A novel gene therapy strategy for elimination of prostate carcinoma cells from human bone marrow.
We report a novel means to purge bone marrow of a specific subset of prostate carcinoma cells based on transductional and genetic selectivity. Using both adenovirus-polylysine-DNA complexes and E1A/B-deleted replication-deficient adenoviruses, we have demonstrated a transductional preference of these vectors for the prostate carcinoma cell lines DU 145, LNCaP, and PC-3 over primary human bone marrow cells and the leukemia cell line KG-1. We have also shown a genetic selectivity of an anti-erbB-2 intracellular single-chain antibody (sFv) encoding adenovirus, Ad21, for the erbB-2-positive prostate carcinoma cell lines DU 145 and LNCaP. Delivery of Ad21 resulted in cytotoxicity to the DU 145 and LNCaP, but not PC-3, cell lines and reduced the clonogenic capacity of DU 145 cells cultured alone or mixed with various ratios of irradiated human bone marrow. Finally, quantitative, competitive reverse transcription polymerase chain reaction (QC-RT-PCR) analysis demonstrated that Ad21 could effectively reduce DU 145 and erbB-2-positive primary prostate tumor contamination in bone marrow cultures. Delivery of Ad21 had no effect on the ability of progenitor cells to form colonies. These results suggest that an anti-erbB-2 sFv-encoding adenoviral vector is efficacious for removal of erbB-2-positive prostate carcinoma cells from human bone marrow, and demonstrates a novel method for ex vivo genetic purge of malignant cells from bone marrow for autologous bone marrow transplantation (ABMT) therapy.
Kim
M
M
Gene Therapy Program, University of Alabama at Birmingham 35294, USA.
Wright
M
M
Deshane
J
J
Accavitti
M A
MA
Tilden
A
A
Saleh
M
M
Vaughan
W P
WP
Carabasi
M H
MH
Rogers
M D
MD
Hockett
R D
RD
Grizzle
W E
WE
Curiel
D T
DT
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000906:Antibodies; D001853:Bone Marrow / Q000502:physiology* / Q000821:virology; D001854:Bone Marrow Cells; D002277:Carcinoma / Q000235:genetics* / Q000473:pathology / Q000821:virology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000821:virology; D012333:RNA, Messenger / Q000096:biosynthesis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013234:Stem Cells / Q000502:physiology; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
D000906:Antibodies; D012333:RNA, Messenger; D018719:Erb-b2 Receptor Tyrosine Kinases