A gene therapeutic approach to inhibit calcium and integrin binding protein 1 ameliorates maladaptive remodelling in pressure overload
A gene therapeutic approach to inhibit calcium and integrin binding protein 1 ameliorates maladaptive remodelling in pressure overload
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DOI：
10.1093/cvr/cvy154
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发表时间：
2019-01-01
影响因子：
10.8
通讯作者：
Heineke, Joerg
中科院分区：
文献类型：
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作者：
Grund, Andrea;Szaroszyk, Malgorzata;Heineke, Joerg
AIMS慢性心力衰竭正变得越来越普遍，并且仍然与高死亡率有关。心肌肥厚和纤维化导致心脏重构和心力衰竭，但目前的治疗策略并不能充分抑制它们。此外，尽管对心肌细胞内信号蛋白诱导病理性肥厚的了解越来越多，但目前还没有针对这些分子的治疗方法。在这项研究中，我们旨在建立和测试一种治疗工具来对抗22 kDa钙和整合素结合蛋白(CIB)1，该蛋白是病理性心肌肥厚的结节调节蛋白和非适应性钙调神经磷酸酶/NFAT轴的激活剂。方法与结果在三种不同的序列中，我们选择了一种shRNA构建物(ShCIB1)，对腺病毒过表达的新生大鼠心肌细胞(NRCM)和腺相关病毒(AAV)9载体在小鼠心脏过表达的CIB1进行了特异性的下调50%。在NRCM中过表达shCIB1显著抑制了细胞生长，改善了生物人工心脏组织的收缩性能，并降低了对肥大刺激的钙调神经磷酸酶/NFAT的激活。在小鼠中，注射AAV-shCIB1可显著改善由横动脉缩窄(TAC)引起的压力超负荷2周期间的离心性心肌肥厚和心功能障碍。超微结构和分子分析显示，与AAV-shControl相比，AAV-shCIB1组小鼠TAC后心肌纤维化明显减轻，肥大相关基因表达和钙调神经磷酸酶/NFAT以及ERK MAPK活性显著降低。在长期压力超负荷暴露10周期间，AAV-shCIB1治疗维持其抗肥厚和抗纤维化的作用，但心功能不再改善，这很可能是由于CIB1表达下调导致心肌血管生成减少所致。结论shRNA介导的抑制CIB1基因治疗有效地抑制了压力超负荷时的病理性心肌肥大和纤维化。虽然shCIB1最初可以改善心脏功能，但在持续超负荷期间，这种改善无法保持。
Aims Chronic heart failure is becoming increasingly prevalent and is still associated with a high mortality rate. Myocardial hypertrophy and fibrosis drive cardiac remodelling and heart failure, but they are not sufficiently inhibited by current treatment strategies. Furthermore, despite increasing knowledge on cardiomyocyte intracellular signalling proteins inducing pathological hypertrophy, therapeutic approaches to target these molecules are currently unavailable. In this study, we aimed to establish and test a therapeutic tool to counteract the 22kDa calcium and integrin binding protein (CIB) 1, which we have previously identified as nodal regulator of pathological cardiac hypertrophy and as activator of the maladaptive calcineurin/NFAT axis.Methods and results Among three different sequences, we selected a shRNA construct (shCIB1) to specifically down-regulate CIB1 by 50% upon adenoviral overexpression in neonatal rat cardiomyocytes (NRCM), and upon overexpression by an adeno-associated-virus (AAV) 9 vector in mouse hearts. Overexpression of shCIB1 in NRCM markedly reduced cellular growth, improved contractility of bioartificial cardiac tissue and reduced calcineurin/NFAT activation in response to hypertrophic stimulation. In mice, administration of AAV-shCIB1 strongly ameliorated eccentric cardiac hypertrophy and cardiac dysfunction during 2 weeks of pressure overload by transverse aortic constriction (TAC). Ultrastructural and molecular analyses revealed markedly reduced myocardial fibrosis, inhibition of hypertrophy associated gene expression and calcineurin/NFAT as well as ERK MAP kinase activation after TAC in AAV-shCIB1 vs. AAV-shControl treated mice. During long-term exposure to pressure overload for 10weeks, AAV-shCIB1 treatment maintained its anti-hypertrophic and anti-fibrotic effects, but cardiac function was no longer improved vs. AAV-shControl treatment, most likely resulting from a reduction in myocardial angiogenesis upon downregulation of CIB1.Conclusions Inhibition of CIB1 by a shRNA-mediated gene therapy potently inhibits pathological cardiac hypertrophy and fibrosis during pressure overload. While cardiac function is initially improved by shCIB1, this cannot be kept up during persisting overload.