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PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types

Cancer Gene Therapy volume 16, pages 298–309 (2009)Cite this article

Abstract

Development of antineoplastic gene therapies is impaired by a paucity of transcription control elements with efficient, cancer cell-specific activity. We investigated the utility of promoter (AChP) and 5′-distal enhancer (ACE66) elements from the platelet-derived growth factor-A (PDGF-A) gene, which are hyperactive in many human cancers. Efficacy of these elements was tested in multiple tumor cell lines, both in cell culture and as tumor explants in athymic nude mice. Plasmid and viral vectors were constructed with the AChP promoter alone or in fusion with three copies of the ACE66 enhancer for expression of the prototype suicide gene, thymidine kinase (TK). ACE/AChP and AChP cassettes elicited ganciclovir (GCV)-induced cytotoxicity in multiple tumor cell lines. The ACE enhancer element also exhibited synergism with placental and liver-specific promoter elements. An adenovirus containing the AChP-TK cassette produced striking increases in GCV sensitivity in cultured tumor cell lines, as well as GCV-induced regression of U87 MG glioblastoma explants in vivo. TK expression was distributed throughout tumors receiving the therapeutic virus, whereas TdT-mediated dUTP nick end labeling (TUNEL) analysis revealed numerous regions undergoing apoptosis. Vascularization and reticulin fiber networks were less pronounced in virus-GCV-treated tumors, suggesting that both primary and stromal cell types may have been targeted. These studies provide proof-of-principle for utility of the PDGF-A promoter and ACE66 enhancer in antineoplastic gene therapy for a diverse group of human cancers.

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References

  1. Silver BJ . Platelet-derived growth factor in human malignancy. Biofactors 1992; 86: 217–227.

    Google Scholar 

  2. Heldin C-H, Westermark B . Mechanism of action and in vivo role of platelet-derived growth factor. Physiol Rev 1999; 79: 1283–1316.

    Article  CAS  Google Scholar 

  3. Sariban E, Sitaras NM, Antoniades HN, Kufe DW, Pantazis P . Expression of platelet-derived growth factor (PDGF)-related transcripts and synthesis of biologically active PDGF-like proteins by human malignant epithelial cell lines. J Clin Invest 1988; 82: 1157–1164.

    Article  CAS  Google Scholar 

  4. Fudge K, Bostwick DG, Stearns ME . Platelet-derived growth factor A and B chains and the α and β receptors in prostatic intraepithelial neoplasia. Prostate 1996; 29: 282–286.

    Article  CAS  Google Scholar 

  5. Beauchamp RD, Lyons RM, Yang EY, Coffey Jr RJ, Moses HL . Expression of and response to growth regulatory peptides by two human pancreatic carcinoma cell lines. Pancreas 1990; 5: 369–380.

    Article  CAS  Google Scholar 

  6. Cantley L, Auger KR, Carpenter C, Duckworth B, Graziani A, Kapeller R et al. Oncogenes and signal transduction. Cell 1991; 64: 281–302.

    Article  CAS  Google Scholar 

  7. Pietras K, Sjoblom T, Rubin K, Heldin CH, Ostman A . PDGF receptors as cancer drug targets. Cancer Cell 2003; 3: 439–443.

    Article  CAS  Google Scholar 

  8. Bonthron DT, Morton CC, Orkin SH, Collins T . Platelet-derived growth factor A-chain: gene structure, chromosomal location, and basis for alternative mRNA splicing. Proc Natl Acad Sci USA 1988; 85: 1492–1496.

    Article  CAS  Google Scholar 

  9. Kaetzel Jr DM, Maul RS, Liu B, Bonthron D, Fenstermaker RA, Coyne DW . Platelet-derived growth factor A-chain gene transcription is mediated by positive and negative regulatory regions in the promoter. Biochem J 1994; 301: 321–327.

