Transfer of genes to chondrocytic cells of the lumbar spine

Proposal for a treatment strategy of spinal disorders by local gene therapy

Peter Wehling, Klaus Peter Schulitz, Paul D. Robbins, Christopher H Evans, Julio A. Reinecke

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Study Design. In the current study, chondrocytic cells from bovine intervertebral end plates were cultivated in vitro end modified genetically. Objective. The authors intended to perform isolation and cultivation of cells from bovine end plates of the spine. They also intended to show, in principle, the feasibility of introducing exogenous genes into chondrocytic cells from bovine intervertebral end plates by way of retroviral vectors. Summary of Background Data. The involvement of cytokines in the destruction of articular cartilage is established. It appears possible that similar mechanisms may play a role in intervertebral disc degeneration and other spinal disorders. Conventional medication and surgery of intervertebral disc degeneration addresses neither the pathophysiology nor the chronicity of the disease. Therapeutic proteins carry great potential as locally produced drugs after transfer of their cognate genes to the sites of interest. Methods. Vertebral end plate tissue was obtained from bovine os coccygis. Chondrocytic cells were isolated and cultured in vitro. The bacterial β-galactosidase (LacZ) gene and, alternatively, the complementary DNA (DNA copy of the mRNA) of the human interleukin-1 receptor antagonist were introduced into the isolated cells by retrovirus mediated gene transfer. β-galactosidase activity was determined by staining, and interleukin-1 receptor antagonist protein was quantitated by enzyme-linked immunosorbent assay. Results. Isolation and cultivation of chondrocytic end plate cells is possible. Native cells continue to grow in culture for more than 2 months. Transfer of the β- galactosidase gene to cultured cells resulted in ~1% β-galactosidase positive cells. Transfer of the interleukin-1 receptor antagonist complementary DNA resulted in the production of 24 ng/ml/106 cells interleukin-1 receptor antagonist protein in 48 hours. Conclusions. The introduction of exogenous therapeutic genes into cells from the intervertebral end plate opens the possibility for s local gene-based treatment of intervertebral disc degeneration. This approach avoids some of the shortcomings of conventional drug- and surgery-based treatments and has the potential to be specific, effective, and appropriate to the chronicity of the disease.

Original languageEnglish (US)
Pages (from-to)1092-1097
Number of pages6
JournalSpine
Volume22
Issue number10
DOIs
StatePublished - May 15 1997
Externally publishedYes

Fingerprint

Genetic Therapy
Spine
Galactosidases
Genes
Intervertebral Disc Degeneration
Interleukin 1 Receptor Antagonist Protein
Interleukin-1 Receptors
Therapeutics
Complementary DNA
Lac Operon
Cell Separation
Articular Cartilage
Retroviridae
Pharmaceutical Preparations
Cultured Cells
Enzyme-Linked Immunosorbent Assay
Staining and Labeling
Cytokines
Messenger RNA
DNA

Keywords

  • back pain
  • chondrocytes
  • gene therapy
  • interleukin-1 receptor antagonist
  • intervertebral disc degeneration
  • retrovirus

ASJC Scopus subject areas

  • Physiology
  • Clinical Neurology
  • Orthopedics and Sports Medicine

Cite this

Transfer of genes to chondrocytic cells of the lumbar spine : Proposal for a treatment strategy of spinal disorders by local gene therapy. / Wehling, Peter; Schulitz, Klaus Peter; Robbins, Paul D.; Evans, Christopher H; Reinecke, Julio A.

In: Spine, Vol. 22, No. 10, 15.05.1997, p. 1092-1097.

Research output: Contribution to journalArticle

Wehling, Peter ; Schulitz, Klaus Peter ; Robbins, Paul D. ; Evans, Christopher H ; Reinecke, Julio A. / Transfer of genes to chondrocytic cells of the lumbar spine : Proposal for a treatment strategy of spinal disorders by local gene therapy. In: Spine. 1997 ; Vol. 22, No. 10. pp. 1092-1097.
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abstract = "Study Design. In the current study, chondrocytic cells from bovine intervertebral end plates were cultivated in vitro end modified genetically. Objective. The authors intended to perform isolation and cultivation of cells from bovine end plates of the spine. They also intended to show, in principle, the feasibility of introducing exogenous genes into chondrocytic cells from bovine intervertebral end plates by way of retroviral vectors. Summary of Background Data. The involvement of cytokines in the destruction of articular cartilage is established. It appears possible that similar mechanisms may play a role in intervertebral disc degeneration and other spinal disorders. Conventional medication and surgery of intervertebral disc degeneration addresses neither the pathophysiology nor the chronicity of the disease. Therapeutic proteins carry great potential as locally produced drugs after transfer of their cognate genes to the sites of interest. Methods. Vertebral end plate tissue was obtained from bovine os coccygis. Chondrocytic cells were isolated and cultured in vitro. The bacterial β-galactosidase (LacZ) gene and, alternatively, the complementary DNA (DNA copy of the mRNA) of the human interleukin-1 receptor antagonist were introduced into the isolated cells by retrovirus mediated gene transfer. β-galactosidase activity was determined by staining, and interleukin-1 receptor antagonist protein was quantitated by enzyme-linked immunosorbent assay. Results. Isolation and cultivation of chondrocytic end plate cells is possible. Native cells continue to grow in culture for more than 2 months. Transfer of the β- galactosidase gene to cultured cells resulted in ~1{\%} β-galactosidase positive cells. Transfer of the interleukin-1 receptor antagonist complementary DNA resulted in the production of 24 ng/ml/106 cells interleukin-1 receptor antagonist protein in 48 hours. Conclusions. The introduction of exogenous therapeutic genes into cells from the intervertebral end plate opens the possibility for s local gene-based treatment of intervertebral disc degeneration. This approach avoids some of the shortcomings of conventional drug- and surgery-based treatments and has the potential to be specific, effective, and appropriate to the chronicity of the disease.",
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AU - Reinecke, Julio A.

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KW - intervertebral disc degeneration

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