FAM26F function and role in macrophages

FAM26F 在巨噬细胞中的功能和作用

• 批准号: 10449780
• 负责人: William A Coetzee
• 金额: $ 16.95万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10449780
• 关键词: Affect; Atherosclerosis; Autoimmune Diseases; Automobile Driving; Biochemical; Biophysics; CRISPR/Cas technology; Cartilage; Cell Culture Techniques; Cells; Cellular Assay; Characteristics; Chondrocytes; Clinical; Data; Disease; Disease Progression; Disease model; Drug Targeting; FDA approved; Family; Family member; Genes; Goals; Hematological Disease; Histologic; Homidium Bromide; Immune; Immune System Diseases; Immune system; Immunity; Immunologist; Immunology; Immunotherapy; Impairment; Inflammasome; Inflammation; Inflammatory; Innate Immune System; Integral Membrane Protein; Interleukin-1 beta; Ion Channel; Life; Link; Lymphocyte; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Measures; Mediating; Membrane; Methods; Molecular Weight; Mus; Muscle Fibers; Myocardium; NK Cell Activation; Names; Natural Immunity; Natural Killer Cells; Neurons; Nucleotides; Osteoclasts; Pathology; Patients; Physiological Processes; Play; Production; Protein Family; Proteins; Proteomics; Publishing; Pump; Regulation; Research; Resolution; Resources; Rheumatoid Arthritis; Rodent Model; Role; Structure; Synovitis; TNF gene; Testing; Therapeutic; bone erosion; cytokine; experimental study; joint destruction; joint inflammation; macrophage; member; mouse model; neurotransmitter release; novel; overexpression; selective expression; small molecule; therapeutic target; uptake

Abstract Ion channels, transporters, exchangers and pumps (here called ICTs) control most aspects of life. They have been extensively characterized in excitable cells, such as neurons, where they control neurotransmitter release and other functions as well as cardiac and skeletal muscle cells, where they regulate the strength of contraction. In most non-excitable cells, and specifically in immune cells, ICTs are only scantly characterized. We have taken a global approach to analyze the expression of more than 600 ICTs in immune cells, and observed that one particular putative ICT, named FAM26F, is highly and specifically expressed in proinflammatory macrophages compared to other immune cells. FAM26F is an understudied ion channel that belongs to a small family of proteins (FAM26A to F). The best characterized FAM26 family member is FAM26C. It functions as an ion channel under certain experimental conditions and has been described to promote ATP release from cells. Recently published low resolution structures of FAM26F show that it, like FAM26C, also forms a pore and may act as an ion channel. Our preliminary data to date, however, did not detect evidence for channel function of FAM26F when overexpressed in mammalian cells. The most likely reason is that FAM26F needs other subunits or associated proteins to function as a channel, which is characteristic of many ion channels. We hypothesize that while overexpression of FAM26F may not be sufficient to generate a functional channel, and proinflammatory macrophages express not only FAM26F but also the other proteins required to form a channel. To test this hypothesis, we will in Aim 1 directly measure putative functions of FAM26F such as channel currents and the transport of low molecular weight molecules. In a parallel approach, we will use biochemical methods and mass spectrometry to identify the proteins that associate with FAM26F in macrophages. In Aim 2, we will elucidate the role of FAM26F in macrophage function and inflammation. Our preliminary data have shown that deletion of FAM26F impairs certain proinflammatory functions of macrophages such as the release of cytokines. These immune messenger proteins are critical for disease progression in the autoimmune disease rheumatoid arthritis, where they drive joint inflammation and destruction. We will therefore use macrophages and mice in which FAM26F has been deleted to determine the effects of FAM26F on macrophage function in cell culture settings and in mouse models of rheumatoid arthritis. This multi-PI R03 proposal by Drs. Stefan Feske and William Coetzee brings together their unique expertise to better understand the roles of channels, and in particular FAM26F, in innate immunity. There are currently no FDA approved drugs that target ion channels for immunological disorders and the completion of the proposed studies takes us an important step in the direction of this missed therapeutic opportunity.

抽象的 离子通道、转运蛋白、交换器和泵（此处称为 ICT）控制着生命的大部分方面。他们有 在可兴奋细胞（例如神经元）中被广泛表征，它们控制神经递质的释放 和其他功能以及心肌细胞和骨骼肌细胞，它们调节收缩的强度。 在大多数不可兴奋的细胞中，特别是在免疫细胞中，ICT 的特征很少。我们已经采取了 一种分析免疫细胞中 600 多种 ICT 表达的全球方法，并观察到 特定的假定 ICT，名为 FAM26F，在促炎巨噬细胞中高度特异性表达 与其他免疫细胞相比。 FAM26F 是一种正在研究的离子通道，属于一个小家族 蛋白质（FAM26A 至 F）。最具特征的 FAM26 家族成员是 FAM26C。它起到离子通道的作用 在某些实验条件下，并已被描述为促进细胞中 ATP 的释放。最近 已发表的 FAM26F 的低分辨率结构表明，它与 FAM26C 一样，也形成孔，并可能充当 离子通道。然而，我们迄今为止的初步数据并未发现 FAM26F 通道功能的证据 当在哺乳动物细胞中过度表达时。最可能的原因是 FAM26F 需要其他亚基或 相关蛋白质充当通道，这是许多离子通道的特征。我们假设 而 FAM26F 的过度表达可能不足以产生功能性通道，并且促炎性通道 巨噬细胞不仅表达 FAM26F，还表达形成通道所需的其他蛋白质。为了测试这个 假设，我们将在目标 1 中直接测量 FAM26F 的假定功能，例如通道电流和 低分子量分子的运输。在并行方法中，我们将使用生化方法和质量 光谱法来鉴定与巨噬细胞中 FAM26F 相关的蛋白质。在目标 2 中，我们将阐明 FAM26F 在巨噬细胞功能和炎症中的作用。我们的初步数据表明，删除 FAM26F 损害巨噬细胞的某些促炎功能，例如细胞因子的释放。这些 免疫信使蛋白对于自身免疫性疾病类风湿关节炎的疾病进展至关重要， 它们会导致关节炎症和破坏。因此，我们将使用巨噬细胞和小鼠，其中 FAM26F 已被删除，以确定 FAM26F 对细胞培养环境中巨噬细胞功能的影响 以及类风湿性关节炎的小鼠模型。博士的这个多 PI R03 提案。斯特凡·费斯克和威廉 Coetzee 汇集了他们独特的专业知识，以更好地理解渠道的作用，特别是 FAM26F，先天免疫。目前尚无 FDA 批准的针对离子通道的药物 免疫性疾病和拟议研究的完成使我们朝着这个方向迈出了重要的一步 这次错过的治疗机会。