Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice
Muscle overexpression of Klf15 via an AAV8-Spc5-12 construct does not provide benefits in spinal muscular atrophy mice
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通过 AAV8-Spc5-12 构建体过度表达 Klf15 对脊髓性肌萎缩症小鼠没有益处
DOI：
10.1101/717785
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Ahlskog N
中科院分区：
文献类型：
--
作者：
Ahlskog N
脊髓性肌萎缩症(SMA)是一种因存活运动神经元(SMN)基因缺失而引起的神经肌肉疾病。虽然目前有两种被批准的基于基因的治疗SMA的方法，但可用性、高昂的成本和患者反应的差异表明，需要替代治疗方案。最佳的治疗策略可能是SMN依赖和独立治疗的组合，旨在缓解中枢神经系统和外周肌肉的症状。KRüppel-like factor15(KLF15)是一种转录因子，调节肌肉中的关键代谢和麦角化途径。我们最近报道了症状前SMA小鼠肌肉中Klf15的显著下调。重要的是，围产期通过转基因和药理学方法上调Klf15基因导致SMA小鼠疾病表型的改善，包括体重和存活。在本研究中，我们设计了一个腺相关病毒血清8型(AAV8)载体，在肌肉特异性启动子Spc5-12启动子(AAV8-Klf15)下过表达密码子优化的Klf15cDNA。AAV8-Klf15对重度台湾SmN−/−、Smn2或Smn2B/−SMA小鼠肌肉中Klf15的表达显著增加。我们还观察到AAV8-Klf15载体在肝脏和心脏中具有显著的活性。AAV8介导的Klf15过表达适度提高了Smn2B/−模型的存活率，但在台湾小鼠中却没有。不能以时间和组织依赖的方式在生理水平上特异性地诱导KLF15的表达可能是导致这种有限疗效的原因之一。因此，我们的工作表明，AAV8-Spc5-12载体在包括肌肉、心脏和肝脏在内的几个组织中早在P2就能诱导高基因表达，但突出了实现载体介导的Klf15有意义转基因表达的挑战。
Spinal muscular atrophy (SMA) is a neuromuscular disease caused by loss of thesurvival motor neuron(SMN) gene. While there are currently two approved gene-based therapies for SMA, availability, high cost, and differences in patient response indicate that alternative treatment options are needed. Optimal therapeutic strategies will likely be a combination of SMN-dependent and -independent treatments aimed at alleviating symptoms in the central nervous system and peripheral muscles. Krüppel-like factor 15 (KLF15) is a transcription factor that regulates key metabolic and ergogenic pathways in muscle. We have recently reported significant downregulation ofKlf15in muscle of presymptomatic SMA mice. Importantly, perinatal upregulation ofKlf15via transgenic and pharmacological methods resulted in improved disease phenotypes in SMA mice, including weight and survival. In the current study, we designed an adeno-associated virus serotype 8 (AAV8) vector to overexpress a codon-optimizedKlf15cDNA under the muscle-specific Spc5-12 promoter (AAV8-Klf15). Administration of AAV8-Klf15to severe TaiwaneseSmn−/−;SMN2or intermediateSmn2B/−SMA mice significantly increasedKlf15expression in muscle. We also observed significant activity of the AAV8-Klf15vector in liver and heart. AAV8-mediatedKlf15overexpression moderately improved survival in theSmn2B/−model but not in the Taiwanese mice. An inability to specifically induce Klf15 expression at physiological levels in a time- and tissue-dependent manner may have contributed to this limited efficacy. Thus, our work demonstrates that an AAV8-Spc5-12 vector induces high gene expression as early as P2 in several tissues including muscle, heart, and liver, but highlights the challenges of achieving meaningful vector-mediated transgene expression of Klf15.
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影响因子：
7.7
作者：
Takashima M;Ogawa W;Hayashi K;Inoue H;Kinoshita S;Okamoto Y;Sakaue H;Wataoka Y;Emi A;Senga Y;Matsuki Y;Watanabe E;Hiramatsu R;Kasuga M
通讯作者：
Kasuga M
影响因子：
4
作者：
Boyer JG;Ferrier A;Kothary R
通讯作者：
Kothary R
影响因子：
29
作者：
Jeyaraj D;Scheer FA;Ripperger JA;Haldar SM;Lu Y;Prosdocimo DA;Eapen SJ;Eapen BL;Cui Y;Mahabeleshwar GH;Lee HG;Smith MA;Casadesus G;Mintz EM;Sun H;Wang Y;Ramsey KM;Bass J;Shea SA;Albrecht U;Jain MK
通讯作者：
Jain MK
影响因子：
46.9
作者：
Li, XY;Eastman, EM;Draghia-Akli, R
通讯作者：
Draghia-Akli, R
影响因子：
1.2
作者：
Lampe, Sara E. Gombash;Kaspar, Brian K.;Foust, Kevin D.
通讯作者：
Foust, Kevin D.