2011-01-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659203摘要：在許多發炎相關疾病及癌症中可以發現到IKK/NF-kB活性增加，而轉錄共同活化因子CBP，其具有histone acetyltransferase (HAT)活性，被認為與胚胎發育有關，且是一種腫瘤抑制因子，CBP 基因突變會造成發育缺陷與Rubinstein-Taybi Syndrome(RTS)，此與CBP 之HAT活性受到抑制有關。我們最近的研究證明，IKKa磷酸化CBPSer1382/1386（在小鼠中為Ser1364/1368）而增加其HAT 與轉錄活性，然而，其生理上所代表的意義尚未被釐清，為了進一步研究，我們將培育conventional 與conditionalCBP-SSAA knockin mice，其Ser1364/1368 (SS)被置換成無法被磷酸化之alanines(AA)，我們的初步結果發現CBP+/AA mice表現出顏面異常及生長遲緩的現象，此與RTS的病徵類似，代表CBP 磷酸化對於生長發育非常重要。此外，因為RTS 病人罹患癌症的風險較高，我們將進一步以conditional CBP-SSAA knockin mice 之colitis-associatedcancer模式，來研究wild-type與CBP-SSAA knockin mice產生腫瘤的比例。此兩年期計畫將研究CBP 磷酸化在發育及發炎相關癌化中所扮演的角色，其目標為：第一年：1. 培育conventional與conditional CBP-SSAA knockin mice，並分析其基因型與表現型。我們預期heterozygous CBP-SSAA knockin mice 會表現出RTS 的病徵，而homozygous CBP-SSAA knockin mice會死於胚胎發育階段。2. 研究在in vivo情況下，IKKa是否磷酸化CBP之Ser1364/1368。我們預期CBP 磷酸化增加其HAT與轉錄活性，且是發育過程所需。第二年：3. 研究CBP Ser1364/1368磷酸化缺失是否會表現出Rubinstein-Taybi Sydrome (RTS)之臨床病徵，以及HDAC抑制劑是否有保護效果。我們將進一步以行為測驗來分析RTS 之臨床病徵，我們預期CBP-SSAA mice將顯露出許多行為上的缺陷，給予HDAC 抑制劑可以回復此現象。4. 研究CBP Ser1364/1368磷酸化在colon-specific CBP-SSAA knockin小鼠之發炎相關癌症模式之效應。我們將培育colon-specific CBP-SSAA knockin mice 來研究其在colitis-associatedcancer 所扮演之角色，我們預期CBP-SSAA knockin mice 之腫瘤發生率將低於wild-type mice。<br> Abstract: Elevated levels of IKK/NF-kB are frequently detected in many inflammatory diseasesand cancers. The transcriptional co-activator CBP possessing intrinsic histoneacetyltransferase (HAT) activity has been implicated in embryonic development and servesas a tumor suppressor. Aberrant mutations of CBP gene result in developmental disorder andRubinstein-Taybi syndrome (RTS), which is correlated with defect in HAT activity. We haverecently demonstrated that IKKa phosphorylates CBP at Ser1382/1386 (Ser1364/1368 inmouse), which enhances its HAT and transcriptional activities. However, the physiologicalsignificance of CBP phosphorylation at these sites is unknown. To investigate this,conventional and conditional CBP-SSAA knockin mice with the replacement ofSer1364/1368 (SS) to alanines (AA) will be generated. Our preliminary data showed that theconventional heterozygous F1 mice (CBP+/AA) displayed facial abnormalities and growthretardation, which is similar to the phenotypes of RTS, suggesting that CBP phosphorylationmight be important for the development. Since RTS patients have an increased risk for tumor,we will investigate this by a colitis-associated cancer model in conditional CBP-SSAAknockin mice. The exact role of CBP phosphorylation in the development andinflammation-associated tumorigenesis will be examined in this two-year plan. The specificaims are:Year 1:1. To generate conventional and conditional CBP-SSAA knockin mice and examinetheir genotypes and phenotypes.We expect that the heterozygous CBP-SSAA knockin mice will show the features ofRTS and homozygous CBP-SSAA knockin mice will die during embryogenesis.2. To investigate whether IKKa phosphorylates CBP at Ser1364/1368 in vivo.We expect that CBP phosphorylation results in increases of its HAT and transcriptionalactivities, which is required for the development.Year 2:3. To investigate whether deficit of CBP phosphorylation at Ser1364/1368 showsclinical features of Rubinstein-Taybi Syndrome (RTS) and whether HDACinhibitors provide protection.We will further analyze the clinical features of RTS by behavioral tests. We expect thatCBP-SSAA mice will show multiple defects in these tests, which can be reversed byHDAC inhibitors.4. To investigate the effect of CBP phosphorylation at Ser1364/1368 in inflammationassociatedcancer by colon-specific CBP-SSAA knockin mouse model.We will generate colon-specific CBP-SSAA knockin mice to investigate its role incolitis-associated cancer. We expect that the incidence of colitis-associated cancer inCBP-SSAA knockin mice will be lower than wild-type mice.發炎反應輔助轉錄因子CBPT細胞發育inflammationCBPT cell development