Heart failure-inducible gene therapy targeting protein phosphatase 1 prevents progressive left ventricular remodeling.
Heart failure-inducible gene therapy targeting protein phosphatase 1 prevents progressive left ventricular remodeling.
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DOI：
10.1371/journal.pone.0035875
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Matsuzaki M
中科院分区：
文献类型：
--
作者：
Miyazaki Y;Ikeda Y;Shiraishi K;Fujimoto SN;Aoyama H;Yoshimura K;Inui M;Hoshijima M;Kasahara H;Aoki H;Matsuzaki M
靶向钙循环已成为治疗严重心力衰竭的一种潜在疗法。这些方法包括针对肌浆网(SR)Ca~(2+)ATPase过表达的基因治疗，或去除磷蛋白(PLN)和相关蛋白磷酸酶1(PP1)蛋白复合体。我们先前报道，Pp1β是Pp1催化亚基之一，在三种Pp1亚型中，它主要抑制SR的钙摄取，从而有助于心衰时钙的下调。在本研究中，我们研究了用腺相关病毒-9(AAV9)载体介导的基因治疗来抑制心力衰竭诱导的Pp1β是否有利于防止遗传性心肌病小鼠的疾病进展。我们构建了编码PP1shRNA或阴性对照β的腺相关病毒载体AAV9。利用与翡翠绿荧光蛋白(EmGFP)连接的B型利钠蛋白(BNP)启动子和shRNA序列构建心力衰竭诱导基因表达系统。将AAV9载体(AAV9-BNP-EmGFP-PP1βshRNA和AAV9-BNP-EmGFP-NCshRNA)注入肌肉LIM蛋白缺陷小鼠尾静脉(2×1011GC/只)，3个月后进行超声心动图、血流动力学、生化和组织学分析。在MLPKO小鼠中，通过检测EmGFP的表达和诱导心肌中pp1β的减少25%，显示出BNP启动子的活性增加。可诱导的PP1shRNA递送优先改善左室舒张功能和减轻不良的室壁重构。与AAV9-BNP-EmGFP-NCshRNA组相比，AAV9-BNP-EmGFP-PP1βshRNA组PLN磷酸化明显增强。此外，BNP的产生在3个月时减少，心脏间质纤维化被消除。心力衰竭诱导的分子靶向Pp1β具有潜在的治疗心力衰竭的新策略。
The targeting of Ca2+ cycling has emerged as a potential therapy for the treatment of severe heart failure. These approaches include gene therapy directed at overexpressing sarcoplasmic reticulum (SR) Ca2+ ATPase, or ablation of phospholamban (PLN) and associated protein phosphatase 1 (PP1) protein complexes. We previously reported that PP1β, one of the PP1 catalytic subunits, predominantly suppresses Ca2+ uptake in the SR among the three PP1 isoforms, thereby contributing to Ca2+ downregulation in failing hearts. In the present study, we investigated whether heart-failure-inducible PP1β-inhibition by adeno-associated viral-9 (AAV9) vector mediated gene therapy is beneficial for preventing disease progression in genetic cardiomyopathic mice. We created an adeno-associated virus 9 (AAV9) vector encoding PP1β short-hairpin RNA (shRNA) or negative control (NC) shRNA. A heart failure inducible gene expression system was employed using the B-type natriuretic protein (BNP) promoter conjugated to emerald-green fluorescence protein (EmGFP) and the shRNA sequence. AAV9 vectors (AAV9-BNP-EmGFP-PP1βshRNA and AAV9-BNP-EmGFP-NCshRNA) were injected into the tail vein (2×1011 GC/mouse) of muscle LIM protein deficient mice (MLPKO), followed by serial analysis of echocardiography, hemodynamic measurement, biochemical and histological analysis at 3 months. In the MLPKO mice, BNP promoter activity was shown to be increased by detecting both EmGFP expression and the induced reduction of PP1β by 25% in the myocardium. Inducible PP1βshRNA delivery preferentially ameliorated left ventricular diastolic function and mitigated adverse ventricular remodeling. PLN phosphorylation was significantly augmented in the AAV9-BNP-EmGFP-PP1βshRNA injected hearts compared with the AAV9-BNP-EmGFP-NCshRNA group. Furthermore, BNP production was reduced, and cardiac interstitial fibrosis was abrogated at 3 months. Heart failure-inducible molecular targeting of PP1β has potential as a novel therapeutic strategy for heart failure.
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