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Endothelial Progenitor Cells for Assessing Toxicological Response Variability in Humans

用于评估人类毒理学反应变异性的内皮祖细胞

基本信息

批准号：
9019006
负责人：
John William Ludlow
金额：
$ 47.43万
依托单位：
BRITEBIO, INC.
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9019006
关键词：
Acids Adult Affect Algorithms Animal Model Animals Automation Biological Assay Blood Blood Circulation Blood Vessels Blood specimen CD34 gene Cadmium Cell Culture Techniques Cell Cycle Arrest Cell Death Cell Differentiation process Cell Line Cell Separation Cell Survival Cells Chemicals Communities Contracts Culture Media Data Data Set Detection Development Developmental Process Differentiation Antigens Donor Selection Dose Endothelial Cells Environmental Health Evaluation Exhibits Exposure to Flow Cytometry Future Glycocalyx Goals Human Image In Vitro Individual Ionizing radiation Link Liquid substance Maintenance Mediating Methods Modeling Oxidative Stress PECAM1 gene Pathway interactions Phase Physiology Population Procedures Production Proteins Reactive Oxygen Species Reagent Reporting Research Risk Assessment Scientist Services Signal Transduction Source Specific qualifier value Stem cells Time Toxic Environmental Substances Toxic effect Toxicant exposure Toxicology Toxin Translating Umbilical cord structure Validation Western Blotting base bisphenol A cadherin 5 cell growth cell type commercialization electric impedance environmental agent experience high throughput screening imaging platform immortalized cell in vivo in vivo Model insight novel peripheral blood phase 1 study programs public health relevance repository response screening toxicant toxicant screening von Willebrand Factor

项目摘要

DESCRIPTION (provided by applicant): Toxicological risk assessment lacks direct estimation of the magnitude and variability of human responses to environmental toxicants. Specifically, there is very little data on the effects of low dose toxicants on human developmental processes, including cellular differentiation. That is because the current basis of toxicological assessment, animals and immortalized cell lines, cannot adequately model the impact of low level toxicant exposure on human populations. An alternative detection platform should enable incorporation of human variability in screening of a large number of toxicants in a high-throughput manner. To establish such a platform, we propose using human donor-specific cells that can be (a) isolated in a non-invasive manner, (b) easily expanded in culture, and (c) cryopreserved without loss of viability. Our preliminary data suggest that human endothelial progenitor cells, or endothelial colony forming cells (ECFCs), a population of CD31+/CD34+ pluripotent cells found in circulating blood, would fulfill these requirements. ECFCs are highly proliferative in vitro and ca differentiate into mature endothelial cells (ECs). The overall goal of this research is to evaluate the effect of chemical toxicants on viability, proliferation, and differentiation of ECFCs derived from different individuals. Prior to our recent and successful completion of a Phase I program, we established cell culture parameters for ECFCs using a proprietary cell growth media supplement and completed initial toxicological risk assessment studies using low-dose ionizing radiation (LDIR). These studies confirmed the utility of ECFCs in toxicological risk assessment screening as well as their ability to broadly represent varying responses expected in human populations. Phase I studies successfully expanded these findings by performing ECFC toxicology assays using Bisphenol A (BPA), perfluorooctanoic acid (PFOA) and cadmium (Cd). ECFCs were found to be sensitive to BPA and Cd and less sensitive to PFOA, exhibiting donor-specific variability in proliferation rates. BPA and Cd also induced reactive oxygen species (ROS) production in a donor-specific manner. ECFCs were shown to experience oxidative stress and cell cycle arrest after exposure to toxicants. Elevated ROS production was shown to correlate with cell death after longer incubation times with toxicants. We found that the toxicants we analyzed also affected expression of endothelial-specific cell markers at a protein level in a newly developed differentiation assay. Phase II will focus on the continued development of this screening platform toward a validated quantitative high content imaging-based screen. Commercialization of the platform as a service will also be pursued as well as the production of kits for ECFC culture and subsequent toxicological analysis. Finally, the screen will be expanded to include toxicant-induced effects on ECFC differentiation into mature endothelial cells.