Venous bypass grafts are useful treatments for obstructive coronary artery disease. However, their usefulness is limited by accelerated atherosclerosis. Genetic engineering of venous bypass grafts that prevented atherosclerosis could improve long-term graft patency and clinical outcomes. We used a rabbit model of jugular vein-to-carotid interposition grafting to develop gene therapy for vein-graft atherosclerosis. Rabbit veins were easily transduced in situ with a first-generation adenoviral vector; however, most transgene expression (∼80%) was lost within 3 days after arterial grafting. This rapid loss of transgene expression was not prevented by transducing veins after grafting or by prolonged ex vivo transduction. However, delaying vein-graft transduction for 28 days (after the vein had adapted to the arterial circulation) prevented this early loss of transgene expression. We used the delayed transduction approach to test the durability of expression of a therapeutic transgene (apolipoprotein A-I) expressed from a helper-dependent adenoviral (HDAd) vector. HDAd DNA and apolipoprotein A-I mRNA were easily detectable in transduced vein grafts. Vector DNA and mRNA declined by 4 weeks, and then persisted stably for at least 6 months. Delaying transduction for 28 days after grafting permitted initiation of vein-graft neointimal growth and medial thickening before gene transfer. However, vein-graft lumen diameter was not compromised, because of gradual outward remodeling of grafted veins. Our data highlight the promise of HDAd-mediated gene therapy, delivered to arterialized vein grafts, for preventing vein-graft atherosclerosis.
Get full access to this article
View all access options for this article.
References
1.
GoAS, MozaffarianD, RogerVL, et al.Heart disease and stroke statistics—2013 update: a report from the American Heart Association. Circulation, 2013; 127:e6–e245.
2.
HlatkyMA, BoothroydDB, BravataDM, et al.Coronary artery bypass surgery compared with percutaneous coronary interventions for multivessel disease: a collaborative analysis of individual patient data from ten randomised trials. Lancet, 2009; 373:1190–1197.
3.
SerruysPW, MoriceMC, KappeteinAP, et al.Percutaneous coronary intervention versus coronary-artery bypass grafting for severe coronary artery disease. N Engl J Med, 2009; 360:961–972.
4.
MohrFW, MoriceMC, KappeteinAP, et al.Coronary artery bypass graft surgery versus percutaneous coronary intervention in patients with three-vessel disease and left main coronary disease: 5-year follow-up of the randomised, clinical SYNTAX trial. Lancet, 2013; 381:629–638.
5.
HarskampRE, LopesRD, BaisdenCE, et al.Saphenous vein graft failure after coronary artery bypass surgery: pathophysiology, management, and future directions. Ann Surg, 2013; 257:824–833.
6.
TatoulisJ, BuxtonBF, and FullerJA. Patencies of 2127 arterial to coronary conduits over 15 years. Ann Thorac Surg, 2004; 77:93–101.
7.
MehtaRH, FergusonTB, LopesRD, et al.Saphenous vein grafts with multiple versus single distal targets in patients undergoing coronary artery bypass surgery: one-year graft failure and five-year outcomes from the Project of Ex-Vivo Vein Graft Engineering via Transfection (PREVENT) IV trial. Circulation, 2011; 124:280–288.
GoldmanS, CopelandJ, MoritzT, et al.Long-term graft patency (3 years) after coronary artery surgery. Effects of aspirin: results of a VA Cooperative study. Circulation, 1994; 89:1138–1143.
10.
Post Coronary Bypass Graft Trial Investigators. The effect of aggressive lowering of low-density lipoprotein cholesterol levels and low-dose anticoagulation on obstructive changes in saphenous-vein coronary-artery bypass grafts. N Engl J Med, 1997; 336:153–162.
11.
SoutherlandKW, FrazierSB, BowlesDE, et al.Gene therapy for the prevention of vein graft disease. Transl Res, 2013; 161:321–338.
12.
LopesRD, MehtaRH, HafleyGE, et al.Relationship between vein graft failure and subsequent clinical outcomes after coronary artery bypass surgery. Circulation, 2012; 125:749–756.
HarskampRE, BeijkMA, DammanP, et al.Clinical outcome after surgical or percutaneous revascularization in coronary bypass graft failure. J Cardiovasc Med (Hagerstown), 2013; 14:438–445.
15.
BradshawAC, and BakerAH. Gene therapy for cardiovascular disease: perspectives and potential. Vascul Pharmacol, 2013; 58:174–181.
16.
MotwaniJG, and TopolEJ. Aortocoronary saphenous vein graft disease: pathogenesis, predisposition, and prevention. Circulation, 1998; 97:916–931.
17.
