Delineating the Genetic Susceptibility of Smoking-Induced Vascular Dysfunction

项目摘要 本计划书描述了一个为期五年的职业发展计划，旨在为郭洪超博士的职业生涯做好准备， 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 通过为他提供生物信息学、分泌组学和药物发现技术方面的专业知识， 环境健康，免疫学和心血管生物学，以促进我们对遗传如何 个人的差异导致他们对吸烟相关的心血管疾病的易感性。郭医生会 由斯坦福大学心血管研究所所长Joseph Wu博士指导，他是心血管领域的专家 使用诱导多能干细胞（iPSC）的疾病建模。此外，郭博士将由郭博士共同指导。 Kari Nadeau，Sean N.斯坦福大学帕克过敏和哮喘研究中心， 他是研究烟雾污染物（包括烟草烟雾）对免疫系统发展的专家。 在原发性免疫疾病、过敏和哮喘中的功能障碍。郭博士的K99培训阶段将包括 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，一个挑衅性的研究项目，以及一个量身定制的 职业发展和转型计划。在郭博士以前的作品中，他概括了 尼古丁诱导的血管功能障碍的患者使用患者特异性iPSC衍生的内皮细胞（iPSC- EC）。他还表明，尼古丁乙酰胆碱受体的遗传变异加剧了尼古丁诱导的 通过增加炎症细胞因子表达和细胞凋亡导致EC功能障碍，这使得可以分析EC功能障碍的原因。 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测序的最新进展以及最前沿的分泌组学和药物筛选 技术，郭博士是在一个独特的位置，以分层患者的风险，遗传变异的烟碱受体 吸烟引起的血管疾病，并研究潜在的机制，最终目标是发现 生物标志物和精确治疗心血管疾病和风险。在K99阶段，郭博士将产生 并表征患者特异性和同基因iPSC-EC模型，用于研究三种不同的iPSC-EC易感性。 nAChR变体对尼古丁诱导的血管功能障碍的影响（目的1）。有了这个平台，郭博士接下来将整合 高通量RNA测序和尖端的分泌组学技术，以确定关键的分子基础 以及这些变异体对吸烟介导的血管疾病的风险的分泌组生物标志物（目的2）。在 R 00阶段，郭博士将开发一个筛选平台，以检查抗炎药物的有益效果 并在携带这些风险变体的iPSC-EC中筛选机制导向的小分子（Aim 3）。总的来说， 博士郭的提议工作将创建一个有价值的平台，以评估这些变体对吸烟相关的风险。 血管疾病，并揭示分子机制和生物标志物。此外，这项工作将导致 机制导向治疗的研究，将由郭博士作为独立研究者进行。

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.