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In Vivo Human MCP-1 Transfection in Porcine Arteries by Intravascular Electroporation

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Purpose

The purpose of this study was to develop a nonviral gene transfer method for therapeutic delivery of the human monocyte chemoattractant protein-1 (MCP-1) in patients with peripheral artery disease, using local catheter-mediated electrotransfer of naked plasmid DNA into arteries.

Methods

Arterial walls of the A. profunda femoris of pigs were transfected either with a human MCP-1 or with a firefly luciferase-encoding DNA construct. The efficacy of electrotransfer of DNA was analyzed after 2 days by quantitative polymerase chain reaction (PCR) or luciferase activity measurements. To optimize MCP-1 gene transfer conditions, a voltage range of 60–150 V was applied as a train of six square pulses of 20 ms each at 1 Hz and was combined with a dose of 150 μg DNA. Subsequently, the optimized voltage was used to test a dose range of 80–300 μg DNA.

Results

The voltage optimum for arterial transfection was observed at 80 volts. Using this setting, the dose application of 300 μg MCP-1 plasmid DNA (the maximal dose tested) demonstrated the highest MCP-1 expression signal. The electric pulses and the transfer and expression of human MCP-1 per se did not induce endogenous porcine MCP-1 expression in treated arteries. Interestingly, angioplastic predilation of the artery before gene transfer, which had originally been postulated to enhance transfection by improving access of the plasmid to subendothelial cell layers, resulted in an attenuated transfection efficacy.

Conclusions

The present study demonstrates that transluminal catheter-based electroporation provides an efficient technology for nonviral intravascular gene transfer by just applying unformulated DNA.

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Abbreviations

EP catheter:

electroporation catheter

FAM:

6-carboxy-fluorescein

GAPDH:

glycerolaldehyde-3-phosphate dehydrogenase

JOE:

6-carboxy-4′,5′-dichloro-2′,7′-dimethoxy-fluorescein

ms:

milliseconds

RLU:

relative light units

TAMRA:

6-carboxytetramethylrhodamine

References

  1. C. Diehm A. Schuster J. R. Allenberg H. Darius R. Haberl S. Lange D. Pittrow B. Stritzky Particlevon G. Tepohl H. J. Trampisch (2004) ArticleTitleHigh prevalence of peripheral arterial disease and co-morbidity in 6880 primary care patients: cross-sectional study Atherosclerosis 172 95–105 Occurrence Handle10.1016/S0021-9150(03)00204-1 Occurrence Handle14709362

    Article  PubMed  Google Scholar 

  2. A. T. Hirsch M. H. Criqui D. Treat-Jacobson J. G. Regensteiner M. A. Creager J. W. Olin S. H. Krook D. B. Hunninghake A. J. Comerota M. E. Walsh M. M. McDermott W. R. Hiatt (2001) ArticleTitlePeripheral arterial disease detection, awareness, and treatment in primary care JAMA 286 1317–1324 Occurrence Handle10.1001/jama.286.11.1317 Occurrence Handle11560536

    Article  PubMed  Google Scholar 

  3. W. T. Meijer A. W. Hoes D. Rutgers M. L. Bots A. Hofman D. E. Grobbee (1998) ArticleTitlePeripheral arterial disease in the elderly: The Rotterdam Study Arterioscler. Thromb. Vasc. Biol. 18 185–192 Occurrence Handle9484982

    PubMed  Google Scholar 

  4. H. E. Stoffers P. E. Rinkens A. D. Kester V. Kaiser J. A. Knottnerus (1996) ArticleTitleThe prevalence of asymptomatic and unrecognized peripheral arterial occlusive disease Int. J. Epidemiol. 25 282–290 Occurrence Handle9119553

    PubMed  Google Scholar 

  5. W. R. Hiatt (2002) ArticleTitlePharmacologic therapy for peripheral arterial disease and claudication J. Vasc. Surg. 36 1283–1291 Occurrence Handle10.1067/mva.2002.129654 Occurrence Handle12469066

    Article  PubMed  Google Scholar 

  6. J. G. Regensteiner W. R. Hiatt (2002) ArticleTitleCurrent medical therapies for patients with peripheral arterial disease: a critical review Am. J. Med. 112 49–57 Occurrence Handle10.1016/S0002-9343(01)01034-8 Occurrence Handle11812407

    Article  PubMed  Google Scholar 

  7. Y. J. Shyy H. J. Hsieh S. Usami S. Chien (1994) ArticleTitleFluid shear stress induces a biphasic response of human monocyte chemotactic protein 1 gene expression in vascular endothelium Proc. Natl. Acad. Sci. USA 91 4678–4682 Occurrence Handle8197118

