A Novel Gene Therapy Strategy for Elimination of Prostate Carcinoma Cells from Human Bone Marrow
- 20 January 1997
- journal article
- research article
- Published by Mary Ann Liebert Inc in Human Gene Therapy
- Vol. 8 (2), 157-170
- https://doi.org/10.1089/hum.1997.8.2-157
Abstract
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. Chemotherapy and immunotherapy have been utilized to purge human bone marrow of tumor cell populations to improve the prognostic outcome of patients undergoing autologous bone marrow transplantation (ABMT). In this report, we show that adenovirus-mediated delivery of an anti-erbB-2 single-chain intracellular antibody (sFv) purges human bone marrow of erbB-2-positive prostate carcinoma cells. Both adenovirus and adenovirus–polylysine-DNA (AdpL) vectors encoding the luciferase and β-galactosidase (β-Gal) reporter genes preferentially transduced prostate carcinoma cell lines, but did not transduce either primary bone marrow cells or a leukemia cell line. The adenovirus encoding the anti-erbB-2 sFv was cytotoxic to the erbB-2-positive prostate carcinoma cell lines, but not to hematopoietic progenitor cells, and was able to abrogate the clonogenicity of DU 145 prostate carcinoma cells mixed with irradiated bone marrow. Quantitative, competitive reverse transcription polymerase chain reaction (QC-RT-PCR) analysis showed that an erbB-2-positive prostate carcinoma cell line and primary prostate carcinoma disease were essentially eliminated from bone marrow cells when treated with the anti-erbB-2 sFv encoding adenovirus.Keywords
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