Transduction and apoptosis induction in the rat prostate, using adenovirus vectors.
Proapoptotic adenovirus vectors offer great promise for the treatment of cancer and nonmalignant conditions. Benign prostate hyperplasia (BPH) is a common nonmalignant enlargement of the prostate that involves epithelial, stromal, and smooth muscle components of the gland. We tested the hypothesis that an adenovirus vector expressing Fas ligand can be used to induce apoptosis in the prostate. We analyzed the efficiency of transduction and apoptosis induction in primary cultures of human prostate cells after adenovirus-mediated gene transfer. Efficient transduction was observed in primary prostate epithelial cells. Stromal and smooth muscle cells were more difficult to transduce, as no coxsackie-adenovirus receptor (CAR) expression was detectable on these cells. However, transduction was achieved in these cells when the multiplicity of infection was increased to 100 focal-forming units per cell, or when the vectors were delivered as calcium phosphate precipitates. Infection of all three primary prostate cell types with an adenovirus vector that expresses Fas ligand (AdFasL/G) resulted in rapid apoptosis. Direct injection of the rat prostate with an adenovirus vector carrying luciferase resulted in substantial luciferase expression. TUNEL analysis demonstrated that AdFasL/G administration induced low-level apoptosis in prostatic epithelial cells throughout the gland. As a first step toward enhancing the efficiency of prostate transduction in vivo, we tested an adenovirus vector that was engineered to have an expanded tropism. This vector, AdZ.F2K(pK7), was 10- to 500-fold more efficient than unmodified vectors in transducing prostate epithelial, smooth muscle, and stromal cells in culture. Moreover, AdZ.F2K(pK7) was more efficient than an unmodified vector at transducing the rat prostate in vivo, although the effect was dose dependent.
Kirkman
W
W
GenVec, Gaithersburg, MD 20878, USA.
Chen
P
P
Schroeder
R
R
Feneley
M R
MR
Rodriguez
R
R
Wickham
T J
TJ
King
C R
CR
Bruder
J T
JT
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D002130:Calcium Phosphates / Q000378:metabolism; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000378:metabolism; D053222:Fas Ligand Protein; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D006965:Hyperplasia / Q000628:therapy*; D020287:In Situ Nick-End Labeling; D008156:Luciferases / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D011467:Prostate / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000378:metabolism
D002130:Calcium Phosphates; C505661:FASLG protein, human; D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D008156:Luciferases; D011065:Poly(ADP-ribose) Polymerases; D001616:beta-Galactosidase; C485829:calcium phosphate, dibasic, anhydrous