    PubMed  Google Scholar 

  8. M. Arras W. D. Ito D. Scholz B. Winkler J. Schaper W. Schaper (1998) ArticleTitleMonocyte activation in angiogenesis and collateral growth in the rabbit hindlimb J. Clin. Invest. 101 40–50 Occurrence Handle9421464

    PubMed  Google Scholar 

  9. P. J. Polverini P. S. Cotran M. A. Gimbrone SuffixJr. E. R. Unanue (1977) ArticleTitleActivated macrophages induce vascular proliferation Nature 269 804–806 Occurrence Handle10.1038/269804a0 Occurrence Handle927505

    Article  PubMed  Google Scholar 

  10. W. D. Ito M. Arras B. Winkler D. Scholz J. Schaper W. Schaper (1997) ArticleTitleMonocyte chemotactic protein-1 increases collateral and peripheral conductance after femoral artery occlusion Circ. Res. 80 829–837 Occurrence Handle9168785

    PubMed  Google Scholar 

  11. E. Deindl I. Buschmann I. E. Hoefer T. Podzuweit K. Boengler S. Vogel N. Royen Particlevan B. Fernandez W. Schaper (2001) ArticleTitleRole of ischemia and of hypoxia-inducible genes in arteriogenesis after femoral artery occlusion in the rabbit Circ. Res. 89 779–786 Occurrence Handle11679407

    PubMed  Google Scholar 

  12. H. Kamihata H. Matsubara T. Nishiue S. Fujiyama Y. Tsutsumi R. Ozono H. Masaki Y. Mori O. Iba E. Tateishi A. Kosaki S. Shintani T. Murohara T. Imaizumi T. Iwasaka (2001) ArticleTitleImplantation of bone marrow mononuclear cells into ischemic myocardium enhances collateral perfusion and regional function via side supply of angioblasts, angiogenic ligands, and cytokines Circulation 104 1046–1052 Occurrence Handle11524400

    PubMed  Google Scholar 

  13. F. C. White S. M. Carroll A. Magnet C. M. Bloor (1992) ArticleTitleCoronary collateral development in swine after coronary artery occlusion Circ. Res. 71 1490–1500 Occurrence Handle1423941

    PubMed  Google Scholar 

  14. I. E. Hoefer N. Royen Particlevan I. R. Buschmann J. J. Piek W. Schaper (2001) ArticleTitleTime course of arteriogenesis following femoral artery occlusion in the rabbit Cardiovasc. Res. 49 609–617 Occurrence Handle10.1016/S0008-6363(00)00243-1 Occurrence Handle11166274

    Article  PubMed  Google Scholar 

  15. M. Voskuil N. Royen Particlevan I. E. Hoefer R. Seidler B. D. Guth C. Bode W. Schaper J. J. Piek I. R. Buschmann (2003) ArticleTitleModulation of collateral artery growth in a porcine hindlimb ligation model using MCP-1 Am. J. Physiol., Heart Circ. Physiol. 284 H1422–H1428

    Google Scholar 

  16. M. A. Azrin J. F. Mitchel D. B. Fram C. A. Pedersen R. W. Cartun J. J. Barry L. M. Bow D. D. Waters R. G. McKay (1994) ArticleTitleDecreased platelet deposition and smooth muscle cell proliferation after intramural heparin delivery with hydrogel-coated balloons Circulation 90 433–441 Occurrence Handle8026030

    PubMed  Google Scholar 

  17. H. Wolinsky C. S. Lin (1991) ArticleTitleUse of the perforated balloon catheter to infuse marker substances into diseased coronary artery walls after experimental postmortem angioplasty J. Am. Coll. Cardiol. 17 174B–178B Occurrence Handle1707901

    PubMed  Google Scholar 

  18. P. M. Consigny J. J. Barry N. J. Vitali (1994) ArticleTitleLocal delivery of an antiproliferative drug with use of hydrogel-coated angioplasty balloons J. Vasc. Interv. Radiol. 5 553–560 Occurrence Handle7949710

    PubMed  Google Scholar 

  19. R. Riessen J. M. Isner (1994) ArticleTitleProspects for site-specific delivery of pharmacologic and molecular therapies J. Am. Coll. Cardiol. 23 1234–1244 Occurrence Handle8144794

    PubMed  Google Scholar 

  20. S. M. Schwartz. Restenosis Summit VII. Cleveland, OH, pp. 290–294

  21. D. Rabussay N. B. Dev J. Fewell L. C. Smith G. Widera L. Zhang (2003) ArticleTitleEnhancement of therapeutic drug and DNA delivery into cells by electroporation J. Phys., D, Appl. Phys. 36 348–363

