miR-17~92 Haploinsufficiency Influences Alveolar and Vascular Endothelial Development

项目概要 该申请适用于指导研究和职业发展计划，该计划将使有动力的初级学生 医师科学家培养最终成为独立研究者所需的技能。 成功完成该项目将使研究者能够获得必要的基础和知识 开始研究支气管肺损伤的机制 发育异常（BPD）并有可能开发出靶向疗法来逆转或治愈这种损害。 BPD是肺 该疾病每年影响超过 14,000 名早产儿，并对健康造成长期影响 影响患者一生的肺功能。目前尚无针对 BPD 的治疗方法。在之前的工作中， 我们已经证明，一簇 microRNA（称为 miR-17~92）的表观遗传沉默与 人类患者和该疾病的小鼠模型中出现严重的 BPD。为了建立因果关系 miR-17~92 表达改变与 BPD 表型之间的关系，本项目将研究其中枢 假设： miR-17~92 簇的小鼠遗传部分缺失（或单倍体不足）会改变 肺泡和血管发育通过夸大的 TGF-β、胶原蛋白和 基质金属蛋白酶的表达。在目标 1 中，将使用体内和体外系统来确定效果 miR-17~92单倍体不足对肺上皮发育和功能的影响。因果关系将 通过使用病毒载体重新引入 miR-17~92 来确定功能的恢复和 表型。由于存在显着的细胞串扰驱动肺泡和血管内皮发育， 目标 2 将量化单倍体不足对肺血管内皮发育和 功能。与目标 1 类似，通过回归功能实验将建立因果机制 通过病毒载体重新引入miR-17~92的完整表达。这项新颖的研究将通过以下方式影响该领域 了解 miR-17~92 的抑制和随后恢复所改变的细胞机制 集群。此外，这项研究很重要，因为它提供了一个研究细胞的模型 导致传统 BPD 中肺泡和血管内皮细胞简化的机制 模型没有因高氧暴露引起的混淆自由基损伤。以上概述的 将在具有成功指导初级学生的既定历史的环境中对目标进行调查 教师的独立性。在专家咨询委员会的监督下，申请人将 1) 推进 她的技术技能，包括小鼠肺功能测试、免疫组织化学等 世代测序技术； 2）学习先进的生物统计学； 3）学习交付机制 靶向治疗。未来的独立研究可能会集中于两者之间的发展相互作用 肺泡和血管内皮，以及使用 miR-17~92 簇作为生物标志物和/或治疗靶点 对于边缘型人格障碍。

PROJECT SUMMARY This application is for a mentored research and career development program that will enable a motived junior physician-scientist to develop the skills necessary to ultimately become an independent investigator. Successful completion of this project will enable the investigator to gain the sills and knowledge necessary to launch a career investigating the mechanisms responsible for the lung damage present in bronchpulmonary dysplasia (BPD) and potentially develop targeted therapeutics to reverse or cure this damage. BPD is a lung disease that affects more than 14,000 premature infants each year with prolonged health ramifications that affect lung function throughout the patient's life. No curative therapies currently exist for BPD. In previous work, we have shown that epigenetic silencing of a cluster of microRNAs, called miR-17~92, is associated with severe BPD in human patients and murine models of the disease. In order to establish a causal relationship between altered miR-17~92 expression and a BPD phenotype, this project will investigate the central hypothesis that: murine genetic partial-deletion (or haploinsufficiency) of the miR-17~92 cluster will alter alveolar and vascular development reminiscent of a BPD phenotype via exaggerated TGF-β, collagen and matrix metalloproteinase expression. In Aim 1, in vivo and in vitro systems will be used to determine the effects of miR-17~92 haploinsufficiency on pulmonary epithelial development and function. A causal relationship will be determined by reintroduction of miR-17~92 using a viral vector to establish return of function and phenotype. Since there is significant cellular cross-talk driving alveolar and vascular endothelial development, Aim 2 will quantify the effects of haploinsufficiency on pulmonary vascular endothelial development and function. Similar to Aim 1, a causal mechanism will be established with return of function experiments by reintroduction of the full expression of miR-17~92 by viral vector. This novel study will impact the field through understanding the cellular mechanisms altered by suppression and subsequent restoration of the miR-17~92 cluster. Furthermore, this study is significant because it provides a model to investigate the cellular mechanisms responsible for alveolar and vascular endothelial simplification that occurs in traditional BPD models without the confounding free-radical damage caused by high oxygen exposure. The above outlined aims will be investigated in an environment with an established history of successful mentorship of junior faculty to independence. Under the supervision of an expert advisory committee, the applicant will 1) Advance her technical skills, with acquisition of as murine pulmonary function testing, immunohistochemistry, and next generation sequencing techniques; 2) Learn advanced biostatistics; and 3) Learn delivery mechanisms of targeted therapeutics. Future independent studies will likely focus on the developmental interplay between the alveoli and vascular endothelium, and use of the miR-17~92 cluster as a biomarker and/or therapeutic target for BPD.