An ovary-on-a-chip to identify ovarian toxicity

项目摘要 卵巢及其功能单位卵泡对性类固醇和肽类激素的分泌至关重要 以及用于排卵和受精的卵母细胞的成熟和释放。越来越多的证据表明 广泛的环境化学品和药物引起卵巢毒性（卵毒性）， 增加卵巢早衰、激素失衡和育龄妇女不孕的风险 女性和青春期前的女孩超过85，000种化学品用于消费品以及新兴的 污染物，如有害藻华（HAB）毒素，有一些手段来筛选潜在的毒性， 卵巢我们之前开发了一种保持3D架构的芯片上卵巢（OvaryChip） 并产生人类月经周期样卵泡发育，激素分泌，卵母细胞成熟， 排卵和黄体化。然而，这一创新模式仍然面临的一个挑战是， 加快吞吐量。这项研究的中心假设是，在体外微流控卵泡， 培养物可用于鉴定环境毒物对卵巢毒性的新机制。在 目的1、将卵泡玻璃化冷冻、三维体外卵泡生长技术和高含量的卵巢芯片技术相结合 培养和成像以开发生长卵泡的高通量卵毒性测试。在目标2中，我们将开发 一种新的卵巢芯片用于三重转基因小鼠卵巢组织块的原始卵泡卵毒性检测。 在目标3中，我们将开发一种肝脏-卵巢-芯片，将肝脏代谢纳入卵毒性试验。在这些 研究，我们将评估卵泡和卵母细胞的生殖结果在形态，功能和分子， 1）已知具有卵毒性的化学品（验证研究）和2）单独和混合的新型HAB毒素。 这些研究是至关重要的，因为OvaryChip模型将（1）将工作台测定转化为单个 化合物到一个强大的高通量卵毒性筛选平台;（2）揭示新的见解化学- 诱导的卵毒性（阶段和代谢依赖机制）;（3）最大限度地减少成本和使用整个 （4）优先考虑具有高度卵毒性的化学品，包括有害藻华毒素，以作进一步的风险评估。

PROJECT SUMMARY The ovary and its functional unit, the follicles, are critical for the secretion of sex steroids and peptide hormones as well as the maturation and release of oocytes for ovulation and fertilization. Increasing evidence has revealed that a wide spectrum of environmental chemicals and pharmaceuticals cause ovarian toxicity (ovotoxicity) and heighten the risk of premature ovarian failure, hormonal imbalance, and infertility in both reproductive aged women and prepubertal girls. With over 85,000 chemicals used in consumer products as well as emerging contaminants such as harmful algal bloom (HAB) toxins, there are few means to screen for potential toxicity to the ovaries. We have previously developed an ovary-on-a-chip (OvaryChip) that maintains the 3D architecture of follicles and produces human menstrual cycle-like follicle development, hormone secretion, oocyte maturation, ovulation, and luteinization. Yet, one remaining challenge of this innovative model is the critical need to accelerate throughput. The central hypothesis of this research is that in vitro microfluidic follicular cultures can be used to identify novel mechanisms of ovarian toxicity for environmental toxicants. In Aim 1, we will integrate follicle vitrification, 3D in vitro follicle growth, and a new OvaryChip with high-content culture and imaging to develop a high-throughput ovotoxicity testing of growing follicles. In Aim 2, we will develop a new OvaryChip for ovotoxicity testing of primordial follicles by using triple transgenic mouse ovarian explants. In Aim 3, we will develop a liver-ovary-on-a-chip to incorporate liver metabolism into ovotoxicity testing. In these studies, we will assess follicle and oocyte reproductive outcomes at morphological, functional, and molecular levels for 1) chemicals with known ovotoxicities (validation studies) and 2) novel HAB toxins alone and in mixtures. These studies are critically significant because the OvaryChip models will (1) translate a bench assay for a single compound into a robust high-throughput ovotoxicity screening platform; (2) reveal novel insight into chemical- induced ovotoxicity (stage- and metabolite-dependent mechanisms); (3) minimize the cost and use of whole animals; and (4) prioritize chemicals of high ovotoxicity concern, including HAB toxins, for further risk assessment.