Gene therapy of murine solid tumors with T cells transduced with a retroviral vascular endothelial growth factor-immunotoxin target gene

Ni Jin, Wei Chen, Bruce R. Blazar, Sundaram Ramakrishnan, Daniel A. Vallera

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Solid tumor growth can be inhibited by targeting its neovasculature with vascular endothelial growth factor (VEGF)-toxin fusion proteins (FPs), but these agents have been limited by their inability to localize at the tumor site. In this study, we devised a gene therapy approach intended to deliver VEGF-toxin directly to tumor. Antigen-specific cytotoxic T lymphocytes (CTLs) served as vehicles to deliver a retroviral VEGF-toxin fusion protein to its specific leukemia cell target in vivo. A retroviral vector was constructed for gene therapy with VEGF positioned downstream of its 27-amino acid leader sequence, which promoted secretion of a catalytic immunotoxin containing either truncated diphtheria toxin or Pseudomonas exotoxin A. VEGF was chosen on the basis of the expression of VEGF receptor on endothelial cells in the tumor neovasculature. The VEGF FP was first expressed and secreted by mammalian NIH 3T3 cells. Intracellular expression of both VEGF and toxin was verified by immunofluorescence. In vitro, supernatants collected from transfected cells specifically inhibited the growth of VEGF receptor-expressing human umbilical vein endothelial cells (HUVECs), but not a control cell line. In vivo findings correlated with in vitro findings. A retroviral vector containing the target gene and a nerve growth factor receptor (NGFR) reporter gene was used to transiently transduce T15, a CD8+ CTL line that specifically recognizes C1498, a lethal C57BL/6 myeloid tumor. Transduced T15 cells injected intravenously significantly inhibited the growth of subcutaneous tumor, whereas nontransduced controls did not. Together, these data indicate that gene therapy of T cells with retrovirus containing a VEGF-immunotoxin target gene may be a valid means of inhibiting a broad range of solid tumors dependent on angiogenesis.

Original languageEnglish (US)
Pages (from-to)497-508
Number of pages12
JournalHuman Gene Therapy
Volume13
Issue number4
DOIs
StatePublished - Apr 6 2002
Externally publishedYes

Fingerprint

Immunotoxins
Genetic Therapy
Vascular Endothelial Growth Factor A
T-Lymphocytes
Genes
Neoplasms
Vascular Endothelial Growth Factor Receptor
Cytotoxic T-Lymphocytes
Growth
Diphtheria Toxin
Nerve Growth Factor Receptor
NIH 3T3 Cells
Proteins
Human Umbilical Vein Endothelial Cells
Retroviridae
Reporter Genes
Fluorescent Antibody Technique
Amino Acid Sequence
Leukemia
Endothelial Cells

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Gene therapy of murine solid tumors with T cells transduced with a retroviral vascular endothelial growth factor-immunotoxin target gene. / Jin, Ni; Chen, Wei; Blazar, Bruce R.; Ramakrishnan, Sundaram; Vallera, Daniel A.

In: Human Gene Therapy, Vol. 13, No. 4, 06.04.2002, p. 497-508.

Research output: Contribution to journalArticle

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