Gene therapy for prostate cancer: Toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters

E. Ebara, S. Shimura, Yasutomo Nasu, H. Kaku, H. Kumon, G. Yang, J. Wang, T. L. Timme, E. Aguilar-Cordova, T. C. Thompson

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Adenoviral vector delivery of the Herpes simplex virus thymidine kinase (HSV-tk) gene in combination with the prodrug ganciclovir (GCV) has been tested in phase I clinical trials for prostate cancer and found to exhibit a satisfactory toxicity profile. We have developed additional adenoviral vectors with differing promoters to optimize the expression profile and in the present study evaluate the potential systemic toxicity of these vectors. Four recombinant adenoviral vectors that express the HSV-tk gene were generated using three different promoters: CMV (leftward orientation); RSV (both rightward and leftward orientation); and the mouse caveolin-1 (cav-1) promoter (leftward orientation). Efficacy was determined in vitro by cytotoxicity assays in a mouse prostate cancer cell line, RM-9, and in vivo by treating orthotopic tumors. Potential toxicity was evaluated from liver histology and apoptotic cell counts and enzyme levels in the serum following intravenous adenoviral vector injection. Although there were differences in HSV-tk expression at the protein level among the four vectors there were no significant differences in in-vitro cytotoxicity studies with GCV or in vivo in tumor growth suppression of an orthotopic mouse prostate cancer model in GCV treated mice. Intravenous delivery of high doses of all adenoviral vectors lead to abnormalities in liver function as measured by specific serum markers and histological evaluation of liver tissue and increased levels of apoptosis in the liver. These abnormalities were most prevalent with the vector containing the CMV promoter and the rightward oriented RSV promoter. They were least prevalent in the vector regulated by the cav-1 promoter. Upregulation of specific chemokines, MIP-2 and MIP-1β was correlated with apoptotic counts. Our results demonstrate that comprehensive toxicological analysis of adenoviral vectors provides internally consistent information that can differentiate vectors with comparable efficacy based on toxicity. In these studies vectors with the cav-1 promoter-driven and leftward RSV-driven HSV-tk gene demonstrated minimal toxicities with cytotoxic effectiveness comparable to more toxic vectors. Our studies further suggest that promoter selection can influence the toxic effects of an adenoviral gene therapy vector.

Original languageEnglish
Pages (from-to)316-325
Number of pages10
JournalProstate Cancer and Prostatic Diseases
Volume5
Issue number4
DOIs
Publication statusPublished - 2002

Fingerprint

Thymidine Kinase
Simplexvirus
Caveolin 1
Genetic Therapy
Toxicology
Ganciclovir
Prostatic Neoplasms
Poisons
Liver
Chemokine CXCL2
Genes
Clinical Trials, Phase I
Prodrugs
Neoplasms
Histology
Up-Regulation
Cell Count
Biomarkers
Apoptosis
Cell Line

Keywords

  • Caveolin-1 promoter
  • Liver toxicity
  • Orthotopic tumor model
  • Prostate cancer
  • Suicide gene therapy

ASJC Scopus subject areas

  • Oncology
  • Urology

Cite this

Gene therapy for prostate cancer : Toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters. / Ebara, E.; Shimura, S.; Nasu, Yasutomo; Kaku, H.; Kumon, H.; Yang, G.; Wang, J.; Timme, T. L.; Aguilar-Cordova, E.; Thompson, T. C.

In: Prostate Cancer and Prostatic Diseases, Vol. 5, No. 4, 2002, p. 316-325.

