Mitochondrial Kv1.3 potassium channels and lymphocyte apoptosis in sepsis

脓毒症中线粒体 Kv1.3 钾通道与淋巴细胞凋亡

基本信息

批准号：
7692893
负责人：
JAY YANG
金额：
$ 29.7万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-30 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7692893
关键词：
Abbreviations Adoptive Transfer Animals Antigen Presentation Apoptosis Apoptotic Area Arts Basic Science Biological Biology C-terminal Carbonyl Cyanide m-Chlorophenyl Hydrazone Cardiovascular system Caspase Caspase Inhibitor Cell Death Cell Therapy Cell membrane Cells Cellular biology Cessation of life Clinical Collaborations Collection Complex Cycloheximide Cytotoxic T-Lymphocytes Data Development Disease Electrophysiology (science)Embryo Endoplasmic Reticulum Engineering Environment Epitopes Equipment Fertilization Future Glioma Healthcare Healthcare Systems Hemagglutinin Human Human Resources Hydrogen Peroxide Immune Immune response Immunoblotting Immunologist Immunoprecipitation Immunosuppression In Vitro Incidence Infection Inflammatory Response Intensive Care Interleukins Invaded Investigation Ion Channel Japan Kidney Knowledge Kv1.3 potassium channel Laboratories Letters Ligation Literature Lymphocyte Lymphocyte Transfusion Malignant Neoplasms Mediating Membrane Potentials Memorial Sloan-Kettering Cancer Center Methodology Mitochondria Modeling Molecular Molecular Biology Mus Organelles Organism Pathology Pathway interactions Patients Phase Play Postdoctoral Fellow Potassium Channel Process Productivity Proline-Rich Domain Publications Puncture procedure RNA Receptor Protein-Tyrosine Kinases Regulation Relative (related person)Reporting Research Research Personnel Resistance Resources Role Scheme Science Scientist Sepsis Signal Transduction Small Interfering RNA Subfamily lentivirinae Survival Rate T-Lymphocyte Techniques Testing Time Training Translations Tumor Necrosis Factor Ligand Superfamily Member 6 Tumor Necrosis Factor-alpha Tumor Necrosis Factors United States Universities Voltage-Dependent Anion Channel Voltage-Gated Potassium Channel Work Yang apoptosis in lymphocytes base cost endoplasmic reticulum stress enhanced green fluorescent protein experience fighting in vivo innovation interest mitochondrial membrane mortality mouse model new therapeutic target novel patch clamp pre-clinical prevent pro-apoptotic protein professor research study response small hairpin RNA success synergism

项目摘要

DESCRIPTION (provided by applicant): Sepsis is a major health care problem with incidence exceeding 750,000 cases a year in the United States alone with an enormous cost to human suffering and national healthcare resources. The basic mechanisms responsible for the high mortality rate of this disease remain unknown but cumulative data points to a key role of early T lymphocyte apoptosis and the subsequent development of a hypo-immune phase in the death of the organism. If the lost T lymphocytes can be replenished the organism may be able to fight the infection and increase the survival rate. The key role of plasma membrane Kv1.3 potassium channels in the activation of T lymphocytes has been recognized for some time. We discovered that this ion channel also present in the mitochondria is a powerful regulator of T lymphocyte apoptosis. We propose to create T lymphocytes rendered relative resistance to apoptosis through inhibition of the mitochondrial Kv1.3 potassium channels and explore adoptive transfer of these genetically engineered T lymphocytes as a novel treatment for sepsis. The working hypothesis is "inhibition of lymphocyte mitochondrial Kv1.3 ion channels prevents apoptosis and adoptive transfer of apoptosis-resistant engineered primary T lymphocytes will enhance survival in sepsis". The three specific aims of the proposal are: Aim 1: Examination of the role of mitochondrial Kv1.3 in T lymphocyte apoptosis, Aim 2: Examination of the mechanisms of mitochondrial Kv1.3 regulation of apoptosis, and Aim 3: In vivo adoptive transfer of engineered primary T lymphocytes and examination of the effect on survival in the murine cecal-ligation-puncture model of sepsis. The innovative hypothesis will be tested using a wide range state-of-art techniques including lentivirus-based ex vivo engineering of primary T lymphocytes, ex vivo and in vitro assessment of apoptosis, organelle-specific expression of various molecular constructs, and the direct biological relevance at the whole animal level examined in a mouse model of sepsis. The emphasis placed on gaining a mechanistic understanding of the role of mitochondrial Kv1.3 in the regulation of apoptosis distinguishes this proposal from a purely pre-clinical phenomenological research. The research team, along with the consultants, comprises a unique collection of expertise necessary to execute this multi-faceted project.

描述（由申请人提供）：脓毒症是一个主要的医疗保健问题，仅在美国每年的发病率就超过 750,000 例，给人类痛苦和国家医疗资源造成巨大损失。造成这种疾病高死亡率的基本机制仍然未知，但累积的数据表明早期 T 淋巴细胞凋亡和随后的免疫低下阶段在生物体死亡中发挥着关键作用。如果丢失的 T 淋巴细胞能够得到补充，生物体或许能够抵抗感染并提高存活率。一段时间以来，人们已经认识到质膜 Kv1.3 钾通道在 T 淋巴细胞激活中的关键作用。我们发现这种离子通道也存在于线粒体中，是 T 淋巴细胞凋亡的强大调节剂。我们建议通过抑制线粒体 Kv1.3 钾通道来创建对细胞凋亡具有相对抵抗力的 T 淋巴细胞，并探索这些基因工程 T 淋巴细胞的过继转移作为脓毒症的新型治疗方法。工作假设是“抑制淋巴细胞线粒体 Kv1.3 离子通道可防止细胞凋亡，抗凋亡工程原代 T 淋巴细胞的过继转移将提高脓毒症患者的存活率”。该提案的三个具体目标是：目标 1：检查线粒体 Kv1.3 在 T 淋巴细胞凋亡中的作用，目标 2：检查线粒体 Kv1.3 调节细胞凋亡的机制，目标 3：体内过继转移工程原代 T 淋巴细胞的改造以及对小鼠脓毒症盲肠结扎穿刺模型中生存的影响的检查。该创新假设将使用广泛的最先进技术进行测试，包括基于慢病毒的原代 T 淋巴细胞离体工程、细胞凋亡的离体和体外评估、各种分子构建体的细胞器特异性表达以及直接生物学研究。在脓毒症小鼠模型中检查了整个动物水平的相关性。重点在于从机制上理解线粒体 Kv1.3 在细胞凋亡调节中的作用，使该提议与纯粹的临床前现象学研究区分开来。研究团队与顾问一起，拥有执行这一多方面项目所需的独特专业知识。