Delineating the Genetic Susceptibility of Smoking-Induced Vascular Dysfunction

项目总结 这份提案描述了一项为期五年的职业发展计划，为郭洪超博士的职业生涯做好准备 一名独立调查员。这个项目将建立在郭博士作为分子干细胞生物学家的背景之上 通过向他提供生物信息学、秘密组学和药物发现技术方面的专业知识，以及 环境健康、免疫学和心血管生物学，以促进我们对遗传 个体差异导致他们容易患上与吸烟有关的心血管疾病。郭博士会 由心血管专家、斯坦福大学心血管研究所所长约瑟夫·吴博士指导 使用诱导多能干细胞(IPSCs)建立疾病模型。此外，郭博士还将与郭广昌博士共同指导。 卡里·纳多，斯坦福大学肖恩·N·帕克过敏和哮喘研究中心主任， 世卫组织是研究包括烟草烟雾在内的气雾化污染物对免疫发展的影响的专家 原发免疫疾病、过敏和哮喘中的功能障碍。郭博士K99阶段的培训将包括 主要导师和共同导师的结构化指导，与合作者的密切互动， 与咨询委员会成员的补充会议，一个具有挑衅性的研究项目，以及一个量身定做的 职业发展和过渡计划。在郭博士之前的著作中，他概括了主要特征 使用患者特异性IPSC来源的内皮细胞(IPSC-1)研究尼古丁诱导的血管功能障碍 EC)。他还表明，烟碱型乙酰胆碱受体的基因变异加剧了尼古丁诱导的 通过增加炎症细胞因子的表达和细胞凋亡而导致的EC功能障碍，这使得对 IPSC-EC模型中的分子机制具有前所未有的深度和分辨率。与 高通量RNA测序及前沿分泌组学和药物筛选研究进展 技术，郭博士处于一个独特的位置，可以对患者尼古丁受体基因变异的风险进行分层 吸烟引起的血管疾病，并研究其可能的机制，最终目的是发现 心血管疾病和风险的生物标志物和精确治疗。在K99阶段，郭博士将产生 并表征了患者特定和等基因的IPSC-EC模型，用于研究三种不同的 尼古丁诱导的血管功能障碍的nAChR变体(目标1)。有了这个平台，郭博士将整合 高通量RNA测序和尖端分泌组学技术定义关键分子基础 和分泌体生物标记物的这些变异的风险吸烟介导性血管疾病(目标2)。在 R00阶段，郭博士将开发一个筛查平台，检查抗炎药物的有益效果 并在IPSC-EC中筛选携带这些风险变体的机制导向型小分子(目标3)。总而言之， 郭博士提出的工作将创建一个有价值的平台来评估这些变体与吸烟相关的风险 并揭示其分子机制和生物标志物。此外，这项工作将导致 机制导向治疗研究，将由郭博士作为独立研究员进行。

PROJECT SUMMARY This proposal describes a five-year career development program to prepare Dr. Hongchao Guo for a career as an independent investigator. This program will build on Dr. Guo’s background as a molecular stem cell biologist by providing him expertise in bioinformatics, secretomics, and drug discovery techniques, and knowledge on environmental health, immunology and cardiovascular biology to advance our understanding on how genetic differences in individuals contribute to their susceptibility to smoking-related cardiovascular disease. Dr. Guo will be mentored by Dr. Joseph Wu, Director of Stanford Cardiovascular Institute, who is an expert in cardiovascular disease modeling using induced pluripotent stem cells (iPSCs). In addition, Dr. Guo will be co-mentored by Dr. Kari Nadeau, Director of the Sean N. Parker Center for Allergy and Asthma Research at Stanford University, who is an expert in studying aerosolized pollutants including tobacco smoke on the development of immune dysfunction in primary immune disease, allergy and asthma. The K99 phase of Dr. Guo’s training will consist of structured mentorship by the primary mentor and co-mentor, close interactions with collaborators, complementary meeting with advisory committee members, a provocative research project, and a tailored program for career development and transition. In Dr. Guo’s previous works, he has recapitulated key features of nicotine-induced vascular dysfunction in patients using patient-specific iPSC-derived endothelial cell (iPSC- EC). He has also shown that genetic variants in nicotinic acetylcholine receptor exacerbated nicotine-induced EC dysfunction via increasing inflammation cytokines expression and apoptosis, which allow for analysis of the molecular mechanism in iPSC-EC model with a level of depth and resolution never before achieved. With the current advancement in high-throughput RNA sequencing and the cutting-edge secretomics and drug screening technologies, Dr. Guo is in a unique position to stratify the patient risk of genetic variants in nicotinic receptors for smoking-induced vascular diseases, and to study the potential mechanisms, with the ultimate goal to discover biomarkers and precise treatment for cardiovascular diseases and risks. In the K99 phase, Dr. Guo will generate and characterize patient-specific and isogenic iPSC-EC models for studying the susceptibility of three different nAChR variants to nicotine-induced vascular dysfunction (Aim 1). With the platform, Dr. Guo will then integrate high-throughput RNA sequencing and cutting-edge secretomics technologies to define the key molecular basis and secretomic biomarkers for the risk of these variants to smoking-mediated vascular diseases (Aim 2). In the R00 phase, Dr. Guo will develop a screening platform to examine the beneficial effect of anti-inflammatory drugs and screen mechanism-oriented small molecules in iPSC-EC carrying these risk variants (Aim 3). Collectively, Dr. Guo’s proposed work will create a valuable platform to evaluate the risk of these variants to smoking-related vascular disease and to reveal the molecular mechanisms and biomarkers. Additionally, this work will lead to studying of mechanism-oriented treatment, which will be carried out by Dr. Guo as an independent investigator.