Zohlnhöfer D, Klein C A, Richter T, Brandl R, Murr A, Nührenberg T, Schömig A, Baeuerle P A, Neumann F J
Micromet AG, München, Germany.
Circulation. 2001 Mar 13;103(10):1396-402. doi: 10.1161/01.cir.103.10.1396.
Restenosis due to neointima formation is the major limitation of stent-supported balloon angioplasty. Despite abundant animal data, molecular mechanisms of neointima formation have been investigated on only a limited basis in patients. This study sought to establish a method for profiling gene expression in human in-stent neointima and to identify differentially expressed genes that may serve as novel therapeutic targets.
We retrieved tissue specimens from patients with symptomatic in-stent restenosis using a novel helix cutter atherectomy device. cDNA samples prepared from neointima (n=10) and, as a control, from the media of normal arteries (n=14) were amplified using a novel polymerase chain reaction protocol and hybridized to cDNA arrays. Immunohistochemistry characterized the atherectomy material as neointima. cDNA arrays readily identified differentially expressed genes. Some of the differentially expressed genes complied with expected gene expression patterns of neointima, including downregulation of desmin and upregulation of thrombospondin-1, cyclooxygenase-1, and the 70-kDa heat shock protein B. Additionally, we discovered previously unknown gene expression patterns, such as downregulation of mammary-derived growth inhibitor and upregulation of FK506-binding protein 12 (FKBP12). Upregulation of FKBP12 was confirmed at the protein level in neointimal smooth muscle cells.
Gene expression patterns of human neointima retrieved by helix-cutter atherectomy can be reliably analyzed by cDNA array technology. This technique can identify therapeutic targets in patients, as exemplified by the findings regarding FKBP12. FKBP12 is the receptor for Rapamycin (sirolimus), which in animal models reduced neointima formation. Our study thus yields a rationale for the use of Rapamycin to prevent restenosis in patients.
新生内膜形成导致的再狭窄是支架辅助球囊血管成形术的主要限制因素。尽管有大量动物实验数据，但关于新生内膜形成的分子机制在患者中仅得到有限的研究。本研究旨在建立一种分析人支架内新生内膜基因表达谱的方法，并识别可能作为新型治疗靶点的差异表达基因。
我们使用一种新型螺旋切割旋切装置从有症状的支架内再狭窄患者中获取组织标本。从新生内膜（n = 10）制备的cDNA样本，以及作为对照的正常动脉中膜（n = 14）的cDNA样本，采用一种新型聚合酶链反应方案进行扩增，并与cDNA阵列杂交。免疫组织化学将旋切材料鉴定为新生内膜。cDNA阵列能够轻松识别差异表达基因。一些差异表达基因符合新生内膜预期的基因表达模式，包括结蛋白下调以及血小板反应蛋白-1、环氧合酶-1和70 kDa热休克蛋白B上调。此外，我们还发现了以前未知的基因表达模式，如乳腺衍生生长抑制因子下调和FK506结合蛋白12（FKBP12）上调。在新生内膜平滑肌细胞的蛋白质水平证实了FKBP12上调。
通过螺旋切割旋切术获取的人新生内膜基因表达模式可通过cDNA阵列技术可靠地分析。该技术能够识别患者的治疗靶点，FKBP12的研究结果就是例证。FKBP12是雷帕霉素（西罗莫司）的受体，在动物模型中雷帕霉素可减少新生内膜形成。因此，我们的研究为使用雷帕霉素预防患者再狭窄提供了理论依据。
