IN VITRO REAL-TIME CIRCADIAN RHYTHM MEASUREMENT

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 昼夜节律的产生由大约24小时的生理调节节律模式组成，因此对表达的适当调节对维持良好的健康至关重要。昼夜节律的紊乱似乎与许多疾病有关，如季节性情感障碍、各种其他精神疾病、睡眠问题、癌症和一些代谢综合征。昼夜节律紊乱可能在其中几种疾病的发病机制中起到一定的致病作用，因为有证据表明，与昼夜节律异常产生有关的遗传因素与双相情感障碍、家族性晚期睡眠相综合征和睡眠相延迟综合征等疾病之间存在联系。尽管在人类中已经报道了昼夜节律紊乱与几种疾病之间的联系，但对于许多疾病和障碍来说，昼夜节律产生影响疾病表现的确切机制仍然未知。限制因素包括与人体昼夜节律表达的生理测量相关的技术困难。为了更好地了解患者体内昼夜节律的产生，昼夜节律紊乱在发病机制中的意义，以及评估潜在的药物治疗方法，建立一种非侵入性的、简单、快速和可靠的体外测定人体昼夜节律的方法是很重要的。最近的发现使研究人员能够证明使用外周细胞分析人类昼夜节律的可行性。到目前为止，真皮成纤维细胞是唯一可以记录到人类昼夜节律稳定的外周细胞。然而，皮肤活检在大多数临床科室并不是常规程序；它们对患者来说也是痛苦的，而且经常会留下疤痕。因此，开发一种使用其他细胞类型(即血细胞)的检测系统将是有益的。 在目前的提案中，我们的目标是开发一种非侵入性的体外测试系统，通过使用时钟基因报告程序实时监测来自人类血液样本的巨噬细胞的昼夜节律(总共31名健康受试者将被回收)。一旦建立了检测系统，这项技术将被应用于评估化学品的影响，从而测试这一检测作为未来诊断工具的可行性(总共将回收30个样本)。 该中心将用于招募健康志愿者，并为上述目的采集血液样本。每位参与者将被要求献血两次，间隔至少一个月。

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. Circadian rhythm generation consists of an approximate 24 h rhythmic pattern of physiologic regulation whereby proper regulation of the expression is critical to maintaining good health. Disturbances in circadian rhythm generation appear to be associated with many diseases such as seasonal affective disorders, a variety of other mental illnesses, sleep problems, cancer, and a number of metabolic syndromes. It is possible that circadian disruptions have some causative role in the pathogenesis of several of these diseases, as there is evidence suggesting links between genetic factors related to abnormal circadian rhythm generation and illnesses like bipolar affective disorder, familial advanced sleep phase syndrome, and delayed sleep phase syndrome. Even though associations between circadian rhythm disturbances and several diseases have been reported in humans, the precise mechanisms by which circadian rhythm generation impacts the manifestation of disease remains unknown for many illnesses and disorders. Among the limiting factors are technical difficulties related to the physiological measurement of human circadian rhythm expression. To better understand human circadian rhythm generation in patients, the implications of circadian rhythm disturbances in pathogenesis, as well as to evaluate potential pharmaceutical treatments, it is important to develop a non-invasive, simple, rapid and reliable in vitro assay to determine circadian rhythm in humans. Recent findings have enabled investigators to demonstrate the feasibility of using peripheral cells to analyze human circadian rhythm. So far, the dermal fibroblast is the only peripheral cell in which stable circadian rhythm can be recorded in humans. However, skin biopsy is not a routine procedure in most clinical departments; they also are painful for patients and often leave scars. Therefore, it would be beneficial to develop an assay system using some other cell type (i.e., a blood cell). In the present proposal, we aim to develop a non-invasive, in vitro assay system to measure human circadian rhythm in macrophages derived from human blood samples, using real-time monitoring of circadian parameters with a clock gene reporter (a total of 31 healthy subjects will be recuited). Once the assay system is established, this technique will be applied to evaluate the effects of chemicals, thereby testing the feasibility of this assay as a future diagnostic tool (a total of 30 paritipants will be recuited). The CRC will be utilized to recruit healthy volunteers and collect blood samples for the above purpose. Each participant will be asked to donate 5 mL blood twice, at least one month apart.