    Google Scholar 

  22. N. B. Dev M. Joseph D. Rabussay (2002) ArticleTitleEfficient in vivo transgene expression in the vascular wall driven by endovascular electroporation Mol. Ther. 5 S240

    Google Scholar 

  23. J. C. Wu M. Inubushi G. Sundaresan H. R. Schelbert S. S. Gambhir (2002) ArticleTitleOptical imaging of cardiac reporter gene expression in living rats Circulation 105 1631–1634 Occurrence Handle10.1161/01.CIR.0000014984.95520.AD Occurrence Handle11940538

    Article  PubMed  Google Scholar 

  24. A. Muhs M. C. Lenter R. W. Seidler R. Zweigerdt M. Kirchengast R. Weser M. Ruediger B. Guth (2004) ArticleTitleNonviral monocyte chemoattractant protein-1 gene transfer improves arteriogenesis after femoral artery occlusion Gene Ther. 11 1685–1693 Occurrence Handle10.1038/sj.gt.3302360 Occurrence Handle15470479

    Article  PubMed  Google Scholar 

  25. C. E. Thomas A. Ehrhardt M. A. Kay (2003) ArticleTitleProgress and problems with the use of viral vectors for gene therapy Nat. Rev. Genet. 4 346–358 Occurrence Handle10.1038/nrg1066 Occurrence Handle12728277

    Article  PubMed  Google Scholar 

  26. Q. Liu D. A. Muruve (2003) ArticleTitleMolecular basis of the inflammatory response to adenovirus vectors Gene Ther. 10 935–940 Occurrence Handle10.1038/sj.gt.3302036 Occurrence Handle12756413

    Article  PubMed  Google Scholar 

  27. S. A. Audouy L. F. Leij Particlede D. Hoekstra G. Molema (2002) ArticleTitleIn vivo characteristics of cationic liposomes as delivery vectors for gene therapy Pharm. Res. 19 1599–1605 Occurrence Handle10.1023/A:1020989709019 Occurrence Handle12458664

    Article  PubMed  Google Scholar 

  28. J. Cui K. A. Robinson J. E. Brown N. A. F. Chronos G. D. Cipolla N. B. Dev G. Hofmann N. Crowford S. B. King (1997) ArticleTitleLocal drug delivery to pig carotid arteries by direct vessel wall electroporation using novel catheter J. Am. Coll. Cardiol. 29 749

    Google Scholar 

  29. T. Matsumoto K. Komori T. Shoji S. Kuma M. Kume T. Yamaoka E. Mori T. Furuyama Y. Yonemitsu K. Sugimachi (2001) ArticleTitleSuccessful and optimized in vivo gene transfer to rabbit carotid artery mediated by electronic pulse Gene Ther. 8 1174–1179 Occurrence Handle10.1038/sj.gt.3301502 Occurrence Handle11509948

    Article  PubMed  Google Scholar 

  30. F. Wempe V. Lindner H. G. Augustin (1997) ArticleTitleBasic fibroblast growth factor (bFGF) regulates the expression of the CC chemokine monocyte chemoattractant protein-1 (MCP-1) in autocrine-activated endothelial cells Arterioscler. Thromb. Vasc. Biol. 17 2471–2478 Occurrence Handle9409217

    PubMed  Google Scholar 

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Acknowledgments

The expert technical assistance of Dunja Nitz is gratefully acknowledged. We thank Dr. Markus Wolfram for his support preparing the manuscript. Dr. Barbara Enenkel is acknowledged for design and preparation of the reporter gene vector.

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Authors and Affiliations

  1. Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, Connecticut, 06877, USA

    Randolph W. Seidler

  2. Boehringer Ingelheim Pharma GmbH & Co. KG, Birkendorfer Str. 65, 88397, Biberach, Germany

    Susanne Allgäuer, Susanne Ailinger, Andreas Sterner, Henri Doods & Martin C. Lenter

  3. Genetronics Inc., San Diego, California, 92121, USA

    Nagendu Dev & Dietmar Rabussay

Authors
  1. Randolph W. Seidler
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  2. Susanne Allgäuer
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  3. Susanne Ailinger
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  4. Andreas Sterner
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  5. Nagendu Dev
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  6. Dietmar Rabussay
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  7. Henri Doods
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  8. Martin C. Lenter
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Corresponding author

Correspondence to Martin C. Lenter.

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Seidler, R.W., Allgäuer, S., Ailinger, S. et al. In Vivo Human MCP-1 Transfection in Porcine Arteries by Intravascular Electroporation. Pharm Res 22, 1685–1691 (2005). https://doi.org/10.1007/s11095-005-6334-9

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  • DOI: https://doi.org/10.1007/s11095-005-6334-9

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