Molecular Control of Cytokine Withdrawal-Induced Apoptosis in Human Lymphocytes

细胞因子撤退诱导的人淋巴细胞凋亡的分子控制

• 批准号: 7291157
• 负责人: JOAO B OLIVEIRA FILHO
• 金额: $ 5.08万
• 依托单位: FUNDACAO FACULDADE DE MEDICINA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7291157
• 关键词: 3-methyladenine; Activated Lymphocyte; Apoptosis; Apoptotic; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Autophagocytosis; BAD gene; BAX gene; BCL2L11 gene; BIM Bcl-2-binding protein; Bad protein; Bax protein; Binding; Bone Marrow; Boxing; Cell Death; Cell Line; Cell Survival; Cells; Cessation of life; Chemicals; Clinical; Confocal Microscopy; Cytochromes; Cytotoxic agent; DLEC1 gene; Data; Defect; Disease; Dyes; Enlargement of lymph nodes; Epithelial Cells; Evaluation; Extracellular Signal Regulated Kinases; Family; Family member; Flow Cytometry; Future; Genetic Screening; Goals; Hematologic Neoplasms; Human; Infection; Inflammation; Interleukin-2; Interleukin-3; Invaded; Investigation; Knock-out; Knockout Mice; Knowledge; Literature; Lymphatic Diseases; Lymphocyte; Lymphoid; MAP Kinase Gene; MG132; MLLT7 gene; Malignant - descriptor; Malignant Neoplasms; Malignant lymphoid neoplasm; Measures; Mediating; Membrane Potentials; Microtubules; Mitochondria; Mitogen-Activated Protein Kinases; Modeling; Molecular; Monitor; Mus; Mutation; N-ras Genes; NRAS gene; Neurons; Normal Cell; Osteoclasts; Pathway interactions; Patients; Pattern; Peripheral Blood Lymphocyte; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Plasmids; Play; Polymerase Chain Reaction; Post-Translational Regulation; Prevention; Principal Investigator; Process; Proteasome Inhibitor; Protein Structure Initiative; Proteins; Proto-Oncogene Proteins c-jun; Radiation; Relative (related person); Research Personnel; Resistance; Role; Signal Transduction; Small Interfering RNA; Splenomegaly; Staining method; Stains; Syndrome; System; Testing; Tissues; Transcriptional Activation; Transcriptional Regulation; Transfection; Transmission Electron Microscopy; United States National Institutes of Health; Up-Regulation; Western Blotting; Withdrawal; Work; apoptosis in lymphocytes; autoimmune lymphoproliferative syndrome; base; c JUN kinase; cell type; cohort; cytokine; design; dynein light chain; falls; forkhead protein; in vitro Assay; inhibitor/antagonist; interest; jun Oncogene; kinase inhibitor; knock-down; lactacystin; multicatalytic endopeptidase complex; mutant; p21 N-Ras Protein; pathogen; programs; research study; stem; therapeutic target; therapy design; transcription factor; wortmannin

