Effect of vectors on human endothelial cell signal transduction: implications for cardiovascular gene therapy.
Effect of vectors on human endothelial cell signal transduction: implications for cardiovascular gene therapy.
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载体对人内皮细胞信号转导的影响：对心血管基因治疗的影响。
DOI：
10.1016/j.ymthe.2006.08.1087
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
A. George
中科院分区：
文献类型：
--
作者：
P. Tan;S. Xue;M. Manunta;S. Beutelspacher;H. Fazekasova;A. Alam;M. Mcclure;A. George
目的：内皮素是基因治疗的重要靶点。我们已经研究了病毒和非病毒载体对内皮细胞（ECs）的表型和功能的影响，并开发了方法来阻止任何激活引起的这些vectors.Methods和结果：转导的ECs与病毒载体，包括腺病毒，慢病毒和莫洛尼鼠白血病病毒可以诱导促炎表型。我们发现，病毒载体导致相当大的上调粘附分子和MHC II分子。我们看到了上调的Th 1细胞因子，以及增强分泌的促炎细胞因子在EC病毒转导后。抑制NFκB不能完全消除病毒诱导的粘附分子表达和促炎细胞因子的产生。然而，PI 3-K和PKR的抑制导致EC激活的完全抑制。这表明PI 3-K和PKR通路的激活不仅激活了NFκB通路，而且还激活了其他下游效应通路。使用其他抑制剂表明阻断MAPK p38、JNK、STAT-1和STAT 3通路对病毒转导激活EC的影响很小。PKR和PI 3-K通路均与细胞因子产生（PKR的IFN-α/β和PI 3-K的TNF-α/IL-1α/β）相关，其作为这些通路激活的正反馈环。这些可能有助于将EC“锁定”为活化的表型。为了测试EC的病毒转导的功能性后果，我们将EC与病毒载体一起孵育，然后将它们用于粒细胞、淋巴细胞和单核细胞的粘附和迁移测定。病毒载体诱导的粘附作用与细胞因子激活EC后观察到的粘附作用相当。病毒对EC的转导作用可以变化。我们还证明，病毒激活的EC未能响应随后激活的促炎细胞因子。我们发现，长期刺激的促炎细胞因子的EC未能上调促炎表型后，随后的激活，以类似的方式，以病毒转导的EC。如果中和抗体，这些细胞因子被添加从病毒转导的时间，内皮细胞能够响应激活刺激以类似的方式未transduced ECs.Conclusions：我们已经表明，与病毒载体的转导有显着的后果内皮细胞在他们的表型，激活途径诱导和他们的功能。这可以通过诱导局部炎症来调节基因治疗的效果，这两者都可以调节潜在的疾病过程。另一方面，非病毒载体相对无害，不能显著激活EC。然而，它们在介导基因表达方面效率较低！了解EC转导的后果及其介导的途径，对于设计涉及EC病毒载体修饰的改进策略非常重要。在这方面，我们已经表明，阻断参与病毒活化的信号通路可用于防止这种活化
Objective:Endothelium is an important target for gene therapy. We have investigated the effect of viral and non-viral vectors on the phenotype and function of endothelial cells (ECs), and developed methods to block any activation caused by these vectors.Methods and results:Transduction of ECs with viral vectors, including adenovirus, lentiviruses and Moloney murine leukemia virus can induce a pro-inflammatory phenotype. We showed that viral vectors resulted in considerable upregulation of adhesion molecules and MHC II molecules. We saw an upregulation of Th1 cytokines, as well as enhanced secretion of proinflammatory cytokines in EC following viral transductions. Inhibition of NFκB did not completely abolish virally induced adhesion molecule expression and pro-inflammatory cytokine production. However, inhibition of both PI3-K and PKR resulted in total inhibition of EC activation. This suggests that activation of PI3-K and PKR pathways not only activate NFκB pathway, but also other downstream effector pathways. Use of other inhibitors suggested that blockade of the MAPK p38, JNK, STAT-1 and STAT3 pathways had little effect on the activation of ECs by viral transduction. Both the PKR and the PI3-K pathways are associated with cytokine production (IFN-α/β for PKR and TNF-α/IL-1α/β for PI3-K) that serves as a positive feedback loop for the activation of those pathways. These might serve to help ‘lock’the EC into an activated phenotype. In order to test the functional consequences of viral transduction of EC, we incubated EC with viral vectors then used them as in adhesion and transmigration assays with granulocytes, lymphocytes and monocytes. Viral vector-induced adhesive effect was comparable to that seen following cytokine activation of the ECs. Viral transduction effect on EC can be varied. We also demonstrated that virally activated ECs failed to respond to subsequent activation by proinflammatory cytokines. We showed that EC that were chronically stimulated with pro-inflammatory cytokines failed to upregulate pro-inflammatory phenotypes upon subsequent activation, in a similar manner to virally transduced ECs. If neutralising antibodies to these cytokines were added from the time of viral transduction, the ECs were able to respond to activation stimuli in a similar manner to untransduced ECs.Conclusions:We have shown that transduction with viral vectors has dramatic consequences for ECs both in terms of their phenotype, the activation pathways induced and their function. This can modulate the effect of gene therapy by inducing local inflammation, which may both modulate the underlying disease process. Non-viral vectors, on the other hand, are relatively innocuous and fail to activate significantly ECs. However, they are less efficient at mediating gene expression! An understanding of both the consequences of EC transduction, and the pathways by which it is mediated, is important in allowing the design of improved strategies involving the viral vector modification of ECs. In this regard we have shown that blockade of the signaling pathways involved in viral activation may be used to prevent such activation
通过逆转录病毒介导的前列腺素 H 合酶 cDNA 转移增强内皮细胞中的前列环素合成。
DOI：
10.1172/jci116398
发表时间：
1993
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Xu,XM;Ohashi,K;Sanduja,SK;Ruan,KH;Wang,LH;Wu,KK
通讯作者：
Wu,KK
用重组腺病毒载体感染 A549 细胞可诱导 ICAM-1 表达并增加活化中性粒细胞的 CD-18 依赖性粘附。
影响因子：
4.2
作者：
Stark,JM;Amin,RS;Trapnell,BC
通讯作者：
Trapnell,BC
DOI：
10.1073/pnas.0308368101
发表时间：
2004-04-20
影响因子：
11.1
作者：
Philpott, NJ;Nociari, M;Falck-Pedersen, E
通讯作者：
Falck-Pedersen, E