HIPPO-Integrin-linked Kinase Cross-Talk Controls Self-Sustaining Proliferation and Survival in Pulmonary Hypertension
HIPPO-Integrin-linked Kinase Cross-Talk Controls Self-Sustaining Proliferation and Survival in Pulmonary Hypertension
复制标题
DOI：
10.1164/rccm.201510-2003oc
复制
发表时间：
2016-10-01
影响因子：
24.7
通讯作者：
Goncharova, Elena A.
中科院分区：
文献类型：
--
作者：
Kudryashova, Tatiana V.;Goncharov, Dmitry A.;Goncharova, Elena A.
基本原理：肺动脉血管平滑肌细胞（PAVSMCs）增殖增强和凋亡受损是肺动脉高压（PAH）肺血管重构的关键病理生理因素。目的：探讨HIPPO信号在PAH中PAVSMCs增殖/凋亡失衡中的作用和治疗意义。使用原代远端PAVSMC、来自未使用供体（对照）和特发性PAH肺的肺组织切片以及SU 5416/低氧诱导的肺动脉高压（PH）的大鼠和小鼠模型。免疫组化，免疫细胞化学，免疫印迹分析和转染，感染，DNA合成，凋亡，迁移，细胞计数，和蛋白质活性测定进行了本study.Measurements和主要结果：免疫组化和免疫印迹分析表明，HIPPO中央组件大肿瘤抑制因子1（LATS 1）是失活的小重塑肺动脉（PA）和远端PAVSMCs在特发性PAH。基于分子和药理学的分析显示，LATS 1失活及其相互效应子Yes相关蛋白（雅普）的上调是激活哺乳动物雷帕霉素靶蛋白（mTOR）-Akt、HIF 1 α、Notch 3胞内结构域和β-连环蛋白积累、促凋亡Bim缺乏、增殖增加和人PAH PAVSMC存活所必需的。LATS 1失活和上调雅普增加了纤连蛋白的产生和分泌，从而上调了整合素连接激酶1（ILK 1）。ILK 1支持LATS 1失活，其抑制使LATS 1重新激活，下调雅普，抑制增殖，并促进PAH中的细胞凋亡，但不影响对照PAVSMC。在SU 5416/缺氧诱导PH的大鼠和小鼠的小重塑PA中，PAVSM显示LATS 1下调和ILK 1过表达。在SU 5416/缺氧暴露的第22-35天用选择性ILK抑制剂Cpd 22治疗小鼠恢复了LATS 1信号传导并减少了建立的肺血管重塑和PH。这些数据报告了HIPPO/LATS 1的失活，通过Yap-纤连蛋白-ILK 1信号环自我支持，作为PAH中PAVSMCs自我维持增殖和凋亡抵抗的新机制，并为治疗干预提供了新的潜在靶点。
Rationale: Enhanced proliferation and impaired apoptosis of pulmonary arterial vascular smooth muscle cells (PAVSMCs) are key pathophysiologic components of pulmonary vascular remodeling in pulmonary arterial hypertension (PAH).Objectives: To determine the role and therapeutic relevance of HIPPO signaling in PAVSMC proliferation/apoptosis imbalance in PAH.Methods: Primary distal PAVSMCs, lung tissue sections from unused donor (control) and idiopathic PAH lungs, and rat and mouse models of SU5416/hypoxia-induced pulmonary hypertension (PH) were used. Immunohistochemical, immunocytochemical, and immunoblot analyses and transfection, infection, DNA synthesis, apoptosis, migration, cell count, and protein activity assays were performed in this study.Measurements and Main Results: Immunohistochemical and immunoblot analyses demonstrated that the HIPPO central component large tumor suppressor 1 (LATS1) is inactivated in small remodeled pulmonary arteries (PAs) and distal PAVSMCs in idiopathic PAH. Molecular- and pharmacology-based analyses revealed that LATS1 inactivation and consequent up-regulation of its reciprocal effector Yes-associated protein (Yap) were required for activation of mammalian target of rapamycin (mTOR)-Akt, accumulation of HIFl alpha, Notch3 intracellular domain and beta-catenin, deficiency of proapoptotic Bim, increased proliferation, and survival of human PAH PAVSMCs. LATS1 inactivation and up-regulation of Yap increased production and secretion of fibronectin that up regulated integrin-linked kinase 1 (ILK1). ILK1 supported LATS1 inactivation, and its inhibition reactivated LATS1, down-regulated Yap, suppressed proliferation, and promoted apoptosis in PAH, but not control PAVSMCs. PAVSM in small remodeled PAs from rats and mice with SU5416/hypoxia-induced PH showed down regulation of LATS1 and overexpression of ILK1. Treatment of mice with selective ILKinhibitor Cpd22 at Days 22-35 of SU5416/hypoxia exposure restored LATS1 signaling and reduced established pulmonary vascular remodeling and PH.Conclusions: These data report inactivation of HIPPO/LATS1, self-supported via Yap-fibronectin-ILK1 signaling loop, as a novel mechanism of self-sustaining proliferation and apoptosis resistance of PAVSMCs in PAH and suggest a new potential target for therapeutic intervention.