Clinical Studies Of Abnormal Host Defense

宿主防御异常的临床研究

基本信息

批准号：
9161429
负责人：
JOHN I GALLIN
金额：
$ 21.55万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9161429
关键词：
Age Arterial Fatty Streak Atherosclerosis Base Sequence Blood Circulation CXCR4 gene Calcium Carotid Arteries Cells Chemicals Chronic Granulomatous Disease Clinical Clinical Research Collaborations Complex Coronary artery Data Defect Deposition Development Disease Doctor of Philosophy Family member Genetic study Goals Host Defense Host Defense Mechanism Human Hypoxia ITGB2 gene Image Immune System Diseases Immunodominant Antigens Immunologic Deficiency Syndromes Individual Infection Infectious Agent Inflammation Investigation Laboratories Laboratory Study Link Magnetic Resonance Imaging Manuscripts Measures Metabolism Methanol Methodology Molecular Molecular Diagnosis Monitor Mutation NADP NADPH Oxidase Natural History Normal Cell Nucleic acid sequencing Organism Oxidases Oxidoreductase Oxygen Paper Pathogenesis Patients Phagocytes Pilot Projects Process Production Protocols documentation Publishing Reactive Oxygen Species Role Sampling Therapeutic Translational Research United States National Institutes of Health cell growth regulation cohort congenital immunodeficiency digital gene therapy genetic immunotherapy high throughput screening human subject immune function improved induced pluripotent stem cell inhibitor/antagonist innate immune function metabolomics neutrophil neutrophil cytosol factor 40K new technology novel novel strategies novel therapeutics pathogen pre-clinical screening sex small molecule

项目摘要

1) Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by mutations in the multicomponent NADPH oxidase (phagocyte oxidase, NOX2) complex. During the past FY, through collaboration with the Neutrophil Monitoring Laboratory (NML) managed by Douglas Kuhns, PhD ( Leidos, Inc.), we provided molecular diagnoses using immunodetection of components of the NADPH oxidase for 4 p47phox-deficient- and 12 gp91phox-deficient subjects. Nucleic acid sequencing determined the mutations in 33 patients and family members including the rarest form, p40phox deficiency. The NML has also provided molecular diagnosis of other patients with immunodeficiencies due to mutations in CXCR4, ITGB2, WDR1, and PADI4. Dr. Kuhns has developed a novel approach to determining p47 mutations relying on digital droplet PCR (manuscript in prep). During this FY, we collaborated with the Genetic Immunotherapy Section of the LHD to demonstrate novel gene therapy approaches to correct induced pluripotent stem cells from CGD patients (Merling et al., Molecular Therapy 2015). We are currently collaborating with the Human Immunological Disease Unit of the LHD to evaluate innate immune function in phagocytes from several patients. 2) Our group continues its clinical and laboratory studies of the emerging Gram-negative CGD pathogen, Granulibacter bethesdensis. During FY15, we screened 11 CGD samples and 15 normal samples for seropositivity toward G. bethesdensis methanol dehydrogenase, an immunodominant antigen. We continue to monitor seropositivity in culture-confirmed patients to evaluate our hypothesis that this organism can establish persistent, clinically unapparent infections. 3) During FY15, we published a paper describing data from NIH Protocol #10-I-0029 Non-invasive Assessment of Atherosclerosis in Patients with CGD and other Disorders of the Immune System (current total = 85 subjects). Atherosclerosis is caused, in part, by inflammation in the vasculature and over production of reactive oxygen species (ROS) has been implicated in pathogenesis of this disease. We hypothesized that CGD patients, who have deficient production of reactive oxygen species by their phagocytes and other cells, may be protected from developing atherosclerosis. The primary endpoint of this study was the assessment of atherosclerotic plaque formation/calcium deposition in the carotid and coronary arteries by CT, MRI and other imaging methodologies, in these and other patients with in-born disorders of immune function. We found significantly smaller carotid vessel wall volumes, a pre-clinical measure of atherosclerosis, in CGD patients compared to age- and sex-matched control subjects. This finding was published in Circulation (Sibley et al.). We continue to study patients on this protocol and have focused on carriers of X-linked CGD reasoning that these otherwise healthy individuals should have subnormal amounts of ROS and that this may be protective in this somewhat older cohort of subjects. This current effort will prove the hypothesis that lower levels of ROS production are protective although the situation in the X-linked CGD carriers differs from the CGD patients in that some cells are normal and some are abnormal. Simultaneously, we are collaborating with the National Center for Advancing Translational Sciences (NCATS), involving the development of high throughput assays for NADPH oxidase activity to permit large-scale screening of chemical inhibitors of NOX2. (4) In normal PMN, activation of NOX2 results in a rapid (within seconds to minutes) depletion of NADPH and hypoxia. Given the central roles of oxygen and NAPDH in metabolism, we compared the metabolomes of PMN before and after activation of NOX2 in a pilot study and found unexpected alterations in PMN metabolism. We are repeating these studies in PMN from patients with CGD to determine the exact role of ROS in these processes.

