The Role of Zinc Finger Genes in Leukemogenesis

锌指基因在白血病发生中的作用

• 批准号: 8490414
• 负责人: Sinisa Dovat
• 金额: $ 35.97万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8490414
• 关键词: Adolescent; Adult Lymphoma; Amino Acids; Binding; Biochemical; Biological; Biological Process; CD8B1 gene; Cell Cycle Progression; Cell Cycle Regulation; Cell Proliferation; Cell physiology; Cells; Childhood; Childhood Acute Lymphocytic Leukemia; Chronic Lymphocytic Leukemia; DNA Binding; Data; Development; Down-Regulation; Exhibits; Functional disorder; G1 Phase; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Hematopoietic; Human; Ikaros protein; Immune; In Vitro; Infant; Interleukin-2; Knockout Mice; Lymphoma; Malignant - descriptor; Malignant Neoplasms; Maps; Molecular; Mus; Nucleic Acid Regulatory Sequences; Patients; Peripheral; Phosphorylation; Phosphorylation Site; Phosphotransferases; Process; Protein Dephosphorylation; Proteins; RNA Interference; RNA Splicing; Regulation; Regulatory Element; Research; Retroviral Vector; Role; S Phase; Signal Transduction; Signal Transduction Pathway; Site; Stem cells; System; T cell differentiation; T-Cell Proliferation; T-Lymphocyte; Testing; Transcriptional Regulation; Transplantation; Tumor Suppressor Proteins; Up-Regulation; Zinc Fingers; adult leukemia; bone marrow hyperplasia; chromatin remodeling; effective therapy; in vivo; insight; leukemia; leukemia/lymphoma; leukemogenesis; mouse model; mutant; novel; overexpression; promoter; protein protein interaction; reconstitution; research study; response; thymocyte

PROJECT SUMMARY The objective of the proposed study is to understand the malignant transformation of hematopoietic cells by identifying the biological functions of Ikaros proteins. Ikaros is essential for hematopoietic development and acts as a tumor suppressor. The Ikaros gene is alternately spliced to generate multiple zinc finger proteins involved in gene regulation and chromatin remodeling. Our preliminary data show that: 1) Ikaros is phosphorylated at multiple evolutionarily conserved sites by CK2 and other kinases during G1 and S phase of the cell cycle 2) Phosphorylation of Ikaros at specific amino acids regulates its DNA-binding ability and subcellular localization. 3) Ikaros binds to the Bcl-xL gene promoter in vivo and disregulation of Ikaros activity is associated with upregulation of Bcl-xL gene expression. The specific aims of our proposals are: Specific Aim #1: To identify the specific phosphorylation sites responsible for Ikaros function in the regulation of transcription, cellular proliferation and differentiation. We hypothesize that phosphorylation of Ikaros interferes with its function in transcriptional regulation and chromatin remodeling and influences cellular proliferation. We will define phosphorylation sites that are critical for Ikaros function in DNA-binding, subcellular localization, and protein-protein interaction. The role of Ikaros phosphorylation in controlling cell cycle progression will be studied using a murine leukemia cellular system derived from Ikaros deficient mice. These cells will be transduced with wild type or Ikaros phosphomimetic mutants to define sites that are critical for Ikaros' function in cell cycle control. The role of Ikaros' phosphorylation in regulating T cell differentiation and T cell proliferation will be studied in vivo. Murine stem cells from mice with targeted disruption of Ikaros will be infected with retroviral vectors containing wild type Ikaros or phosphomimetic Ikaros mutants and transplanted into sublethally irradiated Ikaros knockout mice. The ability of phosphomimetic Ikaros mutants to restore normal T cell differentiation will be compared to that of wild type Ikaros. Specific aim #2: To dissect the mechanism by which Ikaros regulates Bcl-xL expression. Previous studies suggest that decreased Ikaros activity leads to overexpression of the Bcl-xL gene. We hypothesize that Ikaros exerts its tumor suppressor activity by negatively regulating Bcl-xL expression. To test this hypothesis we will determine whether increased Ikaros expression downregulates Bcl-xL transcription in human lymphoma cells and we will map the regions of the Bcl-xL upstream regulatory element (URE) that are critical for Ikaros-modulated control of Bcl-xL expression. These studies will provide the first detailed functional analysis of the signal transduction pathways that control the tumor suppressor function of Ikaros. Our research will provide new and important information on the mechanisms controlling the proliferation of hematopoietic cells and will yield insights into the pathophysiology and treatment of leukemia.

