A brain-sparing diphtheria toxin for chemical genetic ablation of peripheral cell lineages
A brain-sparing diphtheria toxin for chemical genetic ablation of peripheral cell lineages
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DOI：
10.1038/ncomms14967
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发表时间：
2017-04-03
影响因子：
16.6
通讯作者：
Domingos, Ana I.
中科院分区：
文献类型：
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作者：
Pereira, Mafalda M. A.;Mahu, Ines;Domingos, Ana I.
白喉毒素受体（DTR）的条件表达被广泛用于组织特异性细胞消融。然而，白喉毒素（DT）穿过血脑屏障，这限制了它使用同样在中枢神经系统（CNS）中表达的Cre驱动因子消融外周细胞的效用。在这里，我们报告了脑保留DT的发展，称为BRAINSPAReDT，用于中枢神经系统外细胞的组织特异性基因消融。我们通过PEGylation阻止DT通过血脑屏障，使分子极化并增加其大小。我们通过区域遗传交感神经切除术验证了BRAINSPAReDT: BRAINSPAReDT切除外周而非中枢儿茶酚胺能神经元，从而避免了与多巴胺能完全耗尽相关的帕金森样表型。局部交感神经切除术损害脂肪组织产热，使小鼠易患肥胖。我们提供了一个原理证明，BRAINSPAReDT可以用于CNS驱动的脑外Cre/DTR组织特异性消融，同时巩固肥胖与交感神经系统之间的联系。
Conditional expression of diphtheria toxin receptor (DTR) is widely used for tissue-specific ablation of cells. However, diphtheria toxin (DT) crosses the blood-brain barrier, which limits its utility for ablating peripheral cells using Cre drivers that are also expressed in the central nervous system (CNS). Here we report the development of a brain-sparing DT, termed BRAINSPAReDT, for tissue-specific genetic ablation of cells outside the CNS. We prevent blood-brain barrier passage of DT through PEGylation, which polarizes the molecule and increases its size. We validate BRAINSPAReDT with regional genetic sympathectomy: BRAINSPAReDT ablates peripheral but not central catecholaminergic neurons, thus avoiding the Parkinson-like phenotype associated with full dopaminergic depletion. Regional sympathectomy compromises adipose tissue thermogenesis, and renders mice susceptible to obesity. We provide a proof of principle that BRAINSPAReDT can be used for Cre/DTR tissue-specific ablation outside the brain using CNS drivers, while consolidating the link between adiposity and the sympathetic nervous system.