ContiVR, and HunterGC. Gene therapy and vein graft patency in coronary artery bypass graft surgery. JAMA, 2005; 294:2495–2497.
18.
FlynnR, QianK, TangC, et al.Expression of apolipoprotein A-I in rabbit carotid endothelium protects against atherosclerosis. Mol Ther, 2011; 19:1833–1841.
19.
VassalliG, AgahR, QiaoR, et al.A mouse model of arterial gene transfer: antigen-specific immunity is a minor determinant of the early loss of adenovirus-mediated transgene expression. Circ Res, 1999; 85:e25–e32.
20.
ParksRJ, ChenL, AntonM, et al.A helper-dependent adenovirus vector system: removal of helper virus by Cre-mediated excision of the viral packaging signal. Proc Natl Acad Sci U S A, 1996; 93:13565–13570.
21.
FlynnR, BucklerJM, TangC, et al.Helper-dependent adenoviral vectors are superior in vitro to first-generation vectors for endothelial cell-targeted gene therapy. Mol Ther, 2010; 18:2121–2129.
22.
ChenL, AntonM, and GrahamFL. Production and characterization of human 293 cell lines expressing the site-specific recombinase Cre. Somat Cell Mol Genet, 1996; 22:477–488.
23.
DronadulaN, DuL, FlynnR, et al.Construction of a novel expression cassette for increasing transgene expression in vivo in endothelial cells of large blood vessels. Gene Ther, 2011; 18:501–508.
24.
MitterederN, MarchKL, and TrapnellBC. Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol, 1996; 70:7498–7509.
25.
DuL, DronadulaN, TanakaS, et al.Helper-dependent adenoviral vector achieves prolonged, stable expression of interleukin-10 in rabbit carotid arteries but does not limit early atherogenesis. Hum Gene Ther, 2011; 22:959–968.
26.
JiangZ, WuL, MillerBL, et al.A novel vein graft model: adaptation to differential flow environments. Am J Physiol Heart Circ Physiol, 2004; 286:H240–H245.
27.
ZwolakRM, KirkmanTR, and ClowesAW. Atherosclerosis in rabbit vein grafts. Arteriosclerosis, 1989; 9:374–379.
28.
SchneiderDB, VassalliG, WenS, et al.Expression of Fas ligand in arteries of hypercholesterolemic rabbits accelerates atherosclerotic lesion formation. Arterioscler Thromb Vasc Biol, 2000; 20:298–308.
29.
ZwolakRM, AdamsMC, and ClowesAW. Kinetics of vein graft hyperplasia: association with tangential stress. J Vasc Surg, 1987; 5:126–136.
30.
RobertsonKE, McDonaldRA, OldroydKG, et al.Prevention of coronary in-stent restenosis and vein graft failure: does vascular gene therapy have a role?. Pharmacol Ther, 2012; 136:23–34.
31.
ChannonKM, FultonGJ, GrayJL, et al.Efficient adenoviral gene transfer to early venous bypass grafts: comparison with native vessels. Cardiovasc Res, 1997; 35:505–513.
32.
LemarchandP, JonesM, YamadaI, et al.In vivo gene transfer and expression in normal uninjured blood vessels using replication-deficient recombinant adenovirus vectors. Circ Res, 1993; 72:1132–1138.
33.
ChannonKM, QianH, YoungbloodSA, et al.Acute host-mediated endothelial injury after adenoviral gene transfer in normal rabbit arteries: impact on transgene expression and endothelial function. Circ Res, 1998; 82:1253–1262.
34.
WanS, GeorgeSJ, NicklinSA, et al.Overexpression of p53 increases lumen size and blocks neointima formation in porcine interposition vein grafts. Mol Ther, 2004; 9:689–698.
35.
DaviesMG, HuynhTT, FultonGJ, et al.G protein signaling and vein graft intimal hyperplasia: reduction of intimal hyperplasia in vein grafts by a Gβγ inhibitor suggests a major role of G protein signaling in lesion development. Arterioscler Thromb Vasc Biol, 1998; 18:1275–1280.
36.
KibbeMR, TzengE, GleixnerSL, et al.Adenovirus-mediated gene transfer of human inducible nitric oxide synthase in porcine vein grafts inhibits intimal hyperplasia. J Vasc Surg, 2001; 34:156–165.
37.
WestNE, QianH, GuzikTJ, et al.Nitric oxide synthase (nNOS) gene transfer modifies venous bypass graft remodeling: effects on vascular smooth muscle cell differentiation and superoxide production. Circulation, 2001; 104:1526–1532.
SarjeantJM, and RabinovitchM. Understanding and treating vein graft atherosclerosis. Cardiovasc Pathol, 2002; 11:263–271.
40.
