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

项目总结 卵巢及其功能单位，即卵泡，是分泌性类固醇和多肽激素的关键。 以及卵母细胞的成熟和释放，以供排卵和受精。越来越多的证据表明 广泛的环境化学品和药物导致卵巢毒性(卵毒性)和 增加生育年龄妇女卵巢早衰、激素失衡和不孕症的风险 女人和青春期前的女孩。有超过85,000种化学品用于消费产品以及新兴市场 污染物，如有害的藻华(HAB)毒素，几乎没有方法来筛选潜在的毒性 卵巢。我们之前开发了一种芯片上卵巢(OvaryChip)，它保持了3D架构 并产生人类月经周期样的卵泡发育、激素分泌、卵母细胞成熟、 排卵和黄体生成。然而，这种创新模式的一个剩余挑战是，迫切需要 提高吞吐量。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……里面 目的1、将卵泡玻璃化冷冻、三维体外卵泡生长和一种新型的高含量卵泡芯片 培养和成像，以开发一种高通量的生长卵泡的卵毒性测试。在目标2中，我们将开发 一种利用三重转基因小鼠卵巢外植体检测原始卵泡卵子毒性的新芯片。 在目标3中，我们将开发一种芯片上的肝脏-卵巢，将肝脏代谢纳入卵子毒性测试。在这些 研究中，我们将从形态、功能和分子方面评估卵泡和卵母细胞的生殖结果。 1)具有已知卵毒性的化学品(验证研究)和2)单独和混合使用的新型HAB毒素的水平。 这些研究具有至关重要的意义，因为OvaryChip模型将(1)将一种单一的 将化合物转化为一个强大的高通量卵毒性筛选平台；(2)揭示对化学物质- 诱导的卵子毒性(阶段和代谢物依赖的机制)；(3)最大限度地减少成本和使用 动物；以及(4)优先考虑高卵毒关注的化学品，包括赤潮毒素，以进行进一步的风险评估。

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.