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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1995 Jun 20;92(13):5855–5859. doi: 10.1073/pnas.92.13.5855

A gene therapy strategy using a transcription factor decoy of the E2F binding site inhibits smooth muscle proliferation in vivo.

R Morishita 1, G H Gibbons 1, M Horiuchi 1, K E Ellison 1, M Nakama 1, L Zhang 1, Y Kaneda 1, T Ogihara 1, V J Dzau 1
PMCID: PMC41600  PMID: 7597041

Abstract

The application of DNA technology to regulate the transcription of disease-related genes in vivo has important therapeutic potentials. The transcription factor E2F plays a pivotal role in the coordinated transactivation of cell cycle-regulatory genes such as c-myc, cdc2, and the gene encoding proliferating-cell nuclear antigen (PCNA) that are involved in lesion formation after vascular injury. We hypothesized that double-stranded DNA with high affinity for E2F may be introduced in vivo as a decoy to bind E2F and block the activation of genes mediating cell cycle progression and intimal hyperplasia after vascular injury. Gel mobility-shift assays showed complete competition for E2F binding protein by the E2F decoy. Transfection with E2F decoy inhibited expression of c-myc, cdc2, and the PCNA gene as well as vascular smooth muscle cell proliferation both in vitro and in the in vivo model of rat carotid injury. Furthermore, 2 weeks after in vivo transfection, neointimal formation was significantly prevented by the E2F decoy, and this inhibition continued up to 8 weeks after a single transfection in a dose-dependent manner. Transfer of an E2F decoy can therefore modulate gene expression and inhibit smooth muscle proliferation and vascular lesion formation in vivo.

