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用于准确量化移植心脏中钠碘同向转运体（NIS）报告基因表达的非侵入性放射性碘成像。

Non-invasive radioiodine imaging for accurate quantitation of NIS reporter gene expression in transplanted hearts.

机构信息

Mayo Clinic William J von Liebig Transplant Center, Rochester, MN 55905, USA.

出版信息

Eur J Cardiothorac Surg. 2008 Jan;33(1):32-9. doi: 10.1016/j.ejcts.2007.09.033. Epub 2007 Nov 5.

DOI:10.1016/j.ejcts.2007.09.033
PMID:17980613
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2277515/
Abstract

OBJECTIVE

We studied the concordance of transgene expression in the transplanted heart using bicistronic adenoviral vector coding for a transgene of interest (human carcinoembryonic antigen: hCEA - beta human chorionic gonadotropin: betahCG) and for a marker imaging transgene (human sodium iodide symporter: hNIS).

METHODS

Inbred Lewis rats were used for syngeneic heterotopic cardiac transplantation. Donor rat hearts were perfused ex vivo for 30 min prior to transplantation with University of Wisconsin (UW) solution (n=3), with 10(9) pfu/ml of adenovirus expressing hNIS (Ad-NIS; n=6), hNIS-hCEA (Ad-NIS-CEA; n=6) and hNIS-betahCG (Ad-NIS-CG; n=6). On postoperative day (POD) 5, 10, 15 all animals underwent micro-single photon emission computed tomography/computed tomography (SPECT/CT) imaging of the donor hearts after tail vein injection of 1000 microCi (123)I and blood sample collection for hCEA and betahCG quantification.

RESULTS

Significantly higher image intensity was noted in the hearts perfused with Ad-NIS (1.1+/-0.2; 0.9+/-0.07), Ad-NIS-CEA (1.2+/-0.3; 0.9+/-0.1) and Ad-NIS-CG (1.1+/-0.1; 0.9+/-0.1) compared to UW group (0.44+/-0.03; 0.47+/-0.06) on POD 5 and 10 (p<0.05). Serum levels of hCEA and betahCG increased in animals showing high cardiac (123)I uptake, but not in those with lower uptake. Above this threshold, image intensities correlated well with serum levels of hCEA and betahCG (R(2)=0.99 and R(2)=0.96, respectively).

CONCLUSIONS

These data demonstrate that hNIS is an excellent reporter gene for the transplanted heart. The expression level of hNIS can be accurately and non-invasively monitored by serial radioisotopic SPECT imaging. High concordance has been demonstrated between imaging and soluble marker peptides at the maximum transgene expression on POD 5.

摘要

目的

我们使用编码感兴趣的转基因（人癌胚抗原：hCEA - β人绒毛膜促性腺激素：betahCG）和标记成像转基因（人钠碘同向转运体：hNIS）的双顺反子腺病毒载体，研究了移植心脏中转基因表达的一致性。

方法

近交系Lewis大鼠用于同基因异位心脏移植。在移植前，供体大鼠心脏用威斯康星大学（UW）溶液（n = 3）、10⁹ pfu/ml表达hNIS的腺病毒（Ad - NIS；n = 6）、hNIS - hCEA（Ad - NIS - CEA；n = 6）和hNIS - betahCG（Ad - NIS - CG；n = 6）进行离体灌注30分钟。在术后第5、10、15天，所有动物在尾静脉注射1000 μCi ¹²³I后对供体心脏进行微型单光子发射计算机断层扫描/计算机断层扫描（SPECT/CT）成像，并采集血样用于hCEA和betahCG定量。

结果

与UW组（0.44±0.03；0.47±0.06）相比，在术后第5天和第10天，用Ad - NIS（1.1±0.2；0.9±0.07）、Ad - NIS - CEA（1.2±0.3；0.9±0.1）和Ad - NIS - CG（1.1±0.1；0.9±0.1）灌注的心脏图像强度显著更高（p<0.05）。血清hCEA和betahCG水平在¹²³I摄取高的动物中升高，但在摄取低的动物中未升高。高于此阈值，图像强度与血清hCEA和betahCG水平相关性良好（R²分别为0.99和0.96）。

结论

这些数据表明hNIS是移植心脏的优良报告基因。hNIS的表达水平可通过连续放射性同位素SPECT成像进行准确、无创监测。在术后第5天最大转基因表达时，成像与可溶性标记肽之间已证明具有高度一致性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/f4a2d0113d60/nihms39320f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/c2ca8736bf95/nihms39320f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/63dfadca034b/nihms39320f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/bcbc7562a86c/nihms39320f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/f4a2d0113d60/nihms39320f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/c2ca8736bf95/nihms39320f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/63dfadca034b/nihms39320f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/bcbc7562a86c/nihms39320f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18e6/2277515/f4a2d0113d60/nihms39320f4.jpg

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