DESCRIPTION (provided by applicant): After clearance of an infection the level of several cytokines essential for the survival of the responding activated lymphocytes falls, triggering lymphocyte death. Our long-term goal is to elucidate the molecular control of apoptosis triggered by cytokine withdrawal in human lymphocytes. A better understanding of the mechanisms regulating this form of apoptosis in lymphocytes may reveal strategies for designing new therapies for autoimmune diseases and hematologic malignancies. In fact, using an approach similar to the proposed in this project we have recently identified a new genetic defect causing a syndrome of lymphoid accumulation, autoimmunity and cancer. Lymphocytes from BIM, PUMA, and BAD knockout mice are resistant to apoptosis induced by cytokine withdrawal. Multiple pathways regulate BIM in different cell types, and the mechanism regulating this protein in human lymphocytes is unknown. BIM is inactivated by binding to dynein light chains in epithelial cells. Transcription factors of the forkhead box o (FOXO) family transcriptionally regulate BIM in several cell lines. The extracellular-signal regulated kinase (ERK)/MAPK pathway can control the baseline levels of BIM, and the c-JUN amino-terminal kinase (JNK) pathway controls the levels of BIM transcriptionally and post-translationally in neurons and other cell types. In addition, autophagy also seems to protect cell lines from cytokine withdrawal-induced cell death. Based on these observations, our specific aims are to: 1) Characterize the mechanism(s) regulating the levels and activation of BIM after cytokine withdrawal. We will investigate the role of the FOXO family, JNK and ERK kinase pathaways on the transcripitional and post- translational regulation of BIM and during IL-2 withdrawal, by the use of siRNA and chemical inhibitors. 2) Determine the relative importance of PUMA and BAD for the initiation of cell death following cytokine withdrawal in human lymphocytes. We will use siRNA to knock down each of these proteins and evaluate the impact on apoptosis, cytochrome C release, and BAX activation induced by cytokine withdrawal. 3) Determine the role of autophagy as an anti-apoptotic mechanism after IL-2 withdrawal in primary lymphocytes. The presence of autophagy after cytokine withdrawal will be measured and drug treatments used to block autophagy. The impact of these manipulations on cell death induced by IL-2 withdrawal will be quantified. Our long-term goal is to elucidate the molecular control of apoptosis triggered by cytokine withdrawal in human lymphocytes. A better understanding of the mechanisms regulating this form of apoptosis in lymphocytes may reveal strategies for designing new therapies for autoimmune diseases and hematologic

描述（由申请方提供）：感染清除后，应答活化淋巴细胞存活所必需的几种细胞因子水平福尔斯，触发淋巴细胞死亡。我们的长期目标是阐明人类淋巴细胞中细胞因子撤退引发的细胞凋亡的分子控制。更好地理解调节淋巴细胞中这种形式的凋亡的机制可能揭示设计自身免疫性疾病和血液恶性肿瘤的新疗法的策略。事实上，使用与该项目中提出的方法类似的方法，我们最近发现了一种新的遗传缺陷，导致淋巴积聚，自身免疫和癌症综合征。来自BIM、CD 4A和BAD敲除小鼠的淋巴细胞对细胞因子戒断诱导的细胞凋亡具有抗性。多种途径调节不同细胞类型中的BIM，调节人类淋巴细胞中这种蛋白质的机制尚不清楚。BIM通过与上皮细胞中的动力蛋白轻链结合而失活。FOXO家族的转录因子在几种细胞系中转录调节BIM。细胞外信号调节激酶（ERK）/MAPK途径可以控制BIM的基线水平，c-JUN氨基末端激酶（JNK）途径控制神经元和其他细胞类型中BIM的转录和转录后水平。此外，自噬似乎也保护细胞系免受细胞因子撤出诱导的细胞死亡。基于这些观察，我们的具体目标是：1）表征在细胞因子撤除后调节BIM的水平和活化的机制。我们将通过使用siRNA和化学抑制剂研究FOXO家族、JNK和ERK激酶通路对BIM的转录和翻译后调节以及在IL-2戒断期间的作用。2)确定人淋巴细胞中细胞因子撤出后，CD 4A和BAD对细胞死亡起始的相对重要性。我们将使用siRNA敲除这些蛋白质中的每一种，并评估对细胞凋亡、细胞色素C释放和细胞因子戒断诱导的BAX激活的影响。3)确定自噬作为原代淋巴细胞中IL-2撤除后的抗凋亡机制的作用。将测量细胞因子撤除后自噬的存在，并使用药物治疗来阻断自噬。将定量这些操作对IL-2戒断诱导的细胞死亡的影响。我们的长期目标是阐明人类淋巴细胞中细胞因子撤退引发的细胞凋亡的分子控制。更好地理解调节淋巴细胞中这种形式的凋亡的机制可能会揭示设计自身免疫性疾病和血液病新疗法的策略。