IN VITRO DIAGNOSTIC TEST FOR LITHIUM-SENSITIVE BIPOLAR DISORDER

锂敏感双相情感障碍的体外诊断测试

• 批准号: 8173613
• 负责人: CHIAKI FUKUHARA
• 金额: $ 17.2万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173613
• 关键词: Acute; Agreement; Biological Assay; Bioluminescence; Bipolar Disorder; Cell Line; Cells; Circadian Rhythms; Clinical; Computer Retrieval of Information on Scientific Projects Database; Cultured Cells; Diagnostic tests; Fibroblasts; Funding; Goals; Grant; Human; In Vitro; Institution; Length; Lithium; Luc Gene; Luciferases; Methods; Monitor; Participant; Patients; Peripheral; Publishing; Recruitment Activity; Reporter; Research; Research Personnel; Resources; Source; System; Testing; Therapeutic; Time; United States National Institutes of Health; circadian pacemaker; promoter; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A goal of our present proposal is to establish an assay system by which circadian rhythms can be monitored using human fibroblasts with a circadian reporter and real time bioluminescence monitoring, and to test our hypothesis that clinical response to lithium can be predicted using human peripheral cells. b. Studies and Results. The goal of the first Aim is to apply in vitro circadian rhythm recording method using human primary fibroblasts. So far eleven healthy participants were recruited and six fibroblast cell lines have been established. After cultivation and expansion, the circadian clock Bmal1 promoter-luciferase gene was delivered to fibroblasts, and Bmal1-luciferase cell lines were established. We optimized culture and recording conditions. Robust and sustained circadian rhythms have now been recorded in these human fibroblast cell lines. Average period length was 24.60 ¿ 0.16 h (N=5). The results were in agreement with those previously published by Dr. Brown and his colleagues (Brown et al., 2005) and, therefore, we have completed the Aim 1. In Aim 2 we plan to examine the effects of lithium on circadian rhythm parameters in fibroblasts obtained from lithium responders and non-responders. We have been optimizing lithium treatment conditions to be applied to bipolar disorder patients' cells. Since high lithium concentrations were used to test the effects of lithium on the period length in previous studies, we tested 3, 5, 7, and 10 mM lithium stimulation on the period length. At the same time, we want to test the effects of lithium at therapeutic concentrations, e.g., 1 mM. Since acute 1mM lithium stimulation did not show significant impact, we have been culturing cells in the presence of l mM lithium for one or four weeks before rhythm recording. We chose four weeks as well because the clinical lithium sensitivity is first determined four weeks after lithium therapy.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 我们目前提案的目标是建立一种检测系统，通过该系统可以使用具有昼夜节律报告器和实时生物发光监测的人类成纤维细胞来监测昼夜节律，并检验我们的假设，即可以使用人类外周细胞来预测对锂的临床反应。 b. 研究和结果。 第一个目标是利用人原代成纤维细胞应用体外昼夜节律记录方法，目前已招募11名健康参与者，并建立了6个成纤维细胞系，经过培养和扩增，获得了生物钟Bmal1启动子-荧光素酶基因。递送至成纤维细胞，并建立了 Bmal1-荧光素酶细胞系，并优化了这些人的培养和记录条件。成纤维细胞系的平均周期长度为 24.60 ¿ 0.16 小时（N=5）。结果与 Brown 博士及其同事之前发表的结果一致（Brown 等人，2005），因此，我们完成了目标 1。 在目标 2 中，我们计划检查锂对从锂反应者和非反应者获得的成纤维细胞的昼夜节律参数的影响，因为使用高浓度的锂，我们一直在优化应用于双相情感障碍患者细胞的锂治疗条件。在之前的研究中测试锂对经期长度的影响，我们测试了3、5、7和10 mM锂刺激对经期长度的影响。现在，我们想要测试治疗浓度（例如 1 mM）锂的效果，由于急性 1 mM 锂刺激没有显示出显着影响，因此我们在节律记录之前在 1 mM 锂存在下培养细胞一到四个星期。我们也选择了 4 周，因为临床锂敏感性是在锂治疗后 4 周首先确定的。