Human 3D Microtissues for Toxicity Testing via Integrated Imaging, Molecular and Functional Analyses

项目摘要 从历史上看，毒性试验依赖于动物的高剂量暴露， 外推到人群中的低水平暴露，（以高成本）产生很大的不确定性， 人类健康风险评估。布朗大学生物工程研究伙伴关系（BRP）-人类 通过集成成像、分子和功能分析进行毒性测试的3D微组织- 通过开发新的3D人体微组织作为桥接技术，实现了变革性的变化 基于整合成像、分子和功能分析， 环境化学品和新兴毒物的毒性测试。 该项目分为两个阶段。阶段1建立五种组织类型（前列腺，卵巢， 肺、脑、心脏），以解决这些预测生物学平台开发所面临的关键挑战： 再现性、生物复杂性、综合终点和人类变异性。第二阶段选择其中两个 具有足够剂量和时间响应的用于计算系统生物学分析的3D微组织模型 用于定义体外至体内外推和安全性评估的不利起点的数据。 与合作者和商业伙伴合作，BRP团队包括生物学， 工程、数学和医学，他们组成了预测生物学高级中心， （https：//www.brown.edu/research/projects/center-to-advance-predictive-biology/）。下一个工作 假设指导项目：体外病理学评估人类3D微组织中的一个 计算系统生物学框架确定毒物引起的安全性不利出发点 考核用于这些评估的高内容、高通量平台是3D微组织测试 系统，重新征服人类组织的分化特征和特征性细胞功能。 通过解决这些具体目标，将朝着改变毒性测试的目标取得进展： 具体目标1.利用工程解决方案创新3D微组织平台， 设计、共聚焦成像和高通量工作流程 第二章具体目标优化3D微组织作为预测生物学平台 第三章具体目标简化体外细胞的图像采集、重建和定量分析 三维微组织病理学评价 第四章具体目标在计算系统中集成成像、分子和功能端点 人类安全评估生物学框架 布朗大学的BRP将加速人类3D的开发和商业化 微组织平台作为动物毒性测试的替代品。

PROJECT SUMMARY Historically, toxicity testing has relied on high dose exposures in animals with default methods for extrapolating to low level exposures in human populations, producing (at high cost) great uncertainty for human health risk assessments. This Brown University Bioengineering Research Partnership (BRP)—Human 3D Microtissues for Toxicity Testing via Integrated Imaging, Molecular and Functional Analyses— implements a transformative change by developing novel 3D human microtissues as a bridging technology based on integrating imaging, molecular and functional analyses that will facilitate more rapid, cost-effective toxicity testing of environmental chemicals and emerging toxicants. The project has two phases. Phase 1 establishes 3D microtissues of five tissue types (prostate, ovary, lung, brain, heart) to address the key challenges facing development of these predictive biology platforms: reproducibility, biological complexity, integrated endpoints, and human variability. Phase 2 selects two of these 3D microtissue models for computational systems biology analysis with sufficient dose- and time-response data to define adverse points of departure for an in vitro-to-in vivo extrapolation and safety assessment. Working with collaborators and commercial partners, the BRP team includes faculty from biology, engineering, mathematics, and medicine who have formed the Center to Advance Predictive Biology (https://www.brown.edu/research/projects/center-to-advance-predictive-biology/). The following working hypothesis guides the project: In vitro pathology assessment of human 3D microtissues within a computational systems biology framework identifies toxicant-induced adverse points of departure for safety assessment. The high content, high throughput platforms for these evaluations are 3D microtissue test systems that re-capitulate the differentiated features and characteristic cellular functions of humans tissues. Progress toward the goal of transforming toxicity testing will be made by addressing these Specific Aims:  Specific Aim 1. Innovate the 3D microtissue platform with engineering solutions for improved well designs, confocal imaging, and high-throughput workflows  Specific Aim 2. Optimize 3D microtissues as predictive biology platforms  Specific Aim 3. Streamline image acquisition, reconstruction, and quantitative analysis for the in vitro pathology assessment of 3D microtissues  Specific Aim 4. Integrate imaging, molecular, and functional endpoints within a computational systems biology framework for the purpose of human safety assessment This Brown University BRP will accelerate development and commercialization of human 3D microtissue platforms as alternatives to animal toxicity testing.