    Article  CAS  Google Scholar 

  10. Zhang Q, Pedigo N, Shenoy S, Khalili K, Kaetzel DM . Purα activates PDGF-A gene transcription via interactions with a G-rich, single-stranded region of the promoter. Gene 2005; 348C: 25–32.

    Google Scholar 

  11. Gashler AL, Bonthron DT, Madden SL, Rauscher III FJ, Collins T, Sukhatme VP . Human platelet-derived growth factor A-chain is transcriptionally repressed by the Wilms tumor suppressor WT1. Proc Natl Acad Sci USA 1992; 89: 10984–10988.

    Article  CAS  Google Scholar 

  12. Wang Z-Y, Madden SL, Deuel TF, Rauscher III FJ . The Wilms’ tumor gene product, WT1, represses transcription of the platelet-derived growth factor A-chain gene. J Biol Chem 1992; 267: 21999–22002.

    CAS  PubMed  Google Scholar 

  13. Khachigian LM, Santiago FS, Rafty LA, Chan OL, Delbridge GJ, Bobik A et al. GC factor 2 represses platelet-derived growth factor A-chain gene transcription and is itself induced by arterial injury. Circ Res 1999; 84: 1258–1267.

    Article  CAS  Google Scholar 

  14. Rafty LA, Santiago FS, Khachigian LM . NF1/X represses PDGF A-chain transcription by interacting with Sp1 and antagonizing Sp1 occupancy of the promoter. EMBO J 2002; 21: 334–343.

    Article  CAS  Google Scholar 

  15. Maul RS, Zhang HX, Reid JD, Pedigo NG, Kaetzel DM . Identification of a cell type-specific enhancer in the distal 5'-region of the platelet-derived growth factor A-chain gene. J Biol Chem 1998; 273: 33239–33246.

    Article  CAS  Google Scholar 

  16. Pedigo N, Zhang H, Koszewski NJ, Kaetzel DM . A 5′-distal element mediates vitamin D-inducibility of PDGF-A gene transcription. Growth Factors 2003; 21: 151–160.

    Article  CAS  Google Scholar 

  17. Pedigo NG, Mishra A, Zhang H-X, McCorkle JR, Ormerod AK, Kaetzel DM . Retinoic acid-inducibility of the human PDGF-A gene is mediated by 5′-distal DNA motifs that overlap with basal enhancer and vitamin D response elements. Gene Expr 2007; 14: 1–12.

    Article  CAS  Google Scholar 

  18. Pedigo NG, Zhang H, Bruno ECM, Kaetzel CS, Dugan AR, Shanehsaz P et al. A 5′-distal enhanceosome in the PDGF-A gene is activated in choriocarcinoma cells via ligand-independent binding of vitamin D receptor and constitutive jun kinase signaling. Oncogene 2005; 24: 2654–2666.

    Article  CAS  Google Scholar 

  19. McKnight SL . The nucleotide sequence and transcript map of the herpes simplex virus thymidine kinase gene. Nucleic Acids Res 1980; 8: 5949–5964.

    Article  CAS  Google Scholar 

  20. Heckert LL, Schultz K, Nilson JH . Different composite regulatory elements direct expression of the human alpha subunit gene to pituitary and placenta. J Biol Chem 1995; 270: 26497–26504.

    Article  CAS  Google Scholar 

  21. Huang MC, Li KK, Spear BT . The mouse alpha-fetoprotein promoter is repressed in HepG2 hepatoma cells by hepatocyte nuclear factor-3 (FOXA). DNA Cell Biol 2002; 21: 561–569.

    Article  CAS  Google Scholar 

  22. Long L, Spear BT . FoxA proteins regulate H19 endoderm enhancer E1 and exhibit developmental changes in enhancer binding in vivo. Mol Cell Biol 2004; 24: 9601–9609.

    Article  CAS  Google Scholar 

  23. Kanegae Y, Makimura M, Saito I . A simple and efficient method for purification of infectious recombinant adenovirus. Jpn J Med Sci Biol 1994; 47: 157–166.

