RNAi screening in primary human hepatocytes of genes implicated in genome-wide association studies for roles in type 2 diabetes identifies roles for CAMK1D and CDKAL1, among others, in hepatic glucose regulation.
RNAi screening in primary human hepatocytes of genes implicated in genome-wide association studies for roles in type 2 diabetes identifies roles for CAMK1D and CDKAL1, among others, in hepatic glucose regulation.
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DOI：
10.1371/journal.pone.0064946
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Tien E
中科院分区：
文献类型：
--
作者：
Haney S;Zhao J;Tiwari S;Eng K;Guey LT;Tien E
全基因组关联（GWA）研究已经描述了大量导致2型糖尿病（T2 D）的新候选基因。在某些情况下，涉及小的基因簇，而不是单个基因，并且在所有情况下，遗传贡献不是通过对特定器官（如胰腺或肝脏）的影响来定义的。有一个显着的需要，开发和使用人类细胞为基础的模型，以检查这些基因可能对葡萄糖调节的影响。我们描述了一个原代人肝细胞模型，根据激素信号调节葡萄糖处置的发展。该模型用于确定在GWA研究中鉴定的候选基因是否通过与鉴定的基因列表对应的siRNA调节肝葡萄糖处置。我们发现有几个基因影响葡萄糖作为糖原的储存（糖酵解反应）和/或影响丙酮酸的利用，这是糖原合成的关键步骤。在影响这两个过程的基因中，CAMK 1D、TSPAN 8和KIF 11影响胰高血糖素应答时，将胰高血糖素生成和糖酵解调节的介质CRTC 2定位到细胞核。此外，观察到基因CDKAL 1影响糖原储存，并且在HepG 2细胞中使用CDK 5（CDKAL 1的假定靶点）的突变形式的分子实验表明，这是由CDK 5和PKA对MEK的协调调节介导的，MEK最终调节核糖体蛋白S6的磷酸化，这是胰岛素信号传导途径中的关键步骤。
Genome-wide association (GWA) studies have described a large number of new candidate genes that contribute to of Type 2 Diabetes (T2D). In some cases, small clusters of genes are implicated, rather than a single gene, and in all cases, the genetic contribution is not defined through the effects on a specific organ, such as the pancreas or liver. There is a significant need to develop and use human cell-based models to examine the effects these genes may have on glucose regulation. We describe the development of a primary human hepatocyte model that adjusts glucose disposition according to hormonal signals. This model was used to determine whether candidate genes identified in GWA studies regulate hepatic glucose disposition through siRNAs corresponding to the list of identified genes. We find that several genes affect the storage of glucose as glycogen (glycolytic response) and/or affect the utilization of pyruvate, the critical step in gluconeogenesis. Of the genes that affect both of these processes, CAMK1D, TSPAN8 and KIF11 affect the localization of a mediator of both gluconeogenesis and glycolysis regulation, CRTC2, to the nucleus in response to glucagon. In addition, the gene CDKAL1 was observed to affect glycogen storage, and molecular experiments using mutant forms of CDK5, a putative target of CDKAL1, in HepG2 cells show that this is mediated by coordinate regulation of CDK5 and PKA on MEK, which ultimately regulates the phosphorylation of ribosomal protein S6, a critical step in the insulin signaling pathway.
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影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
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5.3
作者：
Arif, Abul;Chatterjee, Piyali;Fox, Paul L.
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Fox, Paul L.
影响因子：
64.8
作者：
Calfon, M;Zeng, HQ;Ron, D
通讯作者：
Ron, D
影响因子：
7.7
作者：
Gregor MF;Yang L;Fabbrini E;Mohammed BS;Eagon JC;Hotamisligil GS;Klein S
通讯作者：
Klein S
影响因子：
4.8
作者：
Hall, RK;Yamasaki, T;Granner, DK
通讯作者：
Granner, DK