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: 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
• 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

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)：败血症是一种主要的医疗保健问题，仅在美国一年的发病率就超过75万例，给人类带来巨大的痛苦和国家医疗资源的损失。导致这种疾病高死亡率的基本机制尚不清楚，但积累的数据表明，早期T淋巴细胞凋亡和随后的低免疫阶段在生物体死亡中起着关键作用。如果丢失的T淋巴细胞能够被补充，生物体可能能够抵抗感染，提高存活率。质膜Kv1.3钾通道在T淋巴细胞活化中的关键作用已被认识到一段时间。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器。我们建议通过抑制线粒体Kv1.3钾通道来创造相对抵抗凋亡的T淋巴细胞，并探索过继转移这些基因工程T淋巴细胞作为一种治疗脓毒症的新方法。其工作假设是“抑制淋巴细胞线粒体Kv1.3离子通道可防止细胞凋亡，过继转移抗凋亡的工程初级T淋巴细胞将提高脓毒症患者的存活率”。目的1：研究线粒体Kv1.3在T淋巴细胞凋亡中的作用，目的2：研究线粒体Kv1.3调控细胞凋亡的机制，目的3：体内过继转移原代工程T淋巴细胞，观察其对小鼠盲肠结扎穿孔脓毒症模型存活的影响。这一创新假说将使用广泛的最先进技术进行验证，包括基于慢病毒的原代T淋巴细胞体外工程、体外和体外细胞凋亡评估、各种分子结构的细胞器特异性表达，以及在脓毒症小鼠模型中检验整个动物水平的直接生物学相关性。强调从机制上了解线粒体Kv1.3在调节细胞凋亡中的作用，将这一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，组成了执行这一多方面项目所需的独特的专业知识集合。