Control of very low-density lipoprotein secretion by N-ethylmaleimide-sensitive factor and miR-33.
Control of very low-density lipoprotein secretion by N-ethylmaleimide-sensitive factor and miR-33.
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DOI：
10.1161/circresaha.115.303100
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发表时间：
2014-06-20
影响因子：
20.1
通讯作者：
Baldán A
中科院分区：
文献类型：
--
作者：
Allen RM;Marquart TJ;Jesse JJ;Baldán A
一些报告表明，针对miR-33的反义寡核苷酸可能通过加速胆固醇逆向转运途径来降低患者的心血管风险。然而，关于抗miR-33治疗对极低密度脂蛋白-甘油三酯（VLDL-TAG）水平的影响，存在相互矛盾的报告。我们检验了miR-33控制肝脏VLDL-TAG分泌的假设。使用miR-33的治疗性沉默和miR-33的腺病毒过表达，我们表明miR-33通过靶向N-乙基马来酰亚胺敏感因子（NSF）限制VLDL-TAG的肝脏分泌，在体内和原代肝细胞中均如此。我们鉴定了NSF的3'UTR中的保守序列作为miR-33响应元件，并表明在miR-33诱导后，Nsf被特异性地募集到RNA诱导的沉默复合物（RISC）中。在脉冲追踪实验中，与对照细胞相比，miR-33过表达或Nsf敲低导致原代肝细胞中脱辅基蛋白和TAG的分泌减少。重要的是，Nsf挽救了miR-33依赖性的分泌减少。最后，我们表明，在体内过表达的Nsf增加全球肝脏分泌和提高血浆VLDL-TAG。总之，我们的数据揭示了miR-33-NSF轴在肝脏分泌过程中的关键作用，并表明应谨慎使用基于抗miR-33的治疗，因为它们可能会提高促动脉粥样硬化的VLDL-TAG水平。
Several reports suggest that antisense oligonucleotides against miR-33 might reduce cardiovascular risk in patients by accelerating the reverse cholesterol transport pathway. However, conflicting reports exist regarding the impact of anti-miR-33 therapy on the levels of very low-density lipoprotein-triglycerides (VLDL-TAG). We test the hypothesis that miR-33 controls hepatic VLDL-TAG secretion. Using therapeutic silencing of miR-33 and adenoviral overexpression of miR-33, we show that miR-33 limits hepatic secretion of VLDL-TAG by targeting N-ethylmaleimide-sensitive factor (NSF), both in vivo and in primary hepatocytes. We identify conserved sequences in the 3’UTR of NSF as miR-33 responsive elements, and show that Nsf is specifically recruited to the RNA-Induced Silencing Complex (RISC) following induction of miR-33. In pulse-chase experiments, either miR-33 overexpression or knock-down of Nsf lead to decreased secretion of apoproteins and TAG in primary hepatocytes, compared to control cells. Importantly, Nsf rescues miR-33–dependent reduced secretion. Finally, we show that overexpression of Nsf in vivo increases global hepatic secretion and raises plasma VLDL-TAG. Together, our data reveal key roles for the miR-33–NSF axis during hepatic secretion, and suggest that caution should be taken with anti-miR-33–based therapies since they might raise pro-atherogenic VLDL-TAG levels.