Non-invasive monitoring of alternative splicing outcomes to identify candidate therapies for myotonic dystrophy type 1
Non-invasive monitoring of alternative splicing outcomes to identify candidate therapies for myotonic dystrophy type 1
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DOI：
10.1038/s41467-018-07517-y
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发表时间：
2018-12-07
影响因子：
16.6
通讯作者：
Wheeler, Thurman M.
中科院分区：
文献类型：
--
作者：
Hu, Ningyan;Antoury, Layal;Wheeler, Thurman M.
在药物开发过程中，组织样本作为疾病活动性和药效学反应的指标。可靠的非侵入性药物靶标接触测量将有助于确定有希望的新治疗方法。在这里，我们开发并验证了一种新的双转基因1型肌强直性营养不良（DM1）小鼠模型，其中DsRed或GFP的表达是由DM1中失调的上游小基因的选择性剪接决定的。使用一种新的体内荧光光谱系统，我们发现定量DsRed/GFP比率可以准确估计活体小鼠肌肉组织中的剪接结果，其吞吐量几乎是传统荧光成像技术的两倍。用C16脂肪酸配体偶联反义（LICA）寡核苷酸治疗小鼠的一系列体内光谱测量显示，在7天内出现剂量依赖性治疗反应，证实了持续数周的作用，并证明与未偶联的亲本寡核苷酸相比，靶向性提高了两倍。
During drug development, tissue samples serve as indicators of disease activity and pharmacodynamic responses. Reliable non-invasive measures of drug target engagement will facilitate identification of promising new treatments. Here we develop and validate a novel bi-transgenic mouse model of myotonic dystrophy type 1 (DM1) in which expression of either DsRed or GFP is determined by alternative splicing of an upstream minigene that is misregulated in DM1. Using a novel in vivo fluorescence spectroscopy system, we show that quantitation of the DsRed/GFP ratio provides an accurate estimation of splicing outcomes in muscle tissue of live mice that nearly doubles throughput over conventional fluorescence imaging techniques. Serial in vivo spectroscopy measurements in mice treated with a C16 fatty acid ligand conjugated antisense (LICA) oligonucleotide reveal a dose-dependent therapeutic response within seven days, confirm a several-week duration of action, and demonstrate a two-fold greater target engagement as compared to the unconjugated parent oligonucleotide.