Modulation of IFN action via novel regulatory factors

通过新的调节因子调节干扰素的作用

• 批准号: 8197723
• 负责人: DHAN V. KALVAKOLANU
• 金额: $ 31.97万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-16 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8197723
• 关键词: Activating Transcription Factor 2; Antigen Presentation; Antitumor Response; Antiviral Response; Apoptosis; Apoptosis Regulator; Apoptotic; Autophagocytosis; Binding; Biological Assay; Biological Process; Biological Response Modifiers; CCAAT-Enhancer-Binding Protein-beta; CCAAT-Enhancer-Binding Proteins; Cell Cycle; Cell Death; Cell Differentiation process; Cell Line; Cells; Cessation of life; Clinical; Collaborations; Complex; DAP kinase; Data; Defect; Disease; Elements; Endoplasmic Reticulum; Energy Metabolism; Enhancers; Excision; Family; Functional disorder; Funding; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; General Transcription Factors; Genes; Genetic Transcription; Gluconeogenesis; Growth; Hematopoietic; Host Defense; Human; Immune response; Immunity; Interferons; Investigation; Knockout Mice; Knowledge; Laboratories; MAP3K5 gene; Malignant Neoplasms; Mediating; Mediator of activation protein; Mitogen-Activated Protein Kinases; Molecular; Mouse Protein; Mus; Mutagenesis; Neoplasm Metastasis; Neoplasms; Neurodegenerative Disorders; Organelles; Pathway interactions; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiological Processes; Play; Post-Translational Protein Processing; Process; Production; Protein Kinase; Proteins; Proteomics; RNA Interference; Recruitment Activity; Regulation; Regulatory Pathway; Role; STAT1 gene; Shapes; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Stress; TATA Box; Testing; Therapeutic; Time; Transact; Tumor Suppression; Virus Diseases; ZIP kinase; activating transcription factor; base; biological adaptation to stress; cell growth; cell growth regulation; combat; cytokine; design; fighting; in vivo; lipid biosynthesis; macrophage; member; mutant; neoplastic cell; novel; novel therapeutics; pathogen; promoter; public health relevance; response; surveillance network; transcription factor; tumor; tumor growth; tumor progression

DESCRIPTION (provided by applicant): The interferon family of cytokines is critical for promoting several physiologic processes, such as antiviral, antitumor, and immune responses. They are in clinical use for the therapy of a number of cancers, viral diseases and neurodegenerative disorders. By interacting with other cytokines IFNs form a large network of intercellular signaling molecules that control neoplastic cell growth and host defenses against pathogens. Previously, we have identified a novel IFN-regulated element and its cognate transcription factors. One such protein is CCAAT/Enhancer Binding protein-beta (C/EBP-2), a transcription factor known to regulate cell differentiation, energy metabolism, immune response, tumor growth and apoptosis. A gene expression micro- array analysis in our lab identified several IFN regulated genes, whose expression required C/EBP-2. One of them is the death associated protein kinase1 (DAPK1), an important regulator of apoptosis, cell cycle, and metastasis. The expression of dapk1 gene is frequently lost in several human cancers. DAPK1 also regulates autophagy (a novel form of death), which is critical for the removal of damaged organelle, fighting intracellular pathogens, antigen presentation and tumor suppression. Interestingly, the loss of C/EBP-2 gene in mice causes many of these defects. During the last funding period, we have shown a central role for C/EBP-2 in regulating DAPK1. In preliminary studies, we show that the expression of DAPK1 requires other transacting factors. Notably, Activating transcription factor 6 (ATF6), a key regulator of endoplasmic reticulum-dependent stress responses, appears to regulate DAPK1 expression. We propose that a direct interaction between ATF6 and C/EBP-2 leads to DAPK1 expression and growth suppression via autophagy, wherein the MAP Kinase, apoptosis- stimulating kinase 1(ASK1), provides critical signal inputs. In specific aim 1 of this proposal we will investigate how C/EBP-2 and ATF6 collaborate to upregulate DAPK1 expression. In specific aim 2, we will investigate how ASK1 controls DAPK1 expression via ATF6. Enhancer bound transcription factors (TFs) promote transcription using transcriptional co-activators. One of them the Mediator, a molecular bridge comprised of multiple proteins, communicates the transcriptional signals from the TFs to general transcription factor complex at the TATA-box. Studies during the last funding period also identified Med1, a major subunit of Mediator, an IFN-induced binding partner of C/EBP- 2. We present preliminary evidence for the involvement of ZIP-kinase(ZIPK), a member of the DAPK family, in regulating DAPK1 expression in response to IFNs. We hypothesize that ZIPK regulates DAPK1 expression by modulating the phosphorylation of C/EBP-binding domain of Med1. This aspect will be investigated in specific aim 3. We will evaluate the critical relevance of these factors to DAPK1 and autophagy using RNAi, knockout mice, protein-interactions, ChIP assays, mutagenesis and transcriptional analyses. Knowledge gained from an understanding these pathways, will not only define the critical regulators of DAPK1, but also will provide indicators into how a loss of DAPK1 can occur. We will investigate the relevance of these pathways to human CLL, a disease in which dapk1 appears to play an important tumor suppressive role. These in turn will allow a better design of therapeutics to combat tumor progression and metastasis. PUBLIC HEALTH RELEVANCE: Tumor metastasis occurs due to a loss of certain critical genes and a suppression of corresponding biological processes. Studies proposed in this application will investigate the regulation of an anti-metastatic gene, DAPK1, which is critical for tumor suppression and autophagic responses.

