A State-of-Art NMR technique to Investigate Biologicals Effects of Electronic Nicotine Delivery Systems

采用最先进的 NMR 技术研究电子尼古丁输送系统的生物效应

基本信息

批准号：
10017237
负责人：
Jian Zhi Hu
金额：
$ 21.31万
依托单位：
BATTELLE PACIFIC NORTHWEST LABORATORIES
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-12 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10017237
关键词：
Acids Acrolein Address Aerosols Affect Airway Disease Aldehydes Amino Acids Biological Biomedical Research Cancer Patient Cell Extracts Cell Survival Cells Cigarette Coculture Techniques Detection Development Disease Progression Electronic Nicotine Delivery Systems Electronic cigarette Epithelial Epithelial Cells Epithelium Esters Event Flavoring Gases Gene Expression Gene Proteins Goals Health Health Benefit High temperature of physical object Human In Situ Individual Inflammation Inflammatory Investigation Knowledge Laboratories Lipids Liquid substance Lung Lung Inflammation Magic Malignant neoplasm of lung Mass Spectrum Analysis Measurement Metabolic Methods Minor Molecular Molecular Evolution Molecular Structure Monitor Nicotine Normal Cell Nuclear Magnetic Resonance Oxidation-Reduction Oxidative Stress Pathogenesis Poison Population Production Resolution Risk Factors Sampling Science Solid Solvents Stress Structure Structure of parenchyma of lung System Techniques Technology Temperature Time Tissue Extracts Tissues Tobacco Use Cessation base biological systems cancer cell cigarette smoking cytotoxicity electronic liquid electronic structure experimental group feature detection glycidol human disease indexing insight instrument metabolome metabolomics novel pressure quantum chemistry response time use tool vapor

项目摘要

PROJECT SUMMARY/ABSTRACT Cigarette smoking is a known risk factor for lung cancer, yet is also common among lung cancer patients. Electronic nicotine delivery systems (“ENDS”), i.e., “E-cigarettes”, are rapidly growing in popularity as a safer alternative to cigarettes to aid in tobacco cessation efforts, however, “safer” does not necessarily mean “safe”. There is a knowledge gap in our understanding of the molecular structures that comprise ENDS aerosols and the biological consequences of ENDS aerosols. This information is critically needed to develop an early understanding of the potential adverse health effects of ENDS in humans. To address this knowledge gap, the following two Specific Aims are proposed. Specific Aim 1: Investigation of the formation mechanism of ENDS aerosols at different temperatures, which will be enabled by using our recently developed in situ (a few kHz) magic angle spinning (MAS) technique that is capable of generating high resolution NMR spectra on samples containing a mixture of gases, liquids, and solids at significantly elevated temperature (from 0 to > 250C) and pressure (below 1 bar to >100 bars). This unique technique is ideal to determine how sensitive different E-liquid components are to aerosolization as a function of the temperature, including in particular the highly toxic chemicals such as aldehydes that can be generated by pyrolysis of ENDS solvents at high temperatures, and to determine the molecular structure of aerosols produced at different temperatures. Specific Aim 2: Application of non-destructive slow-MAS NMR metabolomics platform to define the dynamic response of lung organotypic cultures to ENDS aerosols. Slow-MAS NMR dramatically increases spectral resolution on intact biological tissues and cells, allowing detection of more metabolite features than can be resolved by conventional NMR instruments. The non-destructive capability of slow-MAS NMR (40-100Hz) is also uniquely suited for extended live cell/tissue measurements which is far superior to destructive approaches examining single time points. The feature of non-destructive detection is particularly advantageous as some metabolites only exist in live biological systems. We have developed a lung organotypic culture platform that enables us to investigate single cell populations (e.g. normal vs cancer cells), as well as mixed cell populations (e.g. normal/cancer cell co-cultures). We will use this system to define the baseline metabolome of normal human lung epithelial cells, lung cancer cells and their mixture as cocultures, as well as dynamic changes in these experimental groups induced by ENDS aerosols generated at different temperatures. Indices of toxic potential (cell viability, stress-responsive gene expression) will be defined under identical conditions. The goal is to determine if the production of ENDS at different temperatures alters the constituents and/or molecular structure of ENDS aerosols and their toxic potential on human cell systems by discovering and utilizing metabolic signatures.

项目概要/摘要吸烟是肺癌的已知危险因素，但在肺癌患者中也很常见。电子尼古丁输送系统（“ENDS”），即“电子烟”，作为一种更安全的电子烟，正在迅速普及。然而，“更安全”并不一定意味着“安全”。我们对组成 ENDS 气溶胶的分子结构的理解存在知识差距电子尼古丁传送系统气溶胶的生物学后果对于制定早期研究至关重要。了解电子尼古丁传送系统对人类潜在的不利健康影响。提出以下两个具体目标具体目标1：研究形成机制。在不同温度下ENDS气溶胶，这将通过使用我们最近开发的原位（一些 kHz）魔角旋转（MAS）技术，能够在含有气体、液体和固体混合物且温度显着升高（从 0 到 > 250°C）和压力（低于 1 bar 至 >100 bar），这种独特的技术非常适合确定灵敏度。不同的电子液体成分会随着温度的变化而雾化，特别包括 ENDS 溶剂在高温下热解可产生剧毒化学品，例如醛类温度，并确定不同温度下产生的气溶胶的分子结构。目标 2：应用非破坏性慢速 MAS NMR 代谢组学平台来定义动态肺器官培养物对 ENDS 气溶胶的响应显着增加了光谱。完整的生物组织和细胞的分辨率，允许检测更多的代谢特征慢速 MAS NMR (40-100Hz) 的无损能力也无法解决。独特地适合扩展活细胞/组织测量，远远优于破坏性方法检查单个时间点的无损检测的特点特别有利。代谢物仅存在于活体生物系统中。我们开发了一种肺器官培养平台。使我们能够研究单细胞群（例如正常细胞与癌细胞）以及混合细胞群（例如正常/癌细胞共培养）我们将使用该系统来定义正常人的基线代谢组。肺上皮细胞、肺癌细胞及其混合物作为共培养物，以及它们的动态变化不同温度下产生的 ENDS 气溶胶诱导的实验组的潜在毒性指数。（细胞活力、应激反应基因表达）将在相同条件下定义。确定不同温度下 ENDS 的生产是否会改变成分和/或分子结构通过发现和利用代谢，研究 ENDS 气溶胶及其对人体细胞系统的潜在毒性签名。