TY - JOUR
T1 - Endograft technology
T2 - A delivery vehicle for intravascular gene therapy
AU - Eton, Darwin
AU - Yu, Hong
AU - Wang, Yingcai
AU - Raines, Jeffrey
AU - Striker, Gary
AU - Livingstone, Alan
AU - Baxter, Timothy
N1 - Funding Information:
Supported by the American Heart Association (H.Y., D.E.); Pacific Vascular Foundation (D.E., H.Y.); Cordis Corp (D.E., H.Y.); David Kimmelman (D.E.); and The Norman F. Levy Foundation (D.E.).
PY - 2004/5
Y1 - 2004/5
N2 - Purpose: The purpose of this study was to determine whether vascular smooth muscle cells (SMCs) suffused into a bilayered stent graft retain and express a retrovirally transduced gene for 7 months in vivo. Method: SMCs harvested from dog jugular vein were retrovirally transduced to introduce genes for tissue plasminogen activator (t-PA) and β-galactosidase. These cells were then suffused into a novel dual-layered Dacron graft and cultured for 36 to 48 hours. The grafts were mounted on a Palmaz stent and balloon-expanded in the infrarenal aorta of the SMC donor dogs (n = 6). Grafts were recovered at 1, 2, 3, 4, 5, and 7 months. A control endograft suffused with SMCs transduced with only the β-galactosidase gene was placed in the dogs with grafts recovered at 2, 3, and 4 months. t-PA antigen concentration and expression were analyzed with an enzyme-linked immunosorbent assay. Results: Retained engineered SMCs (blue nuclei) were identified in the explanted grafts, neointima, and underlying aorta with X-gal staining. The t-PA antigen concentration and t-PA activity from the SMCs recovered from the grafts remained elevated for the duration of the experiment (7 months) at levels significantly higher (3.7 ± 0.2 ng/mL per 105 cells per 24 hours and 1.4 ± 0.1 IU/mL per 105 cells per 24 hours) than in control endografts (0.5 ± 0.03 ng/mL per 105 cells per 24 hours and 0.07 ± 0.00 IU/mL per 105 cells per 24 hours; P < .001). No graft stenosis was observed. Conclusion: Retrovirally engineered vascular SMCs survived the implantation trauma, repopulated each graft, migrated into the underlying aorta, and expressed the transduced genes for the 7-month duration of the experiment. This bilayered Dacron endograft model provides a platform to study direct intravascular gene therapy.
AB - Purpose: The purpose of this study was to determine whether vascular smooth muscle cells (SMCs) suffused into a bilayered stent graft retain and express a retrovirally transduced gene for 7 months in vivo. Method: SMCs harvested from dog jugular vein were retrovirally transduced to introduce genes for tissue plasminogen activator (t-PA) and β-galactosidase. These cells were then suffused into a novel dual-layered Dacron graft and cultured for 36 to 48 hours. The grafts were mounted on a Palmaz stent and balloon-expanded in the infrarenal aorta of the SMC donor dogs (n = 6). Grafts were recovered at 1, 2, 3, 4, 5, and 7 months. A control endograft suffused with SMCs transduced with only the β-galactosidase gene was placed in the dogs with grafts recovered at 2, 3, and 4 months. t-PA antigen concentration and expression were analyzed with an enzyme-linked immunosorbent assay. Results: Retained engineered SMCs (blue nuclei) were identified in the explanted grafts, neointima, and underlying aorta with X-gal staining. The t-PA antigen concentration and t-PA activity from the SMCs recovered from the grafts remained elevated for the duration of the experiment (7 months) at levels significantly higher (3.7 ± 0.2 ng/mL per 105 cells per 24 hours and 1.4 ± 0.1 IU/mL per 105 cells per 24 hours) than in control endografts (0.5 ± 0.03 ng/mL per 105 cells per 24 hours and 0.07 ± 0.00 IU/mL per 105 cells per 24 hours; P < .001). No graft stenosis was observed. Conclusion: Retrovirally engineered vascular SMCs survived the implantation trauma, repopulated each graft, migrated into the underlying aorta, and expressed the transduced genes for the 7-month duration of the experiment. This bilayered Dacron endograft model provides a platform to study direct intravascular gene therapy.
UR - http://www.scopus.com/inward/record.url?scp=2342599030&partnerID=8YFLogxK
U2 - 10.1016/j.jvs.2003.11.033
DO - 10.1016/j.jvs.2003.11.033
M3 - Article
C2 - 15111863
AN - SCOPUS:2342599030
SN - 0741-5214
VL - 39
SP - 1066
EP - 1073
JO - Journal of Vascular Surgery
JF - Journal of Vascular Surgery
IS - 5
ER -