Gene therapy by targeted adenovirus-mediated knockdown of pulmonary endothelial Tph1 attenuates hypoxia-induced pulmonary hypertension.
Gene therapy by targeted adenovirus-mediated knockdown of pulmonary endothelial Tph1 attenuates hypoxia-induced pulmonary hypertension.
复制标题
DOI：
10.1038/mt.2012.70
复制
发表时间：
2012-08
期刊：
影响因子：
--
通讯作者：
I. Morecroft;K. White;P. Caruso;M. Nilsen;Lynn Loughlin;R. Alba;P. Reynolds;S. Danilov;A. Baker-A
中科院分区：
文献类型：
--
作者：
I. Morecroft;K. White;P. Caruso;M. Nilsen;Lynn Loughlin;R. Alba;P. Reynolds;S. Danilov;A. Baker-A
血清素是由肺动脉内皮细胞（PAEC）通过色氨酸羟化酶-1 （Tph1）产生的。病理上，血清素作用于潜在的肺动脉细胞，促进与肺动脉高压（PAH）相关的血管重塑。缺氧对PAEC-Tph1活性的影响尚不清楚。我们研究了在PAH缺氧模型中选择性抑制paec - tph1基因治疗PAH方法的潜力。我们在缺氧暴露前后将培养的牛肺动脉平滑肌细胞（bPASMCs）暴露于人肺动脉平滑肌细胞（hPAECs）的条件培养基中。在缺氧PAEC培养基中血清素水平升高。条件培养基通过血清素依赖机制诱导bPASMC增殖，缺氧PAEC培养基诱导的bPASMC增殖更大。在体内，使用靶向PAECs的腺病毒载体（利用血管紧张素转换酶（ACE）双特异性抗体作为选择性靶向系统）在大鼠体内传递小发夹Tph1 RNA序列。缺氧大鼠出现多环芳烃，肺Tph1升高。我们的PAEC-Tph1基因敲除策略可减弱PAEC-Tph1的表达和PAH的发展。这些结果表明，缺氧可诱导Tph1活性，paec -Tph1的选择性下调可减轻大鼠缺氧诱导的PAH。通过基因干预进一步研究肺内皮特异性Tph1抑制是必要的。
Serotonin is produced by pulmonary arterial endothelial cells (PAEC) via tryptophan hydroxylase-1 (Tph1). Pathologically, serotonin acts on underlying pulmonary arterial cells, contributing to vascular remodeling associated with pulmonary arterial hypertension (PAH). The effects of hypoxia on PAEC-Tph1 activity are unknown. We investigated the potential of a gene therapy approach to PAH using selective inhibition of PAEC-Tph1in vivoin a hypoxic model of PAH. We exposed cultured bovine pulmonary arterial smooth muscle cells (bPASMCs) to conditioned media from human PAECs (hPAECs) before and after hypoxic exposure. Serotonin levels were increased in hypoxic PAEC media. Conditioned media evoked bPASMC proliferation, which was greater with hypoxic PAEC media, via a serotonin-dependent mechanism.In vivo, adenoviral vectors targeted to PAECs (utilizing bispecific antibody to angiotensin-converting enzyme (ACE) as the selective targeting system) were used to deliver small hairpin Tph1 RNA sequences in rats. Hypoxic rats developed PAH and increased lung Tph1. PAEC-Tph1 expression and development of PAH were attenuated by our PAEC-Tph1gene knockdown strategy. These results demonstrate that hypoxia induces Tph1 activity and selective knockdown of PAEC-Tph1attenuates hypoxia-induced PAH in rats. Further investigation of pulmonary endothelial-specific Tph1 inhibition via gene interventions is warranted.