Establishment of human mini-Testis for reproductive toxicity testing

用于生殖毒性测试的人体迷你睾丸的建立

基本信息

批准号：
10010729
负责人：
Lei Yin
金额：
$ 22.5万
依托单位：
REPROTOX BIOTECH, LLC
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：
2020-08-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10010729
关键词：
3-Dimensional Animal Testing Animals Biological Assay Biological Models Cardiovascular system Cell Cycle Cell Differentiation process Cells Chemicals Classification Coculture Techniques Cytoskeleton DNA Damage Data Development Dose Drug or Chemical Environment Exposure to Extracellular Matrix Germ Cells Grant Growth Health Human In Vitro Industrialization Knowledge Malignant Neoplasms Methods Mission Modeling Modernization Molecular Morphology National Research Council Occupational Exposure Oxidative Stress Pathway interactions Prevention Procedures Process Publications Reporting Reproducibility of Results Research Risk Risk Assessment Rodent Rodent Model Sensitivity and Specificity Spermatogenesis Spermatogonia Steroid biosynthesis System Testing Testis Time Toxic effect Toxicity Tests Toxicology Translational Research Vision Workplace adverse outcome base cost cost efficient developmental toxicity human embryonic stem cell human model in vitro Model in vivo information model leydig interstitial cell male predictive modeling programs reproductive reproductive toxicity research to practice screening sertoli cell stem cell differentiation stem cells toxicant

项目摘要

Summary Occupational exposures during manufacture and application of these chemicals represent the highest level of exposure that could cause potential adverse health effects. There remains a considerable knowledge gap regarding the relationship between workplace exposure of industrial chemicals and possible adverse health effects. Current test methods set by regulatory agencies predominately rely on animal testing. The cost and time with the animal testing for risk assessment seriously limit efforts for extensive chemical screening. These data gaps necessitate the implementation of a new vision for toxicity testing as reflected in the National Research Council’s report “Toxicity Testing in the 21st Century”. Under the support of NIOSH R21 OH010473 grant, we established a rodent testicular cell co-culture model and found this in vitro co-culture model could identify the reproductive toxicants. While results from these in vivo or in vitro rodent models are informative, these models could not recapitulate the whole process of human spermatogenesis as rodent spermatogenesis proceeds in a distinctly different fashion compared to humans. Thus, developing in vitro human models for reproductive toxicity testing is paramount for advancing the field and for understanding the cellular and molecular mechanisms that may underlie these disruptions in human spermatogenesis. In this proposal, we will develop a human testicular cell co-culture model, and demonstrate the reliability and sensitivity of this model in high-throughput and multi-parametric high-content analytical screening for workplace chemicals on the reproductive and developmental systems. The specific aims are (1) to establish testicular cell co-culture model from human spermatogonial stem cells (hSSCs), Sertoli cell and Leydig cells; (2) to develop integrated assays based on the adverse outcome pathways; (3) to apply the multi-parametric high-throughput and high- content analysis (HT/HCA) to examine and compare reproductive toxic and non-toxic compounds to validate the in vitro testicular cell co-culture model. Through this hypothesis-driven research, we will test the hypothesis that an in vitro 3D human testicular cell co-culture model creates a proper microenvironment that mimics a human testis to promote the growth of germ cells in vitro and generate reproducible results for predicting reproductive toxicity. The research proposed will result in publications and generate toxicity data of the chemicals in the workplace, and provide data for risk assessment. Therefore, this proposal meets the mission of Cancer, Reproductive, and Cardiovascular Research Program (CRC). The identification of reproductive toxicants will reduce hazardous exposures and lower the risk of reproductive malfunction. Therefore, our proposed project is within the NIOSH CRC’s Research to Practice (r2p) strategy to promote the transfer and translation of research findings into prevention practices.

概括这些化学物质的制造和应用期间的职业暴露是最高的可能导致潜在不良健康影响的暴露水平。仍然存在考虑知识差距关注工业化学物质的工作场所暴露与可能的不良健康之间的关系效果。监管机构设定的当前测试方法主要依赖于动物测试。成本和随着动物测试进行风险评估的时间，严重限制了广泛的化学筛查工作。这些数据差距必须实施对毒性测试的新愿景，如国家研究委员会的报告“ 21世纪的毒性测试”。在NIOSH R21 OH010473的支持下格兰特，我们建立了一个啮齿动物验证细胞共培养模型，发现这种体外共培养模型可以确定生殖有毒物质。虽然这些体内或体外啮齿动物模型的结果是有益的，但这些模型无法概括人类精子发生的整个过程与人类相比，以明显不同的方式进行收益。那是为了开发体外人类模型生殖毒性测试对于前进和了解细胞和细胞和可能是人类精子发生中这些破坏的分子机制。在这个建议中，我们将开发人类的测试细胞共培养模型，并证明这一点的可靠性和敏感性在高通量和多参数高容量分析筛查工作场所化学品上的模型具体目的是（1）建立验证细胞共培养来自人类精子干细胞（HSSC），Sertoli细胞和Leydig细胞的模型；（2）发展综合基于不良结果途径的测定；（3）应用多参数高通量和高级内容分析（HT/HCA）检查和比较复制性有毒和无毒化合物以验证体外验证细胞共培养模型。通过这项假设驱动的研究，我们将测试假设体外3D人类测试细胞共培养模型创造了适当的微环境模仿人睾丸以促进体外生长的生长，并产生可重复的结果预测生殖毒性。提出的研究将导致出版物并产生毒性数据工作场所中的化学品，并提供风险评估的数据。因此，该建议符合癌症，生殖和心血管研究计划（CRC）的任务。识别生殖有毒物质将减少危险暴露并降低生殖风险故障。因此，我们提出的项目在NIOSH CRC的研究中（R2P）。促进将研究结果转移和翻译为预防实践的策略。