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Selected References

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  1. Bielinska A., Shivdasani R. A., Zhang L. Q., Nabel G. J. Regulation of gene expression with double-stranded phosphorothioate oligonucleotides. Science. 1990 Nov 16;250(4983):997–1000. doi: 10.1126/science.2237444. [DOI] [PubMed] [Google Scholar]
  2. Casscells W. Migration of smooth muscle and endothelial cells. Critical events in restenosis. Circulation. 1992 Sep;86(3):723–729. doi: 10.1161/01.cir.86.3.723. [DOI] [PubMed] [Google Scholar]
  3. Chittenden T., Livingston D. M., Kaelin W. G., Jr The T/E1A-binding domain of the retinoblastoma product can interact selectively with a sequence-specific DNA-binding protein. Cell. 1991 Jun 14;65(6):1073–1082. doi: 10.1016/0092-8674(91)90559-h. [DOI] [PubMed] [Google Scholar]
  4. Dalton S. Cell cycle regulation of the human cdc2 gene. EMBO J. 1992 May;11(5):1797–1804. doi: 10.1002/j.1460-2075.1992.tb05231.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gibbons G. H., Dzau V. J. The emerging concept of vascular remodeling. N Engl J Med. 1994 May 19;330(20):1431–1438. doi: 10.1056/NEJM199405193302008. [DOI] [PubMed] [Google Scholar]
  6. Hayashi K., Makino R., Kawamura H., Arisawa A., Yoneda K. Characterization of rat c-myc and adjacent regions. Nucleic Acids Res. 1987 Aug 25;15(16):6419–6436. doi: 10.1093/nar/15.16.6419. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Helin K., Lees J. A., Vidal M., Dyson N., Harlow E., Fattaey A. A cDNA encoding a pRB-binding protein with properties of the transcription factor E2F. Cell. 1992 Jul 24;70(2):337–350. doi: 10.1016/0092-8674(92)90107-n. [DOI] [PubMed] [Google Scholar]
  8. Hermans W. R., Rensing B. J., Strauss B. H., Serruys P. W. Prevention of restenosis after percutaneous transluminal coronary angioplasty: the search for a "magic bullet". Am Heart J. 1991 Jul;122(1 Pt 1):171–187. doi: 10.1016/0002-8703(91)90775-d. [DOI] [PubMed] [Google Scholar]
  9. Hiebert S. W., Lipp M., Nevins J. R. E1A-dependent trans-activation of the human MYC promoter is mediated by the E2F factor. Proc Natl Acad Sci U S A. 1989 May;86(10):3594–3598. doi: 10.1073/pnas.86.10.3594. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Horiuchi M., Nakamura N., Tang S. S., Barrett G., Dzau V. J. Molecular mechanism of tissue-specific regulation of mouse renin gene expression by cAMP. Identification of an inhibitory protein that binds nuclear transcriptional factor. J Biol Chem. 1991 Aug 25;266(24):16247–16254. [PubMed] [Google Scholar]
  11. Kaelin W. G., Jr, Krek W., Sellers W. R., DeCaprio J. A., Ajchenbaum F., Fuchs C. S., Chittenden T., Li Y., Farnham P. J., Blanar M. A. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell. 1992 Jul 24;70(2):351–364. doi: 10.1016/0092-8674(92)90108-o. [DOI] [PubMed] [Google Scholar]
  12. Kim Y. K., Lee A. S. Identification of a 70-base-pair cell cycle regulatory unit within the promoter of the human thymidine kinase gene and its interaction with cellular factors. Mol Cell Biol. 1991 Apr;11(4):2296–2302. doi: 10.1128/mcb.11.4.2296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Klein-Hitpass L., Tsai S. Y., Weigel N. L., Allan G. F., Riley D., Rodriguez R., Schrader W. T., Tsai M. J., O'Malley B. W. The progesterone receptor stimulates cell-free transcription by enhancing the formation of a stable preinitiation complex. Cell. 1990 Jan 26;60(2):247–257. doi: 10.1016/0092-8674(90)90740-6. [DOI] [PubMed] [Google Scholar]
  14. Kovesdi I., Reichel R., Nevins J. R. Identification of a cellular transcription factor involved in E1A trans-activation. Cell. 1986 Apr 25;45(2):219–228. doi: 10.1016/0092-8674(86)90386-7. [DOI] [PubMed] [Google Scholar]
  15. Montalescot G., Faraggi M., Drobinski G., Messian O., Evans J., Grosgogeat Y., Thomas D. Myocardial viability in patients with Q wave myocardial infarction and no residual ischemia. Circulation. 1992 Jul;86(1):47–55. doi: 10.1161/01.cir.86.1.47. [DOI] [PubMed] [Google Scholar]
  16. Morishita R., Gibbons G. H., Ellison K. E., Nakajima M., Zhang L., Kaneda Y., Ogihara T., Dzau V. J. Single intraluminal delivery of antisense cdc2 kinase and proliferating-cell nuclear antigen oligonucleotides results in chronic inhibition of neointimal hyperplasia. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8474–8478. doi: 10.1073/pnas.90.18.8474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morishita R., Gibbons G. H., Kaneda Y., Ogihara T., Dzau V. J. Novel and effective gene transfer technique for study of vascular renin angiotensin system. J Clin Invest. 1993 Jun;91(6):2580–2585. doi: 10.1172/JCI116496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nevins J. R. E2F: a link between the Rb tumor suppressor protein and viral oncoproteins. Science. 1992 Oct 16;258(5081):424–429. doi: 10.1126/science.1411535. [DOI] [PubMed] [Google Scholar]
  19. Popma J. J., Califf R. M., Topol E. J. Clinical trials of restenosis after coronary angioplasty. Circulation. 1991 Sep;84(3):1426–1436. doi: 10.1161/01.cir.84.3.1426. [DOI] [PubMed] [Google Scholar]
  20. Sullenger B. A., Gallardo H. F., Ungers G. E., Gilboa E. Overexpression of TAR sequences renders cells resistant to human immunodeficiency virus replication. Cell. 1990 Nov 2;63(3):601–608. doi: 10.1016/0092-8674(90)90455-n. [DOI] [PubMed] [Google Scholar]
  21. Thalmeier K., Synovzik H., Mertz R., Winnacker E. L., Lipp M. Nuclear factor E2F mediates basic transcription and trans-activation by E1a of the human MYC promoter. Genes Dev. 1989 Apr;3(4):527–536. doi: 10.1101/gad.3.4.527. [DOI] [PubMed] [Google Scholar]
  22. Wagner S., Green M. R. Retinoblastoma. A transcriptional tryst. Nature. 1991 Jul 18;352(6332):189–190. doi: 10.1038/352189a0. [DOI] [PubMed] [Google Scholar]
  23. Watson R. J., Dyson P. J., McMahon J. Multiple c-myb transcript cap sites are variously utilized in cells of mouse haemopoietic origin. EMBO J. 1987 Jun;6(6):1643–1651. doi: 10.1002/j.1460-2075.1987.tb02413.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Weintraub S. J., Prater C. A., Dean D. C. Retinoblastoma protein switches the E2F site from positive to negative element. Nature. 1992 Jul 16;358(6383):259–261. doi: 10.1038/358259a0. [DOI] [PubMed] [Google Scholar]
  25. Yamaguchi M., Hayashi Y., Hirose F., Matsuoka S., Shiroki K., Matsukage A. Activation of the mouse proliferating cell nuclear antigen gene promoter by adenovirus type 12 E1A proteins. Jpn J Cancer Res. 1992 Jun;83(6):609–617. doi: 10.1111/j.1349-7006.1992.tb00133.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yee A. S., Reichel R., Kovesdi I., Nevins J. R. Promoter interaction of the E1A-inducible factor E2F and its potential role in the formation of a multi-component complex. EMBO J. 1987 Jul;6(7):2061–2068. doi: 10.1002/j.1460-2075.1987.tb02471.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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