Genome-wide identification of FoxO-dependent gene networks in skeletal muscle during C26 cancer cachexia.
Genome-wide identification of FoxO-dependent gene networks in skeletal muscle during C26 cancer cachexia.
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DOI：
10.1186/1471-2407-14-997
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发表时间：
2014-12-24
期刊：
影响因子：
3.8
通讯作者：
Judge AR
中科院分区：
文献类型：
--
作者：
Judge SM;Wu CL;Beharry AW;Roberts BM;Ferreira LF;Kandarian SC;Judge AR
来自恶病质癌症患者和癌症恶病质动物模型的证据支持叉头盒O(FoxO)转录因子参与驱动癌症诱导的骨骼肌萎缩。然而，FoxO在癌症恶病质过程中调控的全基因组基因网络和相关的生物学过程尚不清楚。我们假设FoxO是癌症期间骨骼肌不同基因网络的中心上游调节因子，可能协同作用促进消瘦表型。为了抑制内源性FoxO DNA结合，我们用含有显性阴性(D.N.)的AAV9的AAV9基因转导小鼠的四肢和横隔肌。FOXO蛋白(或GFP对照)。D.n.FoxO结构仅由FOXO3a DNA结合域组成，该结构域与FoxO1和FOXO4高度同源，并竞争并阻止内源性FoxO DNA结合。随后将26天后收获的结肠-26(C26)细胞和肌肉接种给小鼠。阻断FoxO可阻止C26引起的运动肌和横隔肌的肌纤维萎缩，并显著避免力量缺陷。肌肉大小和功能的这种保留与对C26反应改变的543个转录本(2093个)的差异调节有关。对需要FoxO的上调基因转录本的生物信息学分析显示，蛋白酶体、AP-1和IL-6途径丰富，并包括几个与萎缩相关的转录因子，包括STAT3、Fos和CEBPB。FOXO对于癌症诱导的几个基因转录本的下调也是必要的，这些基因转录本富含细胞外基质和肌节蛋白编码基因。我们通过qRT-PCR法在四肢肌肉和横隔肌中验证了这些发现，并进一步证明FOXO1和/或FOXO3a足以增加STAT3、Fos、CEBPB和C/EBPβ靶基因Ubr2。对CEBPB近端启动子的分析揭示了两个真实的FoxO结合元件，我们进一步证实，这两个元件对于CEBPB启动子对IL-6的激活是必需的，IL-6是C26肿瘤模型中的主要细胞因子。这些发现提供了新的证据，证明在癌症恶病质过程中，FoxO依赖的转录是控制骨骼肌不同基因网络的中心节点，并发现了新的候选基因和网络，作为癌症诱导的消瘦的致病因素，有待进一步研究。本文的在线版本(DOI：10.1186/1471-2407-14-997)包含补充材料，可供授权用户使用。
Evidence from cachectic cancer patients and animal models of cancer cachexia supports the involvement of Forkhead box O (FoxO) transcription factors in driving cancer-induced skeletal muscle wasting. However, the genome-wide gene networks and associated biological processes regulated by FoxO during cancer cachexia are unknown. We hypothesize that FoxO is a central upstream regulator of diverse gene networks in skeletal muscle during cancer that may act coordinately to promote the wasting phenotype. To inhibit endogenous FoxO DNA-binding, we transduced limb and diaphragm muscles of mice with AAV9 containing the cDNA for a dominant negative (d.n.) FoxO protein (or GFP control). The d.n.FoxO construct consists of only the FoxO3a DNA-binding domain that is highly homologous to that of FoxO1 and FoxO4, and which outcompetes and blocks endogenous FoxO DNA binding. Mice were subsequently inoculated with Colon-26 (C26) cells and muscles harvested 26 days later. Blocking FoxO prevented C26-induced muscle fiber atrophy of both locomotor muscles and the diaphragm and significantly spared force deficits. This sparing of muscle size and function was associated with the differential regulation of 543 transcripts (out of 2,093) which changed in response to C26. Bioinformatics analysis of upregulated gene transcripts that required FoxO revealed enrichment of the proteasome, AP-1 and IL-6 pathways, and included several atrophy-related transcription factors, including Stat3, Fos, and Cebpb. FoxO was also necessary for the cancer-induced downregulation of several gene transcripts that were enriched for extracellular matrix and sarcomere protein-encoding genes. We validated these findings in limb muscles and the diaphragm through qRT-PCR, and further demonstrate that FoxO1 and/or FoxO3a are sufficient to increase Stat3, Fos, Cebpb, and the C/EBPβ target gene, Ubr2. Analysis of the Cebpb proximal promoter revealed two bona fide FoxO binding elements, which we further establish are necessary for Cebpb promoter activation in response to IL-6, a predominant cytokine in the C26 cancer model. These findings provide new evidence that FoxO-dependent transcription is a central node controlling diverse gene networks in skeletal muscle during cancer cachexia, and identifies novel candidate genes and networks for further investigation as causative factors in cancer-induced wasting. The online version of this article (doi:10.1186/1471-2407-14-997) contains supplementary material, which is available to authorized users.
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DOI：
10.1016/j.bbrc.2008.11.043
发表时间：
2009-01-09
影响因子：
3.1
作者：
Ito, Yu;Daitoku, Hiroaki;Fukamizu, Akiyoshi
通讯作者：
Fukamizu, Akiyoshi
影响因子：
4.8
作者：
Lecker, SH;Jagoe, RT;Goldberg, AL
通讯作者：
Goldberg, AL
影响因子：
3.2
作者：
Bachmann, Jeannine;Heiligensetzer, Mathias;Martignoni, Marc E.
通讯作者：
Martignoni, Marc E.
影响因子：
4.8
作者：
Birkenkamp, Kim U.;Essafi, Abdelkader;Coffer, Paul J.
通讯作者：
Coffer, Paul J.
影响因子：
50.3
作者：
Acharyya, S;Butchbach, MER;Guttridge, DC
通讯作者：
Guttridge, DC