Use of genetically encoded, light-gated ion translocators to control tumorigenesis.
Use of genetically encoded, light-gated ion translocators to control tumorigenesis.
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DOI：
10.18632/oncotarget.8036
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发表时间：
2016-04-12
期刊：
影响因子：
--
通讯作者：
Levin M
中科院分区：
文献类型：
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作者：
Chernet BT;Adams DS;Lobikin M;Levin M
人们早就知道肿瘤细胞的静息电位相对于正常细胞是去极化的。最近的工作提供的证据表明，静息电位不仅仅是细胞状态的读数：它还调节细胞行为。因此，控制体内静息电位的能力将为肿瘤的研究和治疗提供强大的新工具，该工具能够揭示活态生理信息，而使用应用于分离细胞成分的分子工具无法获得这些信息。在这里，我们描述了首次使用光遗传学来操纵离子流介导的膜电位调节，特别是为了预防和引起癌基因诱导的肿瘤的消退。在非洲爪蟾蝌蚪中，注射突变型 KRAS mRNA 可诱导肿瘤样结构，其与肿瘤有许多已记录的相似之处。我们发现，ChR2D156A（蓝光激活的阳离子通道）或 Arch（绿光激活的质子泵）的表达和激活（两者均使细胞超极化）可显着降低 KRAS 肿瘤形成的发生率。令人兴奋的是，我们还证明，肿瘤形成后共表达的光激活离子易位子的激活显着增加了肿瘤在正常化过程中消退的频率。这些数据证明了一种光遗传学方法可以剖析癌症的生物物理学。此外，它们为一类新型干预措施提供了原理验证，这些干预措施旨在通过针对可以克服突变存在的生理调节因子来调节细胞状态。
It has long been known that the resting potential of tumor cells is depolarized relative to their normal counterparts. More recent work has provided evidence that resting potential is not just a readout of cell state: it regulates cell behavior as well. Thus, the ability to control resting potential in vivo would provide a powerful new tool for the study and treatment of tumors, a tool capable of revealing living-state physiological information impossible to obtain using molecular tools applied to isolated cell components. Here we describe the first use of optogenetics to manipulate ion-flux mediated regulation of membrane potential specifically to prevent and cause regression of oncogene-induced tumors. Injection of mutant-KRAS mRNA induces tumor-like structures with many documented similarities to tumors, in Xenopus tadpoles. We show that expression and activation of either ChR2D156A, a blue-light activated cation channel, or Arch, a green-light activated proton pump, both of which hyperpolarize cells, significantly lowers the incidence of KRAS tumor formation. Excitingly, we also demonstrate that activation of co-expressed light-activated ion translocators after tumor formation significantly increases the frequency with which the tumors regress in a process called normalization. These data demonstrate an optogenetic approach to dissect the biophysics of cancer. Moreover, they provide proof-of-principle for a novel class of interventions, directed at regulating cell state by targeting physiological regulators that can over-ride the presence of mutations.