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

• 批准号: 7933646
• 负责人: JAY YANG
• 金额: $ 29.4万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 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/7933646
• 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

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：工程原代 T 淋巴细胞的体内过继转移，并检查小鼠盲肠结扎穿刺模型中对生存的影响。 败血症。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，以及在脓毒症小鼠模型中检查的整个动物水平的直接生物学相关性。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。