Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy: An in vivo study of adenovirus-mediated transfer of the human transforming growth factor β1 encoding gene

Kotaro Nishida, James D. Kang, Lars G. Gilbertson, Seong Hwan Moon, Jun Kyo Suh, Molly T. Vogt, Paul D. Robbins, Christopher H. Evans

Research output: Contribution to journalArticle

261 Citations (Scopus)

Abstract

Study Design. In vivo studies using a rabbit model to determine the biologic effects of direct, adenovirus-mediated transfer of a therapeutic gene to the intervertebral disc. Objectives. 1) To deliver an exogenous therapeutic gene to rabbit lumbar intervertebral discs in vivo, 2) to quantify the resulting amount of gene expression, and 3) to determine the effect on the biologic activity of the discs. Summary of Background Data. Although growth factors such as transforming growth factor β1 appear to have promising therapeutic properties, there currently is no practical method for sustained delivery of exogenous growth factors to the disc for the management of certain chronic types of disease (e.g., disc degeneration). A possible solution is to modify the disc cells genetically through gene transfer such that the cells manufacture the desired growth factors endogenously on a continuous basis. Methods. Saline, with or without virus, was injected directly into lumbar discs of 22 skeletally mature female New Zealand white rabbits. Group 1 (n = 11) received the adenovirus construct Ad/CMV-hTGFβ1 containing the therapeutic human transforming growth factor β1-encoding gene. Group 2 (n = 6) received adenovirus containing the luciferase marker gene. Group 3 (n = 5) received saline only. The rabbits were killed 1 week after injection. Immunohistochemical staining for human transforming growth factor β1 was performed on the disc tissues of one rabbit from Group 1. Nucleus pulposus tissues from the remaining rabbits were cultured in serumless medium. Bioassays were performed to determine human transforming growth factor β1 production and proteoglycan synthesis. Results. Discs injected with Ad/CMV-hTGFβ1 exhibited extensive and intense positive immunostaining for transforming growth factorβ1. The nucleus pulposus tissues from the discs injected with Ad/CMV-hTGFβ1 exhibited a 30-fold increase in active transforming growth factorβ1 production, and a 5-fold increase in total (active + latent) transforming growth factor β1 production over that from intact control discs (P < 0.05). Furthermore, these tissues exhibited a 100% increase in proteoglycan synthesis compared with intact control tissue, which was statistically significant (P < 0.05). Conclusions. The results of this study suggest that the intervertebral disc is an appropriate site for adenovirus-mediated transfer of exogenous genes and subsequent production of therapeutic growth factors. Gene therapy therefore may have useful applications for study of the basic science of the intervertebral disc and for clinical management of degenerative disc disease.

Original languageEnglish
Pages (from-to)2419-2425
Number of pages7
JournalSpine
Volume24
Issue number23
DOIs
Publication statusPublished - 1999 Dec 1

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Intervertebral Disc
Transforming Growth Factors
Adenoviridae
Genetic Therapy
Rabbits
Intercellular Signaling Peptides and Proteins
Genes
Proteoglycans
Therapeutics
Intervertebral Disc Degeneration
Growth
Luciferases
Biological Assay
Chronic Disease
Staining and Labeling
Viruses
Gene Expression
Injections

