Developing Adenoviral Vectors Encoding Therapeutic Genes Toxic to Host Cells: Comparing Binary and Single-inducible Vectors Expressing Truncated E2F-1

Overview

Journal Virology

Specialty Microbiology

Date 2009 Dec 17

PMID 20003994

Citations 6

Authors

Jorge G Gomez-Gutierrez

Xiao-Mei Rao

Aracely Garcia-Garcia

Hongying Hao

Kelly M McMasters

H Sam Zhou

Affiliations

Soon will be listed here.

Abstract

Adenoviral vectors are highly efficient at transferring genes into cells and are broadly used in cancer gene therapy. However, many therapeutic genes are toxic to vector host cells and thus inhibit vector production. The truncated form of E2F-1 (E2Ftr), which lacks the transactivation domain, can significantly induce cancer cell apoptosis, but is also toxic to HEK-293 cells and inhibits adenovirus replication. To overcome this, we have developed binary- and single-vector systems with a modified tetracycline-off inducible promoter to control E2Ftr expression. We compared several vectors and found that the structure of expression cassettes in vectors significantly affects E2Ftr expression. One construct expresses high levels of inducible E2Ftr and efficiently causes apoptotic cancer cell death by activation of caspase-3. The approach developed in this study may be applied in other viral vectors for encoding therapeutic genes that are toxic to their host cells and/or inhibit vector propagation.

Citing Articles

Adenovirus Lacking E1b Efficiently Induces Cytopathic Effect in HPV-16-Positive Murine Cancer Cells via Virus Replication and Apoptosis.

Martinez-Jaramillo E, Garza-Morales R, Wechman S, Montes de Oca-Luna R, Saucedo-Cardenas O, Shirwan H Cancer Invest. 2018; 36(1):19-27.

PMID: 29388837 PMC: 6088374. DOI: 10.1080/07357907.2018.1430812.

Enhanced cancer cell killing by truncated E2F-1 used in combination with oncolytic adenovirus.

Gomez-Gutierrez J, Rao X, Zhou H, McMasters K Virology. 2012; 433(2):538-47.

PMID: 23021422 PMC: 3494286. DOI: 10.1016/j.virol.2012.09.003.

E2F-1 lacking the transcriptional activity domain induces autophagy.

Garcia-Garcia A, Rodriguez-Rocha H, Tseng M, Montes de Oca-Luna R, Zhou H, McMasters K Cancer Biol Ther. 2012; 13(11):1091-101.

PMID: 22825328 PMC: 3462036. DOI: 10.4161/cbt.21143.

E2F-1- and E2Ftr-mediated apoptosis: the role of DREAM and HRK.

Hao H, Chen C, Rao X, Gomez-Gutierrez J, Zhou H, McMasters K J Cell Mol Med. 2011; 16(3):605-15.

PMID: 21564512 PMC: 3822935. DOI: 10.1111/j.1582-4934.2011.01338.x.

Constitutive Gs activation using a single-construct tetracycline-inducible expression system in embryonic stem cells and mice.

Hsiao E, Nguyen T, Ng J, Scott M, Chang W, Zahed H Stem Cell Res Ther. 2011; 2(2):11.

PMID: 21375737 PMC: 3226282. DOI: 10.1186/scrt52.

References

Fernandes-Alnemri T, Litwack G, Alnemri E . CPP32, a novel human apoptotic protein with homology to Caenorhabditis elegans cell death protein Ced-3 and mammalian interleukin-1 beta-converting enzyme. J Biol Chem. 1994; 269(49):30761-4. View

Bell L, OPrey J, Ryan K . DNA-binding independent cell death from a minimal proapoptotic region of E2F-1. Oncogene. 2006; 25(41):5656-63. DOI: 10.1038/sj.onc.1209580. View

Kovesdi I, Reichel R, Nevins J . E1A transcription induction: enhanced binding of a factor to upstream promoter sequences. Science. 1986; 231(4739):719-22. DOI: 10.1126/science.2935935. View

Hao H, Dong Y, Bowling M, Gomez-Gutierrez J, Zhou H, McMasters K . E2F-1 induces melanoma cell apoptosis via PUMA up-regulation and Bax translocation. BMC Cancer. 2007; 7:24. PMC: 1797184. DOI: 10.1186/1471-2407-7-24. View