Research output: Contribution to journalArticle

Ebara, E. ; Shimura, S. ; Nasu, Yasutomo ; Kaku, H. ; Kumon, H. ; Yang, G. ; Wang, J. ; Timme, T. L. ; Aguilar-Cordova, E. ; Thompson, T. C. / Gene therapy for prostate cancer : Toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters. In: Prostate Cancer and Prostatic Diseases. 2002 ; Vol. 5, No. 4. pp. 316-325.
@article{2c0ec6d5907149398f6f2532cd42657d,
title = "Gene therapy for prostate cancer: Toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters",
abstract = "Adenoviral vector delivery of the Herpes simplex virus thymidine kinase (HSV-tk) gene in combination with the prodrug ganciclovir (GCV) has been tested in phase I clinical trials for prostate cancer and found to exhibit a satisfactory toxicity profile. We have developed additional adenoviral vectors with differing promoters to optimize the expression profile and in the present study evaluate the potential systemic toxicity of these vectors. Four recombinant adenoviral vectors that express the HSV-tk gene were generated using three different promoters: CMV (leftward orientation); RSV (both rightward and leftward orientation); and the mouse caveolin-1 (cav-1) promoter (leftward orientation). Efficacy was determined in vitro by cytotoxicity assays in a mouse prostate cancer cell line, RM-9, and in vivo by treating orthotopic tumors. Potential toxicity was evaluated from liver histology and apoptotic cell counts and enzyme levels in the serum following intravenous adenoviral vector injection. Although there were differences in HSV-tk expression at the protein level among the four vectors there were no significant differences in in-vitro cytotoxicity studies with GCV or in vivo in tumor growth suppression of an orthotopic mouse prostate cancer model in GCV treated mice. Intravenous delivery of high doses of all adenoviral vectors lead to abnormalities in liver function as measured by specific serum markers and histological evaluation of liver tissue and increased levels of apoptosis in the liver. These abnormalities were most prevalent with the vector containing the CMV promoter and the rightward oriented RSV promoter. They were least prevalent in the vector regulated by the cav-1 promoter. Upregulation of specific chemokines, MIP-2 and MIP-1β was correlated with apoptotic counts. Our results demonstrate that comprehensive toxicological analysis of adenoviral vectors provides internally consistent information that can differentiate vectors with comparable efficacy based on toxicity. In these studies vectors with the cav-1 promoter-driven and leftward RSV-driven HSV-tk gene demonstrated minimal toxicities with cytotoxic effectiveness comparable to more toxic vectors. Our studies further suggest that promoter selection can influence the toxic effects of an adenoviral gene therapy vector.",
keywords = "Caveolin-1 promoter, Liver toxicity, Orthotopic tumor model, Prostate cancer, Suicide gene therapy",
author = "E. Ebara and S. Shimura and Yasutomo Nasu and H. Kaku and H. Kumon and G. Yang and J. Wang and Timme, {T. L.} and E. Aguilar-Cordova and Thompson, {T. C.}",
year = "2002",
doi = "10.1038/sj.pcan.4500610",
language = "English",
volume = "5",
pages = "316--325",
journal = "Prostate Cancer and Prostatic Diseases",
issn = "1365-7852",
publisher = "Nature Publishing Group",
number = "4",

}

TY - JOUR

T1 - Gene therapy for prostate cancer

T2 - Toxicological profile of four HSV-tk transducing adenoviral vectors regulated by different promoters

AU - Ebara, E.

AU - Shimura, S.

AU - Nasu, Yasutomo

AU - Kaku, H.

AU - Kumon, H.

AU - Yang, G.

AU - Wang, J.

AU - Timme, T. L.

AU - Aguilar-Cordova, E.

AU - Thompson, T. C.