描述（由申请人提供）：细胞因子干扰素家族对于促进多种生理过程，例如抗病毒，抗肿瘤和免疫反应至关重要。它们是用于治疗多种癌症，病毒疾病和神经退行性疾病的临床用途。通过与其他细胞因子相互作用，IFN会形成一个控制肿瘤细胞生长和针对病原体的宿主防御的大型细胞间信号分子网络。以前，我们已经确定了一种新型的IFN调节元素及其同源转录因子。一种这样的蛋白质是CCAAT/增强子结合蛋白-BETA（C/EBP-2），这是已知的转录因子，可调节细胞分化，能量代谢，免疫反应，肿瘤生长和凋亡。我们实验室中的基因表达微阵列分析确定了几个IFN调节基因，它们的表达需要C/EBP-2。其中之一是死亡相关蛋白激酶1（DAPK1），这是凋亡，细胞周期和转移的重要调节剂。 DAPK1基因的表达经常在几种人类癌症中丧失。 DAPK1还调节自噬（一种新型的死亡形式），这对于去除受损细胞器，与细胞内病原体，抗原表现和抑制肿瘤至关重要。有趣的是，小鼠中C/EBP-2基因的丧失会导致许多这些缺陷。在最后一个资金期间，我们在调节DAPK1中表现出C/EBP-2的核心作用。在初步研究中，我们表明DAPK1的表达需要其他交易因素。值得注意的是，激活转录因子6（ATF6）是内质网依赖性应力反应的关键调节剂，似乎调节了DAPK1表达。我们提出，ATF6和C/EBP-2之间的直接相互作用会导致DAPK1表达和通过自噬抑制，其中MAP激酶凋亡 - 刺激激酶1（ASK1）提供了关键的信号输入。在该提案的特定目的1中，我们将研究C/EBP-2和ATF6如何合作以上调DAPK1表达。在特定的目标2中，我们将研究Ask1如何通过ATF6控制DAPK1的表达。增强子结合的转录因子（TFS）使用转录共激活因子促进转录。其中一个由由多种蛋白质组成的分子桥介体传达了从TFS到TATA-Box的一般转录因子复合物的转录信号。在上一个资金期间的研究还确定了Med1，Med1是C/EBP-2的IFN诱导的结合伴侣的主要亚基。我们为DAPK家族的Zip-激酶（Zipk）（Zipk）（Zipk）（Zipk）（Zipk）（Zipk）（Zipk）（ZIPK）（ZIPK）的成员进行了初步证据，在调节对IFNS响应的DAPK1表达方面。我们假设Zipk通过调节Med1的C/EBP结合结构域的磷酸化来调节DAPK1的表达。这方面将在特定目标3中进行研究。我们将使用RNAi，基因敲除小鼠，蛋白质相互作用，芯片分析，诱变和转录分析评估这些因素与DAPK1和自噬的关键相关性。从理解这些途径中获得的知识不仅将定义DAPK1的关键调节剂，而且还将为如何发生DAPK1的损失提供指标。我们将研究这些途径与人类CLL的相关性，这种疾病在这种疾病中似乎起着重要的肿瘤抑制作用。反过来，这些将允许更好地设计治疗疗法，以打击肿瘤的进展和转移。 公共卫生相关性：肿瘤转移是由于某些关键基因的丧失和相应生物学过程的抑制而发生的。本应用中提出的研究将研究抗转移基因DAPK1的调节，这对于抑制肿瘤和自噬反应至关重要。