Rare coding variants of the adenosine A3 receptor are increased in autism: on the trail of the serotonin transporter regulome.
Rare coding variants of the adenosine A3 receptor are increased in autism: on the trail of the serotonin transporter regulome.
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DOI：
10.1186/2040-2392-4-28
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发表时间：
2013-08-16
期刊：
影响因子：
6.2
通讯作者：
Sutcliffe JS
中科院分区：
文献类型：
--
作者：
Campbell NG;Zhu CB;Lindler KM;Yaspan BL;Kistner-Griffin E;NIH ARRA Consortium;Hewlett WA;Tate CG;Blakely RD;Sutcliffe JS
罕见的遗传变异是自闭症谱系障碍(ASD)的一类重要危险因素，并可牵涉到生物网络进行研究。5-羟色胺(5-HT)信号的改变与ASD有关，我们和其他人在5-HT转运体(SERT)基因中发现了多种罕见的ASD相关变异，导致5-HT重摄取增加和调节紊乱。我们假设编码SERT调节器的基因座含有影响SERT功能和/或调节的变异，因此可能导致ASD风险。腺苷A3受体(A3AR)通过蛋白激酶G(PKG)等信号通路调节SERT，导致SERT表面表达增强和催化活性增强。为了验证我们的假设，我们询问了A3AR基因的罕见变异(ADORA3)在ASD病例中是否比对照组增加。对病例对照样本进行了发现测序，并随后对比较外显子组序列数据进行了分析。我们评估了发现样本中的两个变体对A3AR信号和SERT活性的功能影响。测序结果显示，病例组与对照组相比，罕见的编码变异增加(P=0.013)。虽然比较外显子组序列数据没有显示显著的富集性(P=0.071)，但联合分析加强了相关性的证据(P=0.0025)。在ASD病例中发现的两个变异体(Leu90Val和Val171Ile)位于配体结合口袋内或附近，且Leu90Val在病例中单独富含(P=0.040)。对表达Val90-A3AR的细胞的体外分析显示，与野生型受体相比，基础cGMP水平升高。此外，与野生型受体相比，一种特定的A3AR激动剂在研究的整个过程中增加了Val90-A3AR细胞中cGMP的水平。在Val90-A3AR/SERT共转染中，激动剂刺激提高了野生型受体上的SERT活性，并延迟了5-羟色胺摄取活性的恢复。相反，Ile171-A3AR不能支持激动剂刺激SERT。虽然Val90和Ile171在这些ASD病例中的出现数量较多，但家系分离分析显示不完全外显，与其他罕见的ASD风险等位基因一致。我们的结果证实了SERT调节网络含有影响ASD中SERT活性和调节的罕见功能变异的假设，并鼓励进一步研究该网络中可能影响ASD风险的其他变异。
Rare genetic variation is an important class of autism spectrum disorder (ASD) risk factors and can implicate biological networks for investigation. Altered serotonin (5-HT) signaling has been implicated in ASD, and we and others have discovered multiple, rare, ASD-associated variants in the 5-HT transporter (SERT) gene leading to elevated 5-HT re-uptake and perturbed regulation. We hypothesized that loci encoding SERT regulators harbor variants that impact SERT function and/or regulation and therefore could contribute to ASD risk. The adenosine A3 receptor (A3AR) regulates SERT via protein kinase G (PKG) and other signaling pathways leading to enhanced SERT surface expression and catalytic activity. To test our hypothesis, we asked whether rare variants in the A3AR gene (ADORA3) were increased in ASD cases vs. controls. Discovery sequencing in a case-control sample and subsequent analysis of comparison exome sequence data were conducted. We evaluated the functional impact of two variants from the discovery sample on A3AR signaling and SERT activity. Sequencing discovery showed an increase of rare coding variants in cases vs. controls (P=0.013). While comparison exome sequence data did not show a significant enrichment (P=0.071), combined analysis strengthened evidence for association (P=0.0025). Two variants discovered in ASD cases (Leu90Val and Val171Ile) lie in or near the ligand-binding pocket, and Leu90Val was enriched individually in cases (P=0.040). In vitro analysis of cells expressing Val90-A3AR revealed elevated basal cGMP levels compared with the wildtype receptor. Additionally, a specific A3AR agonist increased cGMP levels across the full time course studied in Val90-A3AR cells, compared to wildtype receptor. In Val90-A3AR/SERT co-transfections, agonist stimulation elevated SERT activity over the wildtype receptor with delayed 5-HT uptake activity recovery. In contrast, Ile171-A3AR was unable to support agonist stimulation of SERT. Although both Val90 and Ile171 were present in greater numbers in these ASD cases, segregation analysis in families showed incomplete penetrance, consistent with other rare ASD risk alleles. Our results validate the hypothesis that the SERT regulatory network harbors rare, functional variants that impact SERT activity and regulation in ASD, and encourages further investigation of this network for other variation that may impact ASD risk.
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影响因子：
16.2
作者：
Iossifov I;Ronemus M;Levy D;Wang Z;Hakker I;Rosenbaum J;Yamrom B;Lee YH;Narzisi G;Leotta A;Kendall J;Grabowska E;Ma B;Marks S;Rodgers L;Stepansky A;Troge J;Andrews P;Bekritsky M;Pradhan K;Ghiban E;Kramer M;Parla J;Demeter R;Fulton LL;Fulton RS;Magrini VJ;Ye K;Darnell JC;Darnell RB;Mardis ER;Wilson RK;Schatz MC;McCombie WR;Wigler M
通讯作者：
Wigler M
影响因子：
5.2
作者：
Horvath, S;Xu, X;Laird, NM
通讯作者：
Laird, NM
影响因子：
64.8
作者：
Cook, Edwin H., Jr.;Scherer, Stephen W.
通讯作者：
Scherer, Stephen W.
影响因子：
14.9
作者：
Bromberg Y;Rost B
通讯作者：
Rost B
影响因子：
9.8
作者：
Li, Bingshan;Leal, Suzanne M.
通讯作者：
Leal, Suzanne M.