1）慢性肉芽肿性疾病（CGD）是由多组分NADPH氧化酶（吞噬细胞氧化酶，NOX2）复合物中突变引起的主要免疫缺陷。在过去的FY中，通过与道格拉斯·库恩斯（Douglas Kuhns，PhD）（Leidos，Inc。）管理的中性粒细胞监测实验室（NML）合作，我们使用NADPH氧化酶的免疫进行了分子诊断，以4 p47phox-deceigient-declox-deceient-declox-declox-declos-dec91phox-Phox-Phox-ficeentects进行分子诊断。核酸测序确定了33名患者和家庭成员的突变，包括最稀有的形式P40Phox缺乏症。 NML还提供了由于CXCR4，ITGB2，WDR1和PADI4突变引起的其他免疫缺陷患者的分子诊断。库恩斯博士开发了一种新的方法来确定依靠数字液滴PCR（PREP中的手稿）的P47突变。在此FY期间，我们与LHD的遗传免疫疗法部分合作，展示了新型的基因治疗方法，以纠正CGD患者的多能干细胞（Merling等，Merling等，Molecular Therapy 2015）。我们目前正在与LHD的人类免疫疾病单位合作，以评估几名患者的吞噬细胞中的先天免疫功能。 2）我们的小组继续对新兴的革兰氏阴性CGD病原体，肉芽肿杆菌贝丝斯氏菌进行临床和实验室研究。在2015财年期间，我们筛选了11个CGD样品和15个正常样品，以对贝塞斯丁菌甲醇脱氢酶（一种免疫抗原）的血清阳性。我们继续监测培养确认患者中的血清阳性，以评估我们的假设，即这种生物可以建立持续的临床上不明显的感染。 3）在2015财年期间，我们发表了一篇论文，描述了NIH方案＃10-I-0029的数据，对CGD患者的动脉粥样硬化和免疫系统其他疾病的动脉粥样硬化评估（当前总数= 85名受试者）。动脉粥样硬化是由于脉管系统的炎症而引起的，而活性氧（ROS）的产生与该疾病的发病机理有关。我们假设CGD患者可以保护其吞噬细胞和其他细胞产生活性氧，但可以保护不受动脉粥样硬化的影响。这项研究的主要终点是评估了CT，MRI和其他成像方法论，在这些和其他患有免疫功能疾病的患者中，CT，MRI和其他成像方法对颈动脉和冠状动脉动脉的钙沉积进行了评估。与年龄和性别匹配的对照组相比，我们发现CGD患者的颈动脉血管壁量显着较小，这是动脉粥样硬化的临床前测量。这一发现发表在循环中（Sibley等人）。我们继续根据该方案研究患者，并专注于X连锁CGD的载体，即这些健康的个体应该具有一定数量的ROS，并且在这一较老的受试者中，这可能具有保护性。目前的努力将证明较低水平的ROS产生是保护性的，尽管X连锁的CGD载体的情况与CGD患者不同，因为某些细胞是正常的，有些细胞异常。同时，我们正在与国家前进的转化科学中心（NCAT）合作，涉及开发NADPH氧化酶活性的高吞吐量测定，以允许对NOX2的化学抑制剂进行大规模筛查。（4）在正常的PMN中，NOX2的激活导致NADPH和缺氧的耗竭迅速（在几秒钟内）。鉴于氧气和NAPDH在代谢中的中心作用，我们比较了NOX2激活前PMN的代谢组，并发现PMN代谢意外改变。我们正在从CGD患者的PMN中重复这些研究，以确定ROS在这些过程中的确切作用。