PCOS and androgen-related disease modeling and drug testing in Multi-organ Integrated Microfluidic Reproductive Platform

项目摘要 已经开发出支持包括卵泡在内的28天生殖周期的微流控系统 生长、排卵和黄体化，伴随着雌二醇和黄体酮的变化。这个 在这种情况下从卵泡中释放出来的卵母细胞是健康的，并具有预期的核和 体内排卵卵母细胞的胞质成熟表型。我们已经整合了卵巢组织和 将激素谱循环到包括人输卵管组织的多路微动力学单元中， 子宫内膜、宫颈和肝脏的细胞器。我们还分别改造了小鼠和人类的胰岛 睾丸和前列腺进入类似的生物活性系统。此应用程序的目的是通过以下方式进一步推进我们的工作 实施下一代微流控系统，该系统是为高 吞吐量机器人设置，将使药物测试集成器官系统模拟各种 生殖疾病。将要检验的假设是，我们可以创建一个体外微流控系统 这代表了多囊卵巢综合征的特征，这是一种多器官疾病，影响8%-10%的 育龄妇女和谁没有合适的体外模型。在过去四年的工作中， 我们开发了第一代微流控平台，允许激素和组织水平的功能 上面。在本申请的目标1中，我们将最终确定专门为机器人技术构建的新系统 实验室。该系统由不吸收类固醇的材料制成，并具有泵送配置文件 稳定可达一个月。原型也已经被机器人处理了。板载控制器、泵和 电池已经被改进，目标是一种低成本、可重复使用的设备，可以轻松地在 孵化器以及较大规模的筛选实验室。我们的目标是开发一种设备，它将取代所有 普通的平板培养工作使生物学家可以不再研究静态废物中的扁平细胞-- 累积模型。在目标2中，我们将为每个器官和相连的器官建立多囊卵巢综合征模型 并在Aim 3中测试各种改变雄激素或胰岛素代谢的药物。这将是 与阿斯利康的合作结束了。综上所述，我们的研究将提供一个重要的下一代工具箱 妇女的健康和更广泛的细胞生物学领域。

Project Summary Microfluidic systems have been developed that support 28 day reproductive cycles including ovarian follicle growth, ovulation, and luteinization with the accompanying changes in estradiol and progesterone. The oocytes that are released from the follicle in this setting are healthy and have the predicted nuclear and cytoplasmic maturation phenotypes of in vivo ovulated oocytes. We have integrated the ovarian tissue and cycling hormone profiles into a multiplexed microdynamic unit that includes human fallopian tube tissue, uterine endometrium, cervix, and liver organoids. We have also adapted mouse islets and separately, human testis and prostate into similarly bioactive systems. The purpose of this application is to further our work by implementing a next generation microfluidic system that has been created for the express purpose of a high throughput robotics setting that will enable drug testing of integrated organ systems that mimic a variety of reproductive diseases. The hypothesis that will be tested in that we can create an in vitro microfluidic system that represents hallmarks of polycystic ovary syndrome, a multiorgan disease that affects 8-10% of reproductive age women and for whom there is no adequate in vitro model. During the past 4 years of work, we developed the first generation microfluidic platform that permitted the hormone and tissue level function above. In aim 1 of the present application, we will finalize a new system that was built expressly for a robotics laboratory. This system is made of materials that does not absorb steroids and has pumping profiles that are stable up to one month. Prototypes have also been handled robotically. Onboard controllers, pumps and batteries have been adapted with the goal of a low cost, reusable device that could be easily used in an incubator as well as the larger scale-screening laboratory. Our goal is to develop a device that will replace all ordinary plated culture work so that biologists can move away from studying flat cells in static waste- accumulating models. In aim 2 we will establish models of PCOS for each organ and for the organs connected to each other and test a variety of drugs in aim 3 that will alter androgen or insulin metabolism. This will be done with Astrazeneca. Taken together, our studies will provide a next generation toolbox important to women's health and to the broader field of cell biology.