Role of SHP-1 Deregulation in HTLV-1 Leukemogenesis

SHP-1 失调在 HTLV-1 白血病发生中的作用

• 批准号: 7229563
• 负责人: Wayne A. Marasco
• 金额: $ 25.54万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7229563
• 关键词: Acetylation; Adult; Apoptosis; Binding; Binding Proteins; Binding Sites; Biochemical; Biological; Biological Assay; CD4 Positive T Lymphocytes; Cell Cycle Progression; Cell Line; Cell Proliferation; Cells; Chromatin; Chromatin Remodeling Factor; Code; Complex; Coupled; Cytokine Signaling; Cytosine; DNA; DNA Methyltransferase; DNA Modification Methylases; Data; Dependence; Development; Disease; Distal; Docking; Down-Regulation; Event; Growth; HDAC1 gene; Histone Code; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone H3; Human; Human T-lymphotropic virus 1; Interleukin 2 Receptor; Interleukin-2; Lead; Leukemic Cell; Maps; Messenger RNA; Methylation; Molecular; Molecular Profiling; Oncogenic; PTPN6 gene; Pathway interactions; Patients; Phosphoric Monoester Hydrolases; Phosphotransferases; Play; Post-Translational Protein Processing; Process; Promoter Regions; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Receptor Signaling; Recruitment Activity; Role; Signal Pathway; Signal Transduction; Site; T-Cell Leukemia; T-Lymphocyte; Taxes; Trans-Activators; Transcription Coactivator; Transcription Repressor/Corepressor; Transferase; Transformed Cell Line; Tyrosine; Virus; Week; adult leukemia; base; cell transformation; chromatin immunoprecipitation; gene replacement; histone methyltransferase; insight; leukemogenesis; promoter; protein expression; receptor expression; tax Gene Products; transcription factor

DESCRIPTION (provided by applicant): Human T-cell lymphotropic virus I (HTLV-I) is the etiological agent for Adult T-cell Leukemia (ATL). Past studies have provided evidence that deregulation of IL-2 receptor signaling may play an important role in the events that lead to immortalization and oncogenic transformation of CD4+ T-cells by HTLV-1. Indeed, constitutive activation of the IL-2R complex including the Jak/STAT pathway is found in HTLV-1 transformed cells and freshly isolated leukemic cells from ATL patients. We and others have recently demonstrated that Shp-1, a constitutively expressed cytosolic protein tyrosine phosphatase that functions as an early negative regulator of IL-2R signaling is selectively and markedly downregulated in HTLV-1 transformed cells. Shp-1 is normally recruited to phosphorylated docking sites on IL-2R where it can then dephosphorylate activated protein tyrosine (Jak) kinases that are part of the signaling complex and we hypothesize that SHP-1 downregulation is of central importance to HTLV-1 leukemogenesis. In this proposal we provide new data which demonstrates by ChIP assay analysis that both the histone deacetylase HDAC1, the histone H3 methyl transferase SUV39H1 and HTLV-1 Tax are bound to the Shp-1 promoter in the HTLV-1 transformed cells. Thus, the molecular basis of Shp-1 promoter silencing appears to be the assembly of a repressor complex involving HDAC1, SUV39H1 and Tax and other unknown proteins at the Shp-1 promoter. In this proposal we will investigate the molecular basis of Shp-1 promoter silencing in these HTLV-1 transformed cells. Specifically, we will first identify and map the core promoter region and transcription factor binding sites in the Shp-1 promoter and determine if transcription factor binding is altered in HTLV-1 transformed and ATL cell lines. We will also determine by ChIP assay if there is binding of transcriptional repressors, methyl-cytosine binding proteins or various chromatin-remodeling complexes to the Shp-1 promoter in the HTLV-1 transformed cell lines. We will also determine if the Shp-1 core promoter region has been methylated. We will also identify the "histone code" of the Shp-1 promoter in fresh CD4+ T-cells and non-HTLV-1 transformed cells and compare this code to HTLV-1 transformed and ATL cell lines. By comparing these codes to the Shp-1 mRNA and protein expression profiles in these cells we will determine if the precise histone codes for Shp-1 expression and silencing can be identified. We will correlate these findings to the ChIP assay analyses of chromatin modifying proteins to determine if the biochemical mechanisms involved in Shp-1 promoter silencing can be elucidated. We will also determine the molecular interactions that lead to HTLV-1 Tax binding to the Shp-1 promoter and will evaluate post-translational modifications to Tax (e.g. acetylation, methylation) that may influence preferential binding to transcription factors, coactivators or corepressors. Finally, we will examine the effects of inhibitors of histone deacetylase and DNA methyltransferase on activation of the Shp-1 promoter in HTLV-1 transformed cell lines. We will also investigate the biological consequences of this inhibition on Shp-1 expression and constitutive activation of the Jak/STAT pathway. These studies will be coupled with Shp-1 gene replacement studies to evaluate the longer term consequences of Shp-1 re-expression on cell proliferation, viability, cell cycle progression and apoptosis. These studies should provide new insight into the biochemical mechanisms of HTLV-1 leukemogenesis and aid in the development of new treatments for this fatal disease.

