Mesenchymal stromal cells and bronchopulmonary dysplasia

简介（由申请人提供）：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），一种早产儿慢性肺部疾病。在她的研究期间，Popova博士发表了4篇关于BPD和肺间充质间质细胞的论文。在拟议的培训期间，Popova博士将继续与她的导师Marc Hershenson一起工作，并接受联合导师Stephen Weiss（基质生物学专家）的额外科学指导。然而，她的主要目标将是获得所需的技能，成为一名独立的研究者，进行以患者为导向的研究，并将基础发现转化为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临床研究硕士课程，并接受费尔南多·马丁内斯博士的额外指导，他是一位成熟的肺病临床研究者。她还将在第三位共同导师Santiago Schnell博士的指导下学习计算医学和生物信息学课程。Popova博士的一般假设是间充质间质细胞肌成纤维细胞分化和基质细胞蛋白表达在BPD发病中起关键作用。为了验证这一点，她提出了三个具体目标，每个目标都与具体培训目标相匹配：测量肺间充质间质细胞的基因表达，重点是基质细胞蛋白。特异性假设：间充质间质细胞维持稳定的肌成纤维细胞分化表型，并与基质细胞蛋白表达相结合。早产儿气管吸入将检查基质细胞蛋白表达和间充质间质细胞。基质细胞mRNA和蛋白表达将通过微阵列和ELISA/免疫印迹检测。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蛋白表达与临床预后的关系特异性假设：基质细胞蛋白的表达预测BPD的发生。BPD作为气管吸入蛋白表达和间充质间质细胞基因表达的函数的相对几率将使用多变量回归分析来计算。培养目标：获得实验设计、统计学、计算医学和生物信息学方面的经验和理解。3. 检测早产儿肺部基质细胞蛋白的表达和定位。特异性假设：BPD患儿肺基质细胞蛋白表达增加。利用罗彻斯特大学新生儿肺生物库样本，将通过激光捕获显微镜、qPCR和基于立体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：获得用于定量人体组织小样本基因表达方法的经验/专业知识。这项工作的完成将有助于深入了解BPD的起源，并为Popova博士提供培训，促进她过渡到以患者为导向的研究的独立研究者。

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.