Tissue-restricted inhibition of mTOR using chemical genetics.
Tissue-restricted inhibition of mTOR using chemical genetics.
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DOI：
10.1073/pnas.2204083119
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发表时间：
2022-09-20
影响因子：
11.1
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中科院分区：
文献类型：
--
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哺乳动物雷帕霉素靶点（mTOR）在包括细胞生长、发育、免疫和衰老在内的许多重要生物体中起着重要作用，但解剖mTOR的组织特异性影响已被证明具有挑战性。这项工作描述了一个简单的系统，用于识别负责mTOR不同功能的特定组织和细胞，我们表明我们的系统可以用于从酵母到人类的生物。哺乳动物雷帕霉素靶蛋白（mTOR）是一种高度保守的真核蛋白激酶，协调细胞生长和代谢，在癌症、免疫和衰老中起关键作用。目前尚不清楚单个组织中的mTOR信号传导如何促进整个生物体的过程，因为mTOR抑制剂，如天然产物雷帕霉素，是全身给药的，同时靶向多个组织。我们开发了一种化学遗传系统，称为选择器，通过靶向表达突变体FKBP12蛋白来限制雷帕霉素类似物对特定细胞群的活性。该类似物降低了其专性结合伙伴FKBP12的亲和力，从而降低了其在野生型细胞和组织中抑制mTOR的能力。突变体FKBP12的表达，包含一个扩大的结合袋，恢复了这种雷帕霉素类似物的活性。使用该系统，我们证明了选择性mTOR抑制可以在酿酒酵母和人类细胞中实现，并且我们通过识别负责雷帕霉素诱导的果蝇发育延迟的组织，验证了我们的系统在完整的后生动物模型生物中的效用。
Mammalian target of rapamycin (mTOR) plays a number of critical organismal roles, including in cell growth, development, immunity, and aging, but dissecting the tissue-specific influences of mTOR has proven challenging. This work describes a simple system for identifying the specific tissues and cells responsible for the diverse functions of mTOR, and we show that our system can be used in organisms ranging from yeast to humans. Mammalian target of rapamycin (mTOR) is a highly conserved eukaryotic protein kinase that coordinates cell growth and metabolism, and plays a critical role in cancer, immunity, and aging. It remains unclear how mTOR signaling in individual tissues contributes to whole-organism processes because mTOR inhibitors, like the natural product rapamycin, are administered systemically and target multiple tissues simultaneously. We developed a chemical-genetic system, termed selecTOR, that restricts the activity of a rapamycin analog to specific cell populations through targeted expression of a mutant FKBP12 protein. This analog has reduced affinity for its obligate binding partner FKBP12, which reduces its ability to inhibit mTOR in wild-type cells and tissues. Expression of the mutant FKBP12, which contains an expanded binding pocket, rescues the activity of this rapamycin analog. Using this system, we show that selective mTOR inhibition can be achieved in Saccharomyces cerevisiae and human cells, and we validate the utility of our system in an intact metazoan model organism by identifying the tissues responsible for a rapamycin-induced developmental delay in Drosophila.
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影响因子：
7.7
作者：
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通讯作者：
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作者：
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