AlexanderJH, HafleyG, HarringtonRA, et al.Efficacy and safety of edifoligide, an E2F transcription factor decoy, for prevention of vein graft failure following coronary artery bypass graft surgery: PREVENT IV: a randomized controlled trial. JAMA, 2005; 294:2446–2454.
41.
ConteMS, BandykDF, ClowesAW, et al.Results of PREVENT III: a multicenter, randomized trial of edifoligide for the prevention of vein graft failure in lower extremity bypass surgery. J Vasc Surg, 2006; 43:742–751; discussion 751.
42.
KilianEG, EifertS, Beiras-FernandezA, et al.Adeno-associated virus-mediated gene transfer in a rabbit vein graft model. Circ J, 2008; 72:1700–1704.
43.
EeftingD, BotI, De VriesMR, et al.Local lentiviral short hairpin RNA silencing of CCR2 inhibits vein graft thickening in hypercholesterolemic apolipoprotein E3-Leiden mice. J Vasc Surg, 2009; 50:152–160.
44.
WenS, GrafS, MasseyPG, et al.Improved vascular gene transfer with a helper-dependent adenoviral vector. Circulation, 2004; 110:1484–1491.
45.
JiangB, QianK, DuL, et al.Helper-dependent adenovirus is superior to first-generation adenovirus for expressing transgenes in atherosclerosis-prone arteries. Arterioscler Thromb Vasc Biol, 2011; 31:1317–1325.
46.
SchulickAH, VassalliG, DunnPF, et al.Established immunity precludes adenovirus-mediated gene transfer in rat carotid arteries: potential for immunosuppression and vector engineering to overcome barriers of immunity. J Clin Invest, 1997; 99:209–219.
47.
FlugelmanMY, JaklitschMT, NewmanKD, et al.Low level in vivo gene transfer into the arterial wall through a perforated balloon catheter. Circulation, 1992; 85:1110–1117.
48.
RaperSE, ChirmuleN, LeeFS, et al.Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab, 2003; 80:148–158.
MannMJ, GibbonsGH, KernoffRS, et al.Genetic engineering of vein grafts resistant to atherosclerosis. Proc Natl Acad Sci U S A, 1995; 92:4502–4506.
51.
SchwartzSM, DebloisD, and O'BrienERM. The intima: soil for atherosclerosis and restenosis. Circ Res, 1995; 77:445–465.
52.
NewbyAC, and BakerAH. Targets for gene therapy of vein grafts. Curr Opin Cardiol, 1999; 14:489–494.
53.
RaderDJ, AlexanderET, WeibelGL, et al.The role of reverse cholesterol transport in animals and humans and relationship to atherosclerosis. J Lipid Res, 2009; 50(Suppl.):S189–S194.
54.
ChenS-J, WilsonJM, and MullerDWM. Adenovirus-mediated gene transfer of soluble vascular cell adhesion molecule to porcine interposition vein grafts. Circulation, 1994; 89:1922–1928.
55.
TurunenP, PuhakkaHL, HeikuraT, et al.Extracellular superoxide dismutase with vaccinia virus anti-inflammatory protein 35K or tissue inhibitor of metalloproteinase-1: combination gene therapy in the treatment of vein graft stenosis in rabbits. Hum Gene Ther, 2006; 17:405–414.
56.
NewmanKD, DunnPF, OwensJW, et al.Adenovirus-mediated gene transfer into normal rabbit arteries results in prolonged vascular cell activation, inflammation, and neointimal hyperplasia. J Clin Invest, 1995; 96:2955–2965.
57.
LafontA, LoirandG, PacaudP, et al.Vasomotor dysfunction early after exposure of normal rabbit arteries to an adenoviral vector. Hum Gene Ther, 1997; 8:1033–1040.
58.
LonnU, PeterzenB, GranfeldtH, et al.Coronary artery operation supported by the Hemopump: an experimental study on pig. Ann Thorac Surg, 1994; 58:516–518.
59.
ArmstrongML, and HeistadDD. Animal models of atherosclerosis. Atherosclerosis, 1990; 85:15–23.
60.
AngeliniGD, IzzatMB, BryanAJ, et al.External stenting reduces early medial and neointimal thickening in a pig model of arteriovenous bypass grafting. J Thorac Cardiovasc Surg, 1996; 112:79–84.
61.
SchachnerT, LauferG, and BonattiJ. In vivo (animal) models of vein graft disease. Eur J Cardiothorac Surg, 2006; 30:451–463.
62.
CremersB, MilewskiK, CleverYP, et al.Long-term effects on vascular healing of bare metal stents delivered via paclitaxel-coated balloons in the porcine model of restenosis. Catheter Cardiovasc Interv, 2012; 80:603–610.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