Gomez-Gutierrez J, Souza V, Hao H, Montes de Oca-Luna R, Dong Y, Zhou H . Adenovirus-mediated gene transfer of FKHRL1 triple mutant efficiently induces apoptosis in melanoma cells. Cancer Biol Ther. 2006; 5(7):875-83. DOI: 10.4161/cbt.5.7.2911. View

Dong Y, Yang H, Elliott M, McMasters K . Adenovirus-mediated E2F-1 gene transfer sensitizes melanoma cells to apoptosis induced by topoisomerase II inhibitors. Cancer Res. 2002; 62(6):1776-83. View

Elliott M, Dong Y, Yang H, McMasters K . E2F-1 up-regulates c-Myc and p14(ARF) and induces apoptosis in colon cancer cells. Clin Cancer Res. 2001; 7(11):3590-7. View

Melillo R, Helin K, Lowy D, Schiller J . Positive and negative regulation of cell proliferation by E2F-1: influence of protein level and human papillomavirus oncoproteins. Mol Cell Biol. 1994; 14(12):8241-9. PMC: 359363. DOI: 10.1128/mcb.14.12.8241-8249.1994. View

Ermak G, Cancasci V, Davies K . Cytotoxic effect of doxycycline and its implications for tet-on gene expression systems. Anal Biochem. 2003; 318(1):152-4. DOI: 10.1016/s0003-2697(03)00166-0. View

10.

Vorburger S, Hetrakul N, Xia W, Wilson-Heiner M, Mirza N, Pollock R . Gene therapy with E2F-1 up-regulates the protein kinase PKR and inhibits growth of leiomyosarcoma in vivo. Mol Cancer Ther. 2005; 4(11):1710-6. DOI: 10.1158/1535-7163.MCT-05-0036. View

11.

Zhou H, Beaudet A . A new vector system with inducible E2a cell line for production of higher titer and safer adenoviral vectors. Virology. 2000; 275(2):348-57. DOI: 10.1006/viro.2000.0515. View

12.

Hsieh J, Fredersdorf S, Kouzarides T, Martin K, Lu X . E2F1-induced apoptosis requires DNA binding but not transactivation and is inhibited by the retinoblastoma protein through direct interaction. Genes Dev. 1997; 11(14):1840-52. DOI: 10.1101/gad.11.14.1840. View

13.

Dong Y, Phelps A, Yang H, Jamshidi-Parsian A, Chen C, Hao H . Induction of apoptosis signal-regulating Kinase 1 by E2F-1 may not be essential for E2F-1-mediated apoptosis in melanoma cells. Tumour Biol. 2007; 28(2):111-22. DOI: 10.1159/000099370. View

14.

Itoshima T, Fujiwara T, Waku T, Shao J, Kataoka M, Yarbrough W . Induction of apoptosis in human esophageal cancer cells by sequential transfer of the wild-type p53 and E2F-1 genes: involvement of p53 accumulation via ARF-mediated MDM2 down-regulation. Clin Cancer Res. 2000; 6(7):2851-9. View

15.

Fan J, Bertino J . Functional roles of E2F in cell cycle regulation. Oncogene. 1997; 14(10):1191-200. DOI: 10.1038/sj.onc.1200940. View

16.

Huang M, Hearing P . The adenovirus early region 4 open reading frame 6/7 protein regulates the DNA binding activity of the cellular transcription factor, E2F, through a direct complex. Genes Dev. 1989; 3(11):1699-710. DOI: 10.1101/gad.3.11.1699. View

17.

Dong Y, Yang H, Elliott M, Liu T, Stilwell A, Atienza Jr C . Adenovirus-mediated E2F-1 gene transfer efficiently induces apoptosis in melanoma cells. Cancer. 1999; 86(10):2021-33. DOI: 10.1002/(sici)1097-0142(19991115)86:10<2021::aid-cncr20>3.0.co;2-1. View

18.

Kovesdi I, Reichel R, Nevins J . Identification of a cellular transcription factor involved in E1A trans-activation. Cell. 1986; 45(2):219-28. DOI: 10.1016/0092-8674(86)90386-7. View

19.

Gossen M, Bujard H . Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A. 1992; 89(12):5547-51. PMC: 49329. DOI: 10.1073/pnas.89.12.5547. View

20.

Zheng X, Rao X, Gomez-Gutierrez J, Hao H, McMasters K, Zhou H . Adenovirus E1B55K region is required to enhance cyclin E expression for efficient viral DNA replication. J Virol. 2008; 82(7):3415-27. PMC: 2268468. DOI: 10.1128/JVI.01708-07. View