Nuclear Translocation of I-kappaB-alpha as a Therapeutic Target

I-κB-α 的核转位作为治疗靶点

• 批准号: 7184005
• 负责人: Ivana Vancurova
• 金额: $ 24.53万
• 依托单位: ST. JOHN'S UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-01-09 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7184005
• 关键词: Affinity Chromatography; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Apoptotic; Binding; Biochemical; Cell Nucleus; Cells; Class; Confocal Microscopy; Data; Development; Disease; Drug Delivery Systems; Event; Fractionation; Gene Expression; Genes; Genetic Transcription; Goals; HL-60 Cells; HL60; Hela Cells; Human; Immunoprecipitation; Inflammatory; Laboratories; Leukocytes; MG115; MG132; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Molecular; Nuclear; Nuclear Matrix; Nuclear Protein; Nuclear Proteins; Nuclear Translocation; Post-Translational Protein Processing; Proteasome Inhibition; Proteasome Inhibitor; Proteins; RNA; Rate; Rest; Role; Testing; Transfection; base; cell type; chromatin immunoprecipitation; human disease; inhibitor/antagonist; leukemia; monocyte; multicatalytic endopeptidase complex; neutrophil; novel; promoter; protein expression; therapeutic target; transcription factor

DESCRIPTION (provided by applicant): NFkB is a critical transcription factor regulating expression of pro-inflammatory and anti- apoptotic genes. Previous studies from our laboratory demonstrated that as opposed to other cell types, resting human neutrophils contain predominant amount of NFkB inhibitor, IkBa, in the nucleus, and this increased nuclear accumulation of IkBa results in the inhibition of NFkB activity and increased rate of neutrophil apoptosis. Our recent data have shown that proteasome inhibition induces translocation of IkBa to the nucleus in leukemia HL-60 and U- 937 cells, and in cancer HeLa cells; however, the mechanisms are currently unknown. The central hypothesis of this proposal is that induction of nuclear accumulation of IkBa inhibits NFkB activity and expression of NFkB-regulated anti-apoptotic and pro-inflammatory genes, and could thus provide a basis for novel anti-cancer and anti-inflammatory therapies aimed at the inhibition of NFkB activity by the nuclear IkBa. The specific aims focus on analyzing the mechanisms by which the nuclear IkBa inhibits NFkB activity and expression of NFkB-regulated genes in leukemia HL-60 and U-937 cells. In Aim 1, we will test the hypothesis that the proteasome inhibition-induced nuclear translocation of IkBa results in the inhibition of NFkB activity and decreased expression of NFkB-regulated anti-apoptotic genes, by using transfection of HL-60 and U-937 cells with inhibitory IkBa RNA. In Aim 2, we will use chromatin immunoprecipitation to test the hypothesis that the nuclear IkBa inhibits NFkB activity by associating with the promoters of NFkB-regulated genes. In Aim 3, we will investigate the mechanisms that regulate the proteasome inhibition-induced nuclear translocation of IkBa. Identification of the key molecular events that control NFkB activity by the nuclear IkBa will broaden our understanding of the mechanisms regulating NFkB activity, and might provide a new class of drug targets to regulate the NFkB driven pro-inflammatory and anti-apoptotic gene expression. The focus of this proposal is to identify the mechanisms by which the nuclear translocation of IkBa regulates activity of the transcription factor NFkB. Since NFkB activity is increased in many human diseases including inflammatory disorders, cancer, and leukemia, identification of the mechanisms by which the nuclear IkBa inhibits NFkB activity will contribute to the development of novel anti-cancer and anti-inflammatory therapies.

描述（由申请人提供）：NFKB是调节促炎和抗凋亡基因表达的关键转录因子。我们实验室的先前研究表明，与其他细胞类型相反，静止的人类嗜中性粒细胞含有主要量的NFKB抑制剂IKBA，IKBA，以及IKBA的核积累增加导致NFKB活性的抑制和中性粒细胞增生的速率增加。我们最近的数据表明，蛋白酶体抑制会诱导IKBA易位到白血病HL-60和U-937细胞以及癌症HeLa细胞中的核。但是，这些机制目前尚不清楚。该提议的中心假设是，IKBA核积累的诱导抑制了NFKB的活性并表达了NFKB调节的抗凋亡和促炎基因，因此可以为新型抗癌和抗炎疗法提供基础，旨在抑制NFKB活性受核IKBA抑制NFKB活性。该特定目的是分析核IKBA抑制NFKB活性和NFKB调节基因在白血病HL-60和U-937细胞中的表达的机制。在AIM 1中，我们将检验以下假设：蛋白酶体抑制作用诱导的IKBA核易位会导致NFKB活性的抑制作用，并使用HL-60和U-937细胞抑制IKBA RNA，从而抑制NFKB活性并降低NFKB调节的抗凋亡基因的表达。在AIM 2中，我们将使用染色质免疫沉淀来检验以下假设：核IKBA通过与NFKB调节的基因的启动子相关联抑制NFKB活性。在AIM 3中，我们将研究调节蛋白酶体抑制引起的IKBA核易位的机制。通过核IKBA控制NFKB活性的关键分子事件将扩大我们对调节NFKB活性的机制的理解，并可能提供新的药物靶标，以调节NFKB驱动的促炎和抗凋亡基因表达。该提案的重点是确定IKBA核易位调节转录因子NFKB活性的机制。由于包括炎症性疾病，癌症和白血病在内的许多人类疾病中的NFKB活性增加，因此识别核IKBA抑制NFKB活性的机制将有助于新型抗癌和抗炎疗法的发展。

项目成果

Bortezomib induces nuclear translocation of IκBα resulting in gene-specific suppression of NF-κB--dependent transcription and induction of apoptosis in CTCL.

• DOI: 10.1158/1541-7786.mcr-10-0368
• 发表时间: 2011-02
• 期刊: Molecular cancer research : MCR
• 作者: Juvekar A;Manna S;Ramaswami S;Chang TP;Vu HY;Ghosh CC;Celiker MY;Vancurova I
• 通讯作者: Vancurova I