项目摘要 拟议的研究的目的是了解造血细胞的恶性转化 通过识别Ikaros蛋白的生物学功能。 Ikaros对于造血发展至关重要 充当肿瘤抑制剂。 Ikaros基因交替剪接以产生多个锌指蛋白 参与基因调节和染色质重塑。 我们的初步数据表明：1）Ikaros通过多个进化保守的地点磷酸化 在细胞周期的G1和S期间的CK2和其他激酶2）特定氨基处的Ikaros磷酸化 酸调节其DNA结合能力和亚细胞定位。 3）Ikaros与Bcl-XL基因启动子结合 体内和ikaros活性的脱离与Bcl-XL基因表达的上调有关。 我们建议的具体目的是：特定目的＃1：确定特定的磷酸化位点 负责IKAROS功能在转录，细胞增殖和分化的调节中。我们 假设Ikaros的磷酸化会干扰其在转录调控和 染色质重塑并影响细胞增殖。我们将定义关键的磷酸化位点 对于Ikaros的功能，在DNA结合，亚细胞定位和蛋白质 - 蛋白质相互作用中。伊卡罗斯的角色 将使用鼠白血病细胞系统研究控制细胞周期进程中的磷酸化 源自Ikaros缺乏的小鼠。这些细胞将用野生型或ikaros磷酸化转导 突变体定义对Ikaros在细胞周期控制中功能至关重要的位点。伊卡罗斯的角色 调节T细胞分化和T细胞增殖的磷酸化将在体内研究。鼠茎 来自Ikaros靶向破坏的小鼠的细胞将被含有野生型的逆转录病毒载体感染 Ikaros或磷酸化Ikaros突变体，并移植到共辐射的Ikaros基因敲除小鼠中。 将磷酸化Ikaros突变体恢复正常T细胞分化的能力将与 野生型Ikaros。特定目的＃2：剖析Ikaros调节Bcl-XL表达的机制。 先前的研究表明，Ikaros活性降低会导致BCL-XL基因的过表达。我们 假设Ikaros通过负调节BCl-XL表达来发挥其肿瘤抑制活性。测试 该假设我们将确定增加Ikaros表达是否下调BCl-XL转录 人淋巴瘤细胞，我们将绘制BCL-XL上游调节元件（URE）的区域 对于Ikaros调节BCl-XL表达的控制至关重要。 这些研究将对信号转导途径进行第一个详细的功能分析 控制Ikaros的肿瘤抑制功能。我们的研究将提供有关有关的新的重要信息 控制造血细胞增殖的机制，并将对病理生理学产生见解 和白血病的治疗。

项目成果

Induction of senescence by adenosine suppressing the growth of lung cancer cells.

• DOI: 10.1016/j.bbrc.2013.09.030
• 发表时间: 2013-10
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Dongqin Yang;Junyao Song;Lijun Wu;Yunfang Ma;Chunhua Song;S. Dovat;T. Nishizaki;Jie Liu

Pathogenesis and regulation of cellular proliferation in acute lymphoblastic leukemia - the role of Ikaros.

急性淋巴细胞白血病的发病机制和细胞增殖调节 - Ikaros 的作用。

• 发表时间: 2014
• 期刊: Journal of B.U.ON. : official journal of the Balkan Union of Oncology
• 作者: Wang,Haijun;Ouyang,Hongsheng;Lai,Liangxue;Petrovic-Dovat,Lidija;Stankov,Karmen;Bogdanovic,Gordana;Dovat,Sinisa
• 通讯作者: Dovat,Sinisa

Ikaros, CK2 kinase, and the road to leukemia.

• DOI: 10.1007/s11010-011-0964-5
• 发表时间: 2011-10
• 期刊: MOLECULAR AND CELLULAR BIOCHEMISTRY
• 影响因子: 4.3
• 作者: Dovat, Sinisa;Song, Chunhua;Payne, Kimberly J.;Li, Zhanjun
• 通讯作者: Li, Zhanjun

AMID mediates adenosine-induced caspase-independent HuH-7 cell apoptosis.

AMID 介导腺苷诱导的不依赖 caspase 的 HuH-7 细胞凋亡。

• DOI: 10.1159/000325203
• 发表时间: 2011
• 期刊: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology
• 作者: Yang,Dongqin;Yaguchi,Takahiro;Nagata,Tetsu;Gotoh,Akinobu;Dovat,Sinisa;Song,Chunhua;Nishizaki,Tomoyuki
• 通讯作者: Nishizaki,Tomoyuki

Regulation of Ikaros function by casein kinase 2 and protein phosphatase 1.

• DOI: 10.4331/wjbc.v2.i6.126
• 发表时间: 2011-06-26
• 期刊: World journal of biological chemistry
• 作者: Song, Chunhua;Li, Zhanjun;Dovat, Sinisa
• 通讯作者: Dovat, Sinisa