C57BL/6 and immunodeficient SCID mice. Results: Plasma of mice receiving C2C12 and encapsulated MDCK cells had transient therapeutic levels of FVIII in immunocompetent C57BL/6 mice (up to 20% and 7% of physiological levels, respectively). In addition, FVIII delivery in SCID mice was also transient, suggesting that a non-immune mechanism must have contributed to the decline of hFVIII in plasma. Quantitative RT-PCR analysis confirmed directly that the decline of hFVIII is due to a reduction in steady-state hFVIII mRNA, consistent with transcriptional repression. Furthermore, encapsulated cells retrieved from implanted mice were viable, but secreted FVIII ex vivo at three-fold lower levels than the pre-implantation levels. In addition, antibodies to hFVIII were detected in immunocompetent C57BL/6 mice. Conclusions: Implantable microcapsules can deliver therapeutic levels of FVIII in mice, suggesting the potential of this gene therapy approach for haemophilia A. The findings suggest vector down-regulation in vivo.

Original languageEnglish
Pages (from-to)215-223
Number of pages9
JournalJournal of Gene Medicine
Volume4
Issue number2
DOIs
Publication statusPublished - Mar 2002
Externally publishedYes

Fingerprint

Hemophilia A
Genetic Therapy
Capsules
Madin Darby Canine Kidney Cells
SCID Mice
Myoblasts
Inbred C57BL Mouse
Therapeutics
Clone Cells
Gene Transfer Techniques
Factor IX
human F8 protein
Canidae
Down-Regulation
Epithelial Cells
Kidney
Polymerase Chain Reaction
Messenger RNA
Antibodies
Proteins

Keywords

  • Alginate
  • Factor VIII
  • Haemophilia A
  • Microcapsules
  • Myoblasts

ASJC Scopus subject areas

  • Genetics

Cite this

Therapeutic levels of human Factor VIII in mice implanted with encapsulated cells : Potential for gene therapy of haemophilia A. / García-Martín, Carmen; Chuah, Marinee K L; Van Damme, An; Robinson, Kelly E.; Vanzieleghem, Beatrijs; Saint-Remy, Jean Marie; Gallardo, Dominique; Ofosu, Frederick A.; Vandendriessche, Thierry; Hortelano, Gonzalo.

In: Journal of Gene Medicine, Vol. 4, No. 2, 03.2002, p. 215-223.

Research output: Contribution to journalArticle

García-Martín, C, Chuah, MKL, Van Damme, A, Robinson, KE, Vanzieleghem, B, Saint-Remy, JM, Gallardo, D, Ofosu, FA, Vandendriessche, T & Hortelano, G 2002, 'Therapeutic levels of human Factor VIII in mice implanted with encapsulated cells: Potential for gene therapy of haemophilia A', Journal of Gene Medicine, vol. 4, no. 2, pp. 215-223. https://doi.org/10.1002/jgm.248
García-Martín, Carmen ; Chuah, Marinee K L ; Van Damme, An ; Robinson, Kelly E. ; Vanzieleghem, Beatrijs ; Saint-Remy, Jean Marie ; Gallardo, Dominique ; Ofosu, Frederick A. ; Vandendriessche, Thierry ; Hortelano, Gonzalo. / Therapeutic levels of human Factor VIII in mice implanted with encapsulated cells : Potential for gene therapy of haemophilia A. In: Journal of Gene Medicine. 2002 ; Vol. 4, No. 2. pp. 215-223.
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abstract = "Background: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein has been evaluated. The microcapsules are implanted intraperitoneally. In order to prevent cell immune rejection, cells are enclosed in non-antigenic biocompatible alginate microcapsules prior to their implantation into mice. It has been shown that encapsulated myoblasts can deliver therapeutic levels of Factor IX (FIX) in mice. The delivery of human Factor VIII (hFVIII) in mice using microcapsules was evaluated in this study. Methods: Mouse C2C12 myoblasts and canine MDCK epithelial kidney cells were transduced with MFG-FVIII (B-domain deleted) vector. Selected recombinant clones were enclosed in alginate microcapsules. Encapsulated recombinant clones were subsequently implanted intraperitoneally into C57BL/6 and immunodeficient SCID mice. Results: Plasma of mice receiving C2C12 and encapsulated MDCK cells had transient therapeutic levels of FVIII in immunocompetent C57BL/6 mice (up to 20{\%} and 7{\%} of physiological levels, respectively). In addition, FVIII delivery in SCID mice was also transient, suggesting that a non-immune mechanism must have contributed to the decline of hFVIII in plasma. Quantitative RT-PCR analysis confirmed directly that the decline of hFVIII is due to a reduction in steady-state hFVIII mRNA, consistent with transcriptional repression. Furthermore, encapsulated cells retrieved from implanted mice were viable, but secreted FVIII ex vivo at three-fold lower levels than the pre-implantation levels. In addition, antibodies to hFVIII were detected in immunocompetent C57BL/6 mice. Conclusions: Implantable microcapsules can deliver therapeutic levels of FVIII in mice, suggesting the potential of this gene therapy approach for haemophilia A. The findings suggest vector down-regulation in vivo.",
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AU - García-Martín, Carmen

