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：
7563485
负责人：
JAY YANG
金额：
$ 32.06万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
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 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7563485
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 Localized 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 Range Rate Receptor Protein-Tyrosine Kinases Regulation Relative (related person)Reporting Research Research Personnel Resistance Resources Role Scheme Science Scientist Sepsis Signal Transduction Small Interfering RNA Stress Subfamily lentivirinae Survival Rate T-Lymphocyte Techniques Testing Therapeutic immunosuppression Time Training Translations Tumor Necrosis Factor Ligand Superfamily Member 6 Tumor Necrosis Factor-alpha Tumor Necrosis Factors United States Universities VDAC1 gene Voltage-Dependent Anion Channel Voltage-Gated Potassium Channel Work Yang apoptosis in lymphocytes base cost enhanced green fluorescent protein experience fighting human TNF protein in vivo innovation interest mitochondrial membrane mortality mouse model novel novel therapeutics patch clamp pre-clinical prevent pro-apoptotic protein professor research study response small hairpin RNA success synergism therapeutic target

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）：败血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，造成了人类苦难和国家医疗保健资源的巨大成本。导致该疾病高死亡率的基本机制仍然未知，但累积数据表明早期T淋巴细胞凋亡的关键作用以及随后在生物体死亡中发育中的免疫阶段的发展。如果可以补充损失的T淋巴细胞，则有机体可能能够抵抗感染并提高生存率。质膜KV1.3钾通道在T淋巴细胞激活中的关键作用已识别一段时间。我们发现该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，并探索这些基因设计的T淋巴细胞作为败血症的新型治疗方法，从而产生对凋亡的T淋巴细胞。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和抗凋亡耐药性的原发性T淋巴细胞的过继转移可以提高脓毒症的生存率”。 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败血症。创新的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发性T淋巴细胞的离体工程，EX VIVO和凋亡的体外评估，各种分子构建体的细胞器特异性表达，各种分子构建体的特异性表达，以及在Sepsis epsis epsis epsis中所检查的整个动物水平的直接生物学相关性。强调对线粒体KV1.3在凋亡调节中的作用的机械理解的重点将这一建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，包括执行这个多方面项目所需的独特专业知识集合。