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Development of a High-Throughput Platform for Screening Ovotoxic Chemicals (K01ES030014)

开发用于筛选卵毒性化学品的高通量平台（K01ES030014）

基本信息

批准号：
10269942
负责人：
Shuo Xiao
金额：
$ 16.24万
依托单位：
RUTGERS, THE STATE UNIV OF N.J.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-25 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10269942
关键词：
3-Dimensional Alginates Animals Apoptosis Architecture Cell Death Cells Chemical Exposure Chemicals Competence Consumption Cryopreservation Dependence Development Dose Encapsulated Ensure Environmental Impact Environmental Pollution Evaluation Female Fertilization Germ Cells Goals Gold Gonadal structure Growth Health Hormone imbalance Hormone secretion Hormones Human Hydrogels Image In Vitro Individual Industrialization Infertility Laboratory Animals Measures Meiosis Methods Mitochondria Molecular Monitor Mus Oocytes Organelles Ovarian Ovarian Follicle Ovary Ovulation Poison Pregnancy Preventive Rapid screening Reaction Time Risk Somatotropin Stains System Technology Temperature Testing Time base cell injury confocal imaging cost cryogenics environmental chemical experimental study female reproductive system high throughput screening imaging system in vitro Model in vivo Model movie oocyte maturation ovarian failure ovotoxicity reproductive function reproductive organ reproductive outcome screening serial imaging steroid hormone

项目摘要

PROJECT SUMMARY Increasing evidence shows that a broad spectrum of both industrial and environmental chemicals can cause female ovarian toxicity (ovotoxicity) and increase the risk of hormonal imbalance, ovarian failure, and infertility in both humans and other mammalian species. The ovary is the primary female reproductive organ and functions to synthesize and secret steroid hormones and to support the oocyte maturation for ovulation, fertilization, and pregnancy. Ovotoxicity can be measured in two ways, from the health of the developing ovarian follicle and gamete and from the ability of the gonads to produce hormones. Unfortunately, the lack of optimal in vitro models makes the current gold standard for testing the ovotoxicity of chemicals rely on the use of laboratory animals. However, in vivo models are time-consuming, costly, and harmful to animals. In our previous studies, we have used an alginate hydrogel-based encapsulation method to grow both mouse and human preantral follicles outside of the context of the ovary, which is termed encapsulated in vitro follicle growth (eIVFG). The eIVFG maintains the 3D architecture of the follicles and supports follicle growth, hormone secretion as well as oocyte maturation and ovulation, suggesting a promising model for in vitro ovotoxicity testing. However, this method is relatively slow, and multiple rounds of follicle isolation, culture, and chemical exposure are necessary to complete dose-response and time-course experiments, making the current eIVFG scenario not optimal, particularly when a rapid screening is required in the case of unpredictable and emerging environmental threats. In this proposed project, we hypothesize that we can use cryopreservation technology and a confocal high-content imaging system to develop a high-throughput platform for screening ovotoxic chemicals. In Aim 1, we will bank the isolated ovarian follicles through vitrification for establishing a long-term and ready-to-use follicle storage platform. In Aim 2, we will use the ImageXpress confocal high-content imaging system to develop a high-throughput platform for follicle monitoring, assessing, and analysis. We expect that these proposed studies will allow us to obtain rapid and reliable evidences to understand the impacts of environmental contaminants on the female reproductive system, to provide guidance to conduct preventive and protective activities, as well as to minimize the dependence and use of animals.

项目摘要越来越多的证据表明，工业和环境化学品的广泛使用会导致女性卵巢毒性（ovotoxicity），并增加激素失衡，卵巢衰竭和不孕症的风险在人类和其他哺乳动物物种中。卵巢是女性的主要生殖器官，合成和分泌类固醇激素，并支持卵母细胞成熟排卵，受精，怀孕卵巢毒性可以通过两种方式来测量，从发育中的卵泡的健康状况，配子和性腺产生激素的能力。不幸的是，缺乏最佳的体外模型，使得目前测试化学品卵毒性的金标准依赖于实验室动物的使用。然而，体内模型耗时、成本高且对动物有害。在我们以前的研究中，我们有使用基于藻酸盐水凝胶的包封方法培养小鼠和人类腔前卵泡在卵巢环境之外，这被称为体外包裹卵泡生长（eIVFG）。关于eIVFG 维持卵泡的3D结构，支持卵泡生长、激素分泌以及卵母细胞成熟和排卵，这表明一个有前途的模型，在体外卵毒性测试。但是此方法相对较慢，需要多轮卵泡分离、培养和化学暴露才能完成剂量-反应和时间-过程实验，使目前的eIVFG方案不是最佳的，特别是当在出现不可预测和新出现的环境威胁时，需要进行快速筛查。在此提出的项目，我们假设我们可以使用冷冻保存技术和共聚焦高分子量成像，系统开发一个高通量筛选卵毒性化学品的平台。在目标1中，我们将玻璃化冷冻法分离卵巢卵泡，建立一个可长期保存、可移植的卵泡保存平台。在目标2中，我们将使用ImageXpress共聚焦高荧光含量成像系统来开发高通量的荧光成像系统。卵泡监测、评估和分析平台。我们希望这些拟议的研究将使我们能够获得快速可靠的证据，了解环境污染物对女性的影响生殖系统，提供指导，开展预防和保护活动，以及尽量减少对动物的依赖和利用。