    Article  CAS  Google Scholar 

  24. Gonzalez M, Ruggiero FP, Chang Q, Shi YJ, Rich MM, Kraner S et al. Disruption of Trkb-mediated signaling induces disassembly of postsynaptic receptor clusters at neuromuscular junctions. Neuron 1999; 24: 567–583.

    Article  CAS  Google Scholar 

  25. De Lean A, Munson PJ, Rodbard D . Simultaneous analysis of sigmoidal curves: application to bioassay, radioligand assay, and physiological dose-response curves. Am J Physiol 1978; 235: E97–E102.

    Article  CAS  Google Scholar 

  26. Kaetzel DM . Transcription of the platelet-derived growth factor A-chain gene. Cytokine Growth Factor Rev 2003; 14: 427–446.

    Article  CAS  Google Scholar 

  27. Bokar JA, Keri RA, Farmerie TA, Fenstermaker RA, Andersen B, Hamernik DL et al. Expression of the glycoprotein hormone alpha-subunit gene in the placenta requires a functional cyclic AMP response element, whereas a different cis-acting element mediates pituitary-specific expression. Mol Cell Biol 1989; 9: 5113–5122.

    Article  CAS  Google Scholar 

  28. Spear BT, Tilghman SM . Role of alpha-fetoprotein regulatory elements in transcriptional activation in transient heterokaryons. Mol Cell Biol 1990; 10: 5047–5054.

    Article  CAS  Google Scholar 

  29. Yoo-Warren H, Pachnis V, Ingram RS, Tilghman SM . Two regulatory domains flank the mouse H19 gene. Mol Cell Biol 1988; 8: 4707–4715.

    Article  CAS  Google Scholar 

  30. Betsholtz C, Johnsson A, Heldin C-H, Westermark B, Lind P, Urdea MS et al. cDNA sequence and chromosomal localization of human platelet-derived growth factor A-chain and its expression in tumour cell lines. Nature 1986; 320: 695–699.

    Article  CAS  Google Scholar 

  31. Lindner V, Reidy MA . Platelet-derived growth factor ligand and receptor expression by large vessel endothelium in vivo. Am J Pathol 1995; 146: 1488–1497.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Sjolund M, Hedin U, Sejersen T, Heldin CH, Thyberg J . Arterial smooth muscle cells express platelet-derived growth factor (PDGF) A chain mRNA, secrete a PDGF-like mitogen, and bind exogenous PDGF in a phenotype- and growth state-dependent manner. J Cell Biol 1988; 106: 403–413.

    Article  CAS  Google Scholar 

  33. Majesky MW, Benditt EP, Schwartz SM . Expression and developmental control of platelet-derived growth factor A-chain and B-chain/Sis genes in rat aortic smooth muscle cells. Proc Natl Acad Sci USA 1988; 85: 1524–1528.

    Article  CAS  Google Scholar 

  34. Anan K, Morisaki T, Katano M, Ikubo A, Kitsuki H, Uchiyama A et al. Vascular endothelial growth factor and platelet-derived growth factor are potential angiogenic and metastatic factors in human breast cancer. Surgery 1996; 119: 333–339.

    Article  CAS  Google Scholar 

  35. Lippman ME, Dickson RB, Gelmann EP . Autocrine and paracrine growth regulation of human breast cancer. J Cell Biochem 1987; 35: 1–16.

    Article  CAS  Google Scholar 

  36. Peres R, Betsholtz C, Westermark B, Heldin CH . Frequent expression of growth factors for mesenchymal cells in human mammary carcinoma cell lines. Cancer Res 1987; 47: 3425–3429.

    CAS  PubMed  Google Scholar 

  37. Kalthoff H, Roeder C, Humburg I, Thiele HG, Greten H, Schmiegel W . Modulation of platelet-derived growth factor A- and B-chain/c-sis mRNA by tumor necrosis factor and other agents in adenocarcinoma cells. Oncogene 1991; 6: 1015–1021.