AU - Chuah, Marinee K L

AU - Van Damme, An

AU - Robinson, Kelly E.

AU - Vanzieleghem, Beatrijs

AU - Saint-Remy, Jean Marie

AU - Gallardo, Dominique

AU - Ofosu, Frederick A.

AU - Vandendriessche, Thierry

AU - Hortelano, Gonzalo

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N2 - Background: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein has been evaluated. The microcapsules are implanted intraperitoneally. In order to prevent cell immune rejection, cells are enclosed in non-antigenic biocompatible alginate microcapsules prior to their implantation into mice. It has been shown that encapsulated myoblasts can deliver therapeutic levels of Factor IX (FIX) in mice. The delivery of human Factor VIII (hFVIII) in mice using microcapsules was evaluated in this study. Methods: Mouse C2C12 myoblasts and canine MDCK epithelial kidney cells were transduced with MFG-FVIII (B-domain deleted) vector. Selected recombinant clones were enclosed in alginate microcapsules. Encapsulated recombinant clones were subsequently implanted intraperitoneally into C57BL/6 and immunodeficient SCID mice. Results: Plasma of mice receiving C2C12 and encapsulated MDCK cells had transient therapeutic levels of FVIII in immunocompetent C57BL/6 mice (up to 20% and 7% of physiological levels, respectively). In addition, FVIII delivery in SCID mice was also transient, suggesting that a non-immune mechanism must have contributed to the decline of hFVIII in plasma. Quantitative RT-PCR analysis confirmed directly that the decline of hFVIII is due to a reduction in steady-state hFVIII mRNA, consistent with transcriptional repression. Furthermore, encapsulated cells retrieved from implanted mice were viable, but secreted FVIII ex vivo at three-fold lower levels than the pre-implantation levels. In addition, antibodies to hFVIII were detected in immunocompetent C57BL/6 mice. Conclusions: Implantable microcapsules can deliver therapeutic levels of FVIII in mice, suggesting the potential of this gene therapy approach for haemophilia A. The findings suggest vector down-regulation in vivo.

AB - Background: A gene therapy delivery system based on microcapsules enclosing recombinant cells engineered to secrete a therapeutic protein has been evaluated. The microcapsules are implanted intraperitoneally. In order to prevent cell immune rejection, cells are enclosed in non-antigenic biocompatible alginate microcapsules prior to their implantation into mice. It has been shown that encapsulated myoblasts can deliver therapeutic levels of Factor IX (FIX) in mice. The delivery of human Factor VIII (hFVIII) in mice using microcapsules was evaluated in this study. Methods: Mouse C2C12 myoblasts and canine MDCK epithelial kidney cells were transduced with MFG-FVIII (B-domain deleted) vector. Selected recombinant clones were enclosed in alginate microcapsules. Encapsulated recombinant clones were subsequently implanted intraperitoneally into C57BL/6 and immunodeficient SCID mice. Results: Plasma of mice receiving C2C12 and encapsulated MDCK cells had transient therapeutic levels of FVIII in immunocompetent C57BL/6 mice (up to 20% and 7% of physiological levels, respectively). In addition, FVIII delivery in SCID mice was also transient, suggesting that a non-immune mechanism must have contributed to the decline of hFVIII in plasma. Quantitative RT-PCR analysis confirmed directly that the decline of hFVIII is due to a reduction in steady-state hFVIII mRNA, consistent with transcriptional repression. Furthermore, encapsulated cells retrieved from implanted mice were viable, but secreted FVIII ex vivo at three-fold lower levels than the pre-implantation levels. In addition, antibodies to hFVIII were detected in immunocompetent C57BL/6 mice. Conclusions: Implantable microcapsules can deliver therapeutic levels of FVIII in mice, suggesting the potential of this gene therapy approach for haemophilia A. The findings suggest vector down-regulation in vivo.

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