Mesenchymal stromal cells and bronchopulmonary dysplasia

描述(申请人提供)：Antonia P.Popova，医学博士，儿科肺科医生，专注于支气管肺发育不良(BPD)，这是一种早产儿的慢性肺部疾病。在她担任研究员期间，Popova博士发表了4篇关于BPD和肺间充质基质细胞的论文。在拟议的培训期间，波波娃博士将继续与她的导师马克·赫申森合作，并接受共同导师、基质生物学专家斯蒂芬·韦斯的额外科学指导。然而，她的主要目标将是获得在以患者为导向的研究中成为独立调查者所需的技能，并将基本发现转化为BPD的临床问题。为了实现这一目标，Popova博士将报名参加公共卫生学院临床研究硕士项目，并接受费尔南多·马丁内斯博士的额外指导，费尔南多·马丁内斯博士是一名知名的肺部临床研究员。她还将在第三位共同导师圣地亚哥·施内尔博士的指导下学习计算医学和生物信息学方面的课程。Popova博士的一般假设是，间充质基质细胞、肌成纤维细胞分化和基质细胞蛋白表达在BPD的发病机制中起关键作用。为了验证这一点，她提出了三个特定的目标，每个目标都与特定的培训目标配对：1.测量肺间充质基质细胞的基因表达，重点是基质细胞蛋白。具体假设：间充质基质细胞维持稳定的肌成纤维细胞分化表型，并伴随着基质细胞蛋白的表达。将对早产儿的气管抽吸物进行基质蛋白表达和间充质基质细胞检查。采用基因芯片和酶联免疫印迹法检测基质细胞的基因和蛋白表达。培训目标：培养在实验设计、知情同意和研究团队管理方面的经验和理解。2.新生儿肺间充质基质细胞基质细胞基质蛋白表达与临床预后的关系具体假设：基质细胞蛋白的表达预测BPD的发生。BPD的相对优势作为气管吸液蛋白表达和间充质细胞基因表达的函数将使用多元回归分析来计算。培训目标：在实验设计、统计学、计算医学和生物信息学领域获得经验和理解。3.检测早产儿肺组织中基质细胞蛋白的表达和定位。特殊假设：BPD婴儿肺中基质细胞蛋白表达增加。使用罗切斯特大学新生儿肺生物库样本，将通过激光捕获显微镜、定量聚合酶链式反应和基于体视学的定量免疫组织化学来评估基质细胞蛋白的表达。培训目标：获得用于量化人类组织小样本中基因表达的方法的经验/专业知识。这项工作的完成将为BPD的起源提供洞察力，并为Popova博士提供培训，促进她在以患者为导向的研究中过渡为独立研究员。

DESCRIPTION (provided by applicant): Antonia P. Popova, M.D., a pediatric pulmonologist, is focusing her career on bronchopulmonary dysplasia (BPD), a chronic lung disease of prematurely-born infants. During her fellowship, Dr. Popova published 4 papers on BPD and lung mesenchymal stromal cells. During the proposed training, Dr. Popova will continue work with her Mentor, Marc Hershenson, and receive additional scientific direction from Co-Mentor Stephen Weiss, an expert in matrix biology. However, her main goal will to be to acquire skills needed to become an independent investigator in the conduct of patient-oriented research, and to translate basic discoveries to the clinical problem of BPD. To achieve this, Dr. Popova will enroll in the School of Public Health Clinical Research Masters Program and receive additional mentorship from Dr. Fernando Martinez, an established pulmonary clinical investigator. She will also take coursework in computational medicine and bioinformatics under the supervision of a third Co-Mentor, Dr. Santiago Schnell. Dr. Popova's general hypothesis is that mesenchymal stromal cell myofibroblastic differentiation and matricellular protein expression play critical roles in BPD pathogenesis. To test this, she proposes the three Specific Aims, each of which is paired with Specific Training Objectives: 1. Measure the gene expression of lung mesenchymal stromal cells, focusing on matricellular proteins. Specific hypothesis: Mesenchymal stromal cells maintain a stable phenotype of myofibroblastic differentiation that is coupled with matricellular protein expression. Tracheal aspirates of premature infants will be examined for matricellular protein expression and mesenchymal stromal cells. Stromal cell mRNA and protein expression will be measured by microarray and ELISA/immunoblotting. Training Objective: Develop experience and understanding in experimental design, informed consent, and management of a research team. 2. Correlate neonatal lung mesenchymal stromal cell matricellular protein expression with clinical outcomes. Specific hypothesis: Expression of the matricellular proteins predicts BPD development. Relative odds of BPD as a function of tracheal aspirate protein expression and mesenchymal stromal cell gene expression will be calculated using multivariate regression analysis. Training Objectives: Acquire experience and understanding in the areas of experimental design, statistics, computational medicine and bioinformatics. 3. Examine expression and localization of matricellular proteins in the lungs of premature infants. Specific hypothesis: Matricellular protein expression is increased in the lungs of infants with BPD. Using University of Rochester neonatal lung biorepository samples, matricellular protein expression will be assessed by laser capture microscopy, qPCR and stereology-based quantitative immunohistochemisty. Training Objective: Acquire experience/expertise in methods used to quantify gene expression in small samples of human tissue. Completion of this work will provide insight into the origins of BPD and provide training for Dr. Popova, facilitating her transition to independent investigator in patient-oriented research.