PY - 2002

Y1 - 2002

N2 - Adenoviral vector delivery of the Herpes simplex virus thymidine kinase (HSV-tk) gene in combination with the prodrug ganciclovir (GCV) has been tested in phase I clinical trials for prostate cancer and found to exhibit a satisfactory toxicity profile. We have developed additional adenoviral vectors with differing promoters to optimize the expression profile and in the present study evaluate the potential systemic toxicity of these vectors. Four recombinant adenoviral vectors that express the HSV-tk gene were generated using three different promoters: CMV (leftward orientation); RSV (both rightward and leftward orientation); and the mouse caveolin-1 (cav-1) promoter (leftward orientation). Efficacy was determined in vitro by cytotoxicity assays in a mouse prostate cancer cell line, RM-9, and in vivo by treating orthotopic tumors. Potential toxicity was evaluated from liver histology and apoptotic cell counts and enzyme levels in the serum following intravenous adenoviral vector injection. Although there were differences in HSV-tk expression at the protein level among the four vectors there were no significant differences in in-vitro cytotoxicity studies with GCV or in vivo in tumor growth suppression of an orthotopic mouse prostate cancer model in GCV treated mice. Intravenous delivery of high doses of all adenoviral vectors lead to abnormalities in liver function as measured by specific serum markers and histological evaluation of liver tissue and increased levels of apoptosis in the liver. These abnormalities were most prevalent with the vector containing the CMV promoter and the rightward oriented RSV promoter. They were least prevalent in the vector regulated by the cav-1 promoter. Upregulation of specific chemokines, MIP-2 and MIP-1β was correlated with apoptotic counts. Our results demonstrate that comprehensive toxicological analysis of adenoviral vectors provides internally consistent information that can differentiate vectors with comparable efficacy based on toxicity. In these studies vectors with the cav-1 promoter-driven and leftward RSV-driven HSV-tk gene demonstrated minimal toxicities with cytotoxic effectiveness comparable to more toxic vectors. Our studies further suggest that promoter selection can influence the toxic effects of an adenoviral gene therapy vector.

AB - Adenoviral vector delivery of the Herpes simplex virus thymidine kinase (HSV-tk) gene in combination with the prodrug ganciclovir (GCV) has been tested in phase I clinical trials for prostate cancer and found to exhibit a satisfactory toxicity profile. We have developed additional adenoviral vectors with differing promoters to optimize the expression profile and in the present study evaluate the potential systemic toxicity of these vectors. Four recombinant adenoviral vectors that express the HSV-tk gene were generated using three different promoters: CMV (leftward orientation); RSV (both rightward and leftward orientation); and the mouse caveolin-1 (cav-1) promoter (leftward orientation). Efficacy was determined in vitro by cytotoxicity assays in a mouse prostate cancer cell line, RM-9, and in vivo by treating orthotopic tumors. Potential toxicity was evaluated from liver histology and apoptotic cell counts and enzyme levels in the serum following intravenous adenoviral vector injection. Although there were differences in HSV-tk expression at the protein level among the four vectors there were no significant differences in in-vitro cytotoxicity studies with GCV or in vivo in tumor growth suppression of an orthotopic mouse prostate cancer model in GCV treated mice. Intravenous delivery of high doses of all adenoviral vectors lead to abnormalities in liver function as measured by specific serum markers and histological evaluation of liver tissue and increased levels of apoptosis in the liver. These abnormalities were most prevalent with the vector containing the CMV promoter and the rightward oriented RSV promoter. They were least prevalent in the vector regulated by the cav-1 promoter. Upregulation of specific chemokines, MIP-2 and MIP-1β was correlated with apoptotic counts. Our results demonstrate that comprehensive toxicological analysis of adenoviral vectors provides internally consistent information that can differentiate vectors with comparable efficacy based on toxicity. In these studies vectors with the cav-1 promoter-driven and leftward RSV-driven HSV-tk gene demonstrated minimal toxicities with cytotoxic effectiveness comparable to more toxic vectors. Our studies further suggest that promoter selection can influence the toxic effects of an adenoviral gene therapy vector.

KW - Caveolin-1 promoter

KW - Liver toxicity

KW - Orthotopic tumor model

KW - Prostate cancer

KW - Suicide gene therapy

UR - http://www.scopus.com/inward/record.url?scp=0036968378&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036968378&partnerID=8YFLogxK

U2 - 10.1038/sj.pcan.4500610

DO - 10.1038/sj.pcan.4500610

M3 - Article

C2 - 12627218

AN - SCOPUS:0036968378

VL - 5

SP - 316

EP - 325

JO - Prostate Cancer and Prostatic Diseases

JF - Prostate Cancer and Prostatic Diseases

SN - 1365-7852

IS - 4

ER -