    CAS  PubMed  Google Scholar 

  38. Westermark B, Johnsson A, Paulsson Y, Betsholtz C, Heldin CH, Herlyn M et al. Human melanoma cell lines of primary and metastatic origin express the genes encoding the chains of platelet-derived growth factor (PDGF) and produce a PDGF-like growth factor. Proc Natl Acad Sci USA 1986; 83: 7197–7200.

    Article  CAS  Google Scholar 

  39. Shih IM, Herlyn M . Autocrine and paracrine roles for growth factors in melanoma. In Vivo 1994; 8: 113–123.

    CAS  PubMed  Google Scholar 

  40. Leveen P, Claesson-Welsh L, Heldin C-H, Westermark B, Betsholtz C . Expression of mRNAs for platelet-derived growth factor and its receptors in human sarcoma cell lines. Int J Sci 1990; 46: 1066–1070.

    CAS  Google Scholar 

  41. George D . Targeting PDGF receptors in cancer—rationales and proof of concept clinical trials. Adv Exp Med Biol 2003; 532: 141–151.

    Article  CAS  Google Scholar 

  42. Maher EA, Furnari FB, Bachoo RM, Rowitch DH, Louis DN, Cavenee WK et al. Malignant glioma: genetics and biology of a grave matter. Genes Dev 2001; 15: 1311–1333.

    Article  CAS  Google Scholar 

  43. Ma D, Nutt CL, Shanehsaz P, Peng X, Louis DN, Kaetzel DM . Autocrine PDGF-dependent gene expression in glioblastoma cells is mediated largely by activation of the transcription factor SRE-BP, and is associated with altered genotype and patient survival in human brain tumors. Cancer Res 2005; 65: 5523–5534.

    Article  CAS  Google Scholar 

  44. Liu Q, Muruve DA . Molecular basis of the inflammatory response to adenovirus vectors. Gene Ther 2003; 10: 935–940.

    Article  CAS  Google Scholar 

  45. Dong C, Davis RJ, Flavell RA . MAP kinases in the immune response. Annu Rev Immunol 2002; 20: 55–72 e-pub 4 October 2001: 55–72.

    Article  CAS  Google Scholar 

  46. Jackson DA, Juranek S, Lipps HJ . Designing nonviral vectors for efficient gene transfer and long-term gene expression. Mol Ther 2006; 14: 613–626.

    Article  CAS  Google Scholar 

  47. Lai CM, Lai YK, Rakoczy PE . Adenovirus and adeno-associated virus vectors. DNA Cell Biol 2002; 21: 895–913.

    Article  CAS  Google Scholar 

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Acknowledgements

We are grateful to Dr Ruth Keri and Dr John Nilson for providing DNA fragments harboring the glycoprotein α-subunit promoter. This study was supported by National Cancer Institute grant R01 CA83237, National Heart, Lung and Blood Institute grant R01 HL62877, and the Kentucky Lung Cancer Research Fund.

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Author notes

  1. A Mishra

    Present address: 4Current address: Department of Cancer Biology, School of Medicine, Johns Hopkins University, 1650 Orleans Street, Baltimore, MD 21231, USA.,

Authors and Affiliations

  1. Department of Molecular and Biomedical Pharmacology, College of Medicine, University of Kentucky, Lexington, KY, USA

    A Mishra, A K Ormerod, S D Kraner & D M Kaetzel

  2. Department of Pathology, College of Medicine, University of Kentucky, Lexington, KY, USA

    M L Cibull

  3. Department of Microbiology, Immunology and Molecular Genetics, College of Medicine, University of Kentucky, Lexington, KY, USA

    B T Spear

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  1. A Mishra
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Correspondence to D M Kaetzel.

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Supplementary Information accompanies the paper on Cancer Gene Therapy website (http://www.nature.com/cgt)

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Mishra, A., Ormerod, A., Cibull, M. et al. PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types. Cancer Gene Ther 16, 298–309 (2009). https://doi.org/10.1038/cgt.2008.92

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