All Science Journal Classification (ASJC) codes

  • Orthopedics and Sports Medicine
  • Clinical Neurology

Cite this

Nishida, Kotaro ; Kang, James D. ; Gilbertson, Lars G. ; Moon, Seong Hwan ; Suh, Jun Kyo ; Vogt, Molly T. ; Robbins, Paul D. ; Evans, Christopher H. / Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy : An in vivo study of adenovirus-mediated transfer of the human transforming growth factor β1 encoding gene. In: Spine. 1999 ; Vol. 24, No. 23. pp. 2419-2425.
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title = "Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy: An in vivo study of adenovirus-mediated transfer of the human transforming growth factor β1 encoding gene",
abstract = "Study Design. In vivo studies using a rabbit model to determine the biologic effects of direct, adenovirus-mediated transfer of a therapeutic gene to the intervertebral disc. Objectives. 1) To deliver an exogenous therapeutic gene to rabbit lumbar intervertebral discs in vivo, 2) to quantify the resulting amount of gene expression, and 3) to determine the effect on the biologic activity of the discs. Summary of Background Data. Although growth factors such as transforming growth factor β1 appear to have promising therapeutic properties, there currently is no practical method for sustained delivery of exogenous growth factors to the disc for the management of certain chronic types of disease (e.g., disc degeneration). A possible solution is to modify the disc cells genetically through gene transfer such that the cells manufacture the desired growth factors endogenously on a continuous basis. Methods. Saline, with or without virus, was injected directly into lumbar discs of 22 skeletally mature female New Zealand white rabbits. Group 1 (n = 11) received the adenovirus construct Ad/CMV-hTGFβ1 containing the therapeutic human transforming growth factor β1-encoding gene. Group 2 (n = 6) received adenovirus containing the luciferase marker gene. Group 3 (n = 5) received saline only. The rabbits were killed 1 week after injection. Immunohistochemical staining for human transforming growth factor β1 was performed on the disc tissues of one rabbit from Group 1. Nucleus pulposus tissues from the remaining rabbits were cultured in serumless medium. Bioassays were performed to determine human transforming growth factor β1 production and proteoglycan synthesis. Results. Discs injected with Ad/CMV-hTGFβ1 exhibited extensive and intense positive immunostaining for transforming growth factorβ1. The nucleus pulposus tissues from the discs injected with Ad/CMV-hTGFβ1 exhibited a 30-fold increase in active transforming growth factorβ1 production, and a 5-fold increase in total (active + latent) transforming growth factor β1 production over that from intact control discs (P < 0.05). Furthermore, these tissues exhibited a 100{\%} increase in proteoglycan synthesis compared with intact control tissue, which was statistically significant (P < 0.05). Conclusions. The results of this study suggest that the intervertebral disc is an appropriate site for adenovirus-mediated transfer of exogenous genes and subsequent production of therapeutic growth factors. Gene therapy therefore may have useful applications for study of the basic science of the intervertebral disc and for clinical management of degenerative disc disease.",
author = "Kotaro Nishida and Kang, {James D.} and Gilbertson, {Lars G.} and Moon, {Seong Hwan} and Suh, {Jun Kyo} and Vogt, {Molly T.} and Robbins, {Paul D.} and Evans, {Christopher H.}",
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Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy : An in vivo study of adenovirus-mediated transfer of the human transforming growth factor β1 encoding gene. / Nishida, Kotaro; Kang, James D.; Gilbertson, Lars G.; Moon, Seong Hwan; Suh, Jun Kyo; Vogt, Molly T.; Robbins, Paul D.; Evans, Christopher H.

In: Spine, Vol. 24, No. 23, 01.12.1999, p. 2419-2425.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy

T2 - An in vivo study of adenovirus-mediated transfer of the human transforming growth factor β1 encoding gene

AU - Nishida, Kotaro

AU - Kang, James D.

AU - Gilbertson, Lars G.

AU - Moon, Seong Hwan

AU - Suh, Jun Kyo

AU - Vogt, Molly T.

AU - Robbins, Paul D.

AU - Evans, Christopher H.

