The Role of Zinc Finger Genes in Leukemogenesis

锌指基因在白血病发生中的作用

基本信息

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
项目类别：
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

项目摘要

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.

项目概要拟议研究的目的是了解造血细胞的恶性转化通过鉴定 Ikaros 蛋白的生物学功能。 Ikaros 对于造血发育至关重要作为肿瘤抑制剂。 Ikaros基因交替剪接生成多种锌指蛋白参与基因调控和染色质重塑。我们的初步数据表明：1) Ikaros 在多个进化保守位点被磷酸化细胞周期 G1 和 S 期的 CK2 和其他激酶 2) Ikaros 在特定氨基处的磷酸化酸调节其 DNA 结合能力和亚细胞定位。 3) Ikaros结合Bcl-xL基因启动子 Ikaros 活性的失调与 Bcl-xL 基因表达的上调有关。我们提案的具体目标是：具体目标#1：确定特定的磷酸化位点负责 Ikaros 调节转录、细胞增殖和分化的功能。我们假设 Ikaros 的磷酸化会干扰其转录调控功能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 Ikaros 在 DNA 结合、亚细胞定位和蛋白质-蛋白质相互作用中发挥作用。伊卡洛斯角色将使用鼠白血病细胞系统研究控制细胞周期进程的磷酸化源自 Ikaros 缺陷小鼠。这些细胞将用野生型或 Ikaros 磷模拟物转导突变体来定义对 Ikaros 在细胞周期控制中的功能至关重要的位点。伊卡洛斯的角色将在体内研究调节 T 细胞分化和 T 细胞增殖的磷酸化。鼠干来自靶向破坏 Ikaros 的小鼠的细胞将被含有野生型的逆转录病毒载体感染 Ikaros 或拟磷 Ikaros 突变体并移植到亚致死照射的 Ikaros 基因敲除小鼠中。拟磷化 Ikaros 突变体恢复正常 T 细胞分化的能力将与野生型伊卡洛斯。具体目标#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 基因过度表达。我们假设 Ikaros 通过负向调节 Bcl-xL 表达来发挥其肿瘤抑制活性。测试根据这一假设，我们将确定 Ikaros 表达增加是否会下调 Bcl-xL 转录人类淋巴瘤细胞，我们将绘制 Bcl-xL 上游调节元件 (URE) 的区域，这些区域是对于 Ikaros 调节的 Bcl-xL 表达控制至关重要。这些研究将对信号转导途径进行首次详细的功能分析，控制Ikaros的抑癌功能。我们的研究将提供有关以下方面的新的重要信息控制造血细胞增殖的机制并将深入了解病理生理学和治疗白血病。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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：
发表时间：
2011
期刊：
影响因子：
0
作者：
Yang, Dongqin;Yaguchi, Takahiro;Nagata, Tetsu;Gotoh, Akinobu;Dovat, Sinisa;Song, Chunhua;Nishizaki, Tomoyuki
通讯作者：
Nishizaki, Tomoyuki

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

腺苷诱导衰老抑制肺癌细胞的生长。

DOI：
10.1016/j.bbrc.2013.09.030
发表时间：
2013-10-11
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
通讯作者：
Jie Liu

Ikaros isoforms: The saga continues.

Ikaros 同种型：传奇仍在继续。

DOI：
发表时间：
2011-06-26
期刊：
影响因子：
0
作者：
Li, Zhanjun;Perez;Savic, Aleksandar;Song, Chunhua;Dovat, Sinisa
通讯作者：
Dovat, Sinisa

Transcriptional Regulation of JARID1B/KDM5B Histone Demethylase by Ikaros, Histone Deacetylase 1 (HDAC1), and Casein Kinase 2 (CK2) in B-cell Acute Lymphoblastic Leukemia.

B 细胞急性淋巴细胞白血病中 Ikaros、组蛋白脱乙酰酶 1 (HDAC1) 和酪蛋白激酶 2 (CK2) 对 JARID1B/KDM5B 组蛋白脱甲基酶的转录调节。

DOI：
发表时间：
2016-02-19
期刊：
影响因子：
0
作者：
Wang, Haijun;Song, Chunhua;Ding, Yali;Pan, Xiaokang;Ge, Zheng;Tan, Bi;Gowda, Chandrika;Sachdev, Mansi;Muthusami, Sunil;Ouyang, Hongsheng;Lai, Liangxue;Francis, Olivia L;Morris, Christopher L;Abdel;Dorsam, Glenn;Xiang, Meixi
通讯作者：
Xiang, Meixi

Genetic targets in pediatric acute lymphoblastic leukemia.

小儿急性淋巴细胞白血病的遗传靶点。