描述（由申请方提供）：人T细胞嗜淋巴细胞病毒I（HTLV-I）是成人T细胞白血病（ATL）的病原体。过去的研究已经提供了证据表明，IL-2受体信号转导的失调可能在HTLV-1导致CD 4 + T细胞永生化和致癌转化的事件中起重要作用。实际上，在HTLV-1转化的细胞和来自ATL患者的新鲜分离的白血病细胞中发现了包括Jak/STAT途径的IL-2 R复合物的组成性活化。我们和其他人最近已经证明，Shp-1，组成型表达的胞质蛋白酪氨酸磷酸酶，作为一个早期的IL-2 R信号负调节剂的功能是选择性和显着下调HTLV-1转化细胞。Shp-1通常被募集到IL-2 R上的磷酸化对接位点，在那里它可以使作为信号复合物一部分的活化蛋白酪氨酸（Jak）激酶去磷酸化，我们假设SHP-1下调对HTLV-1白血病发生至关重要。在该提案中，我们提供了新的数据，其通过ChIP测定分析证明组蛋白脱乙酰酶HDAC 1、组蛋白H3甲基转移酶SUV 39 H1和HTLV-1 Tax都与HTLV-1转化细胞中的Shp-1启动子结合。因此，Shp-1启动子沉默的分子基础似乎是在Shp-1启动子处组装涉及HDAC 1、SUV 39 H1和Tax以及其他未知蛋白质的阻遏物复合物。在这个提议中，我们将研究这些HTLV-1转化细胞中Shp-1启动子沉默的分子基础。具体来说，我们将首先确定和映射的核心启动子区域和转录因子结合位点的Shp-1启动子，并确定是否转录因子结合改变HTLV-1转化和ATL细胞系。我们还将通过ChIP测定确定HTLV-1转化细胞系中是否存在转录抑制因子、甲基胞嘧啶结合蛋白或各种染色质重塑复合物与Shp-1启动子的结合。我们还将确定Shp-1核心启动子区域是否已被甲基化。我们还将鉴定新鲜的CD 4 + T细胞和非HTLV-1转化细胞中Shp-1启动子的“组蛋白密码”，并将该密码与HTLV-1转化细胞和ATL细胞系进行比较。通过比较这些代码在这些细胞中的Shp-1 mRNA和蛋白质表达谱，我们将确定是否可以确定精确的组蛋白编码的Shp-1表达和沉默。我们将这些发现与染色质修饰蛋白的ChIP分析相关联，以确定是否可以阐明Shp-1启动子沉默所涉及的生化机制。我们还将确定导致HTLV-1 Tax与Shp-1启动子结合的分子相互作用，并将评估Tax的翻译后修饰（例如乙酰化、甲基化），这些修饰可能影响与转录因子、辅激活因子或辅阻遏因子的优先结合。最后，我们将研究组蛋白去乙酰化酶和DNA甲基转移酶抑制剂对HTLV-1转化细胞系中Shp-1启动子激活的影响。我们还将研究这种抑制对Shp-1表达和Jak/STAT通路组成性激活的生物学后果。这些研究将与Shp-1基因替代研究相结合，以评估Shp-1重新表达对细胞增殖、活力、细胞周期进展和凋亡的长期影响。这些研究将为HTLV-1白血病发生的生化机制提供新的见解，并有助于开发这种致命疾病的新治疗方法。