PY - 1999/12/1

Y1 - 1999/12/1

N2 - Study Design. In vivo studies using a rabbit model to determine the biologic effects of direct, adenovirus-mediated transfer of a therapeutic gene to the intervertebral disc. Objectives. 1) To deliver an exogenous therapeutic gene to rabbit lumbar intervertebral discs in vivo, 2) to quantify the resulting amount of gene expression, and 3) to determine the effect on the biologic activity of the discs. Summary of Background Data. Although growth factors such as transforming growth factor β1 appear to have promising therapeutic properties, there currently is no practical method for sustained delivery of exogenous growth factors to the disc for the management of certain chronic types of disease (e.g., disc degeneration). A possible solution is to modify the disc cells genetically through gene transfer such that the cells manufacture the desired growth factors endogenously on a continuous basis. Methods. Saline, with or without virus, was injected directly into lumbar discs of 22 skeletally mature female New Zealand white rabbits. Group 1 (n = 11) received the adenovirus construct Ad/CMV-hTGFβ1 containing the therapeutic human transforming growth factor β1-encoding gene. Group 2 (n = 6) received adenovirus containing the luciferase marker gene. Group 3 (n = 5) received saline only. The rabbits were killed 1 week after injection. Immunohistochemical staining for human transforming growth factor β1 was performed on the disc tissues of one rabbit from Group 1. Nucleus pulposus tissues from the remaining rabbits were cultured in serumless medium. Bioassays were performed to determine human transforming growth factor β1 production and proteoglycan synthesis. Results. Discs injected with Ad/CMV-hTGFβ1 exhibited extensive and intense positive immunostaining for transforming growth factorβ1. The nucleus pulposus tissues from the discs injected with Ad/CMV-hTGFβ1 exhibited a 30-fold increase in active transforming growth factorβ1 production, and a 5-fold increase in total (active + latent) transforming growth factor β1 production over that from intact control discs (P < 0.05). Furthermore, these tissues exhibited a 100% increase in proteoglycan synthesis compared with intact control tissue, which was statistically significant (P < 0.05). Conclusions. The results of this study suggest that the intervertebral disc is an appropriate site for adenovirus-mediated transfer of exogenous genes and subsequent production of therapeutic growth factors. Gene therapy therefore may have useful applications for study of the basic science of the intervertebral disc and for clinical management of degenerative disc disease.

AB - Study Design. In vivo studies using a rabbit model to determine the biologic effects of direct, adenovirus-mediated transfer of a therapeutic gene to the intervertebral disc. Objectives. 1) To deliver an exogenous therapeutic gene to rabbit lumbar intervertebral discs in vivo, 2) to quantify the resulting amount of gene expression, and 3) to determine the effect on the biologic activity of the discs. Summary of Background Data. Although growth factors such as transforming growth factor β1 appear to have promising therapeutic properties, there currently is no practical method for sustained delivery of exogenous growth factors to the disc for the management of certain chronic types of disease (e.g., disc degeneration). A possible solution is to modify the disc cells genetically through gene transfer such that the cells manufacture the desired growth factors endogenously on a continuous basis. Methods. Saline, with or without virus, was injected directly into lumbar discs of 22 skeletally mature female New Zealand white rabbits. Group 1 (n = 11) received the adenovirus construct Ad/CMV-hTGFβ1 containing the therapeutic human transforming growth factor β1-encoding gene. Group 2 (n = 6) received adenovirus containing the luciferase marker gene. Group 3 (n = 5) received saline only. The rabbits were killed 1 week after injection. Immunohistochemical staining for human transforming growth factor β1 was performed on the disc tissues of one rabbit from Group 1. Nucleus pulposus tissues from the remaining rabbits were cultured in serumless medium. Bioassays were performed to determine human transforming growth factor β1 production and proteoglycan synthesis. Results. Discs injected with Ad/CMV-hTGFβ1 exhibited extensive and intense positive immunostaining for transforming growth factorβ1. The nucleus pulposus tissues from the discs injected with Ad/CMV-hTGFβ1 exhibited a 30-fold increase in active transforming growth factorβ1 production, and a 5-fold increase in total (active + latent) transforming growth factor β1 production over that from intact control discs (P < 0.05). Furthermore, these tissues exhibited a 100% increase in proteoglycan synthesis compared with intact control tissue, which was statistically significant (P < 0.05). Conclusions. The results of this study suggest that the intervertebral disc is an appropriate site for adenovirus-mediated transfer of exogenous genes and subsequent production of therapeutic growth factors. Gene therapy therefore may have useful applications for study of the basic science of the intervertebral disc and for clinical management of degenerative disc disease.

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