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

miR-17~92 单倍体不足影响肺泡和血管内皮发育

• 批准号: 10310483
• 负责人: Mary E Robbins
• 金额: $ 16.31万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10310483
• 关键词: Advisory Committees; Affect; Alveolar; Alveolus; Animals; Architecture; Automobile Driving; Biological Markers; Biometry; Blood Vessels; Bronchopulmonary Dysplasia; Cells; Collagen; Communication; Cre lox recombination system; Development; Disease; Disease model; Dysplasia; Environment; Epigenetic Process; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Exhibits; Faculty; Foundations; Free Radicals; Functional disorder; Future; Gene Expression; Genes; Genetic; Genotype; Growth; Health; Heart Atrium; Human; Hyperoxia; Hypertension; Hypoxia; Immunohistochemistry; In Vitro; Infant; Injury; Junior Physician; Knock-out; Knowledge; Learning; Life; Link; Lung; Lung diseases; Matrix Metalloproteinases; Measurement; Medial; Mentors; Mentorship; MicroRNAs; Modeling; Mus; Neonatal; Neonatal Intensive Care; Outcome; Oxidation-Reduction; Oxygen; Pathogenesis; Pathogenicity; Patients; Pattern; Phase; Phenotype; Premature Infant; Production; Program Development; Proteins; Pulmonary Hypertension; Pulmonary artery structure; Pulmonary function tests; Pulmonary vessels; Recording of previous events; Reporting; Research Personnel; Sampling; Scientist; Signal Transduction; Signaling Protein; Supervision; System; Technical Expertise; Techniques; Testing; Thick; Tissue Sample; Transforming Growth Factor beta; Transgenic Model; United States; Vascular Endothelium; Viral Vector; Work; alveolar epithelium; angiogenesis; career; career development; cellular development; curative treatments; design; dosage; epigenetic silencing; experimental study; fibrotic lung; human tissue; improved; in vivo; in vivo Model; insight; lung development; lung injury; member; morphometry; mouse model; neonatal outcome; new therapeutic target; next generation sequencing; novel; novel diagnostics; overexpression; oxygen toxicity; postnatal; premature lungs; pressure; protein expression; pulmonary function; repaired; research and development; respiratory; restoration; skills; structural heart disease; targeted treatment; therapeutic target; transcriptome sequencing; whole genome

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.

项目总结 这份申请是为一个有指导的研究和职业发展计划，将使一个有动力的初级 医生-科学家发展必要的技能，最终成为一名独立的调查员。 该项目的成功完成将使调查员能够获得必要的基础和知识 开始研究导致支气管肺脏肺损伤的机制 不典型增生(BPD)，并可能开发靶向疗法来逆转或治愈这种损害。BPD是一种肺 每年影响14,000多名早产儿的疾病，对健康的长期影响 在患者的一生中影响肺功能。目前还没有针对BPD的根治疗法。在以前的工作中， 我们已经证明，一组被称为miR-17~92的microRNAs的表观遗传沉默与 在人类患者和该疾病的小鼠模型中出现严重的BPD。为了建立一种因果关系 在miR-17~92表达改变和BPD表型之间，该项目将研究中央 假设：小鼠miR-17~92簇的遗传部分缺失(或单倍体不足)将改变 肺泡和血管发育使人联想到BPD的表型，通过夸张的转化生长因子-β，胶原和 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单倍体功能不全对肺上皮细胞发育和功能的影响因果关系将会 通过使用病毒载体重新引入miR-17~92来建立功能恢复和 表型。由于有显著的细胞串扰驱动肺泡和血管内皮细胞的发育， 目的2将量化单倍性功能不全对肺血管内皮细胞发育和 功能。与目标1类似，将通过以下方式建立一个因果机制：返回函数实验 用病毒载体再次实现miR-17~92的全面表达。这项新颖的研究将通过以下方式影响该领域 了解miR-17~92被抑制和随后的修复所改变的细胞机制 集群。此外，这项研究具有重要的意义，因为它提供了一个研究细胞的模型 发生在传统BPD的肺泡和血管内皮细胞简化的机制 没有高氧暴露造成的令人困惑的自由基损伤的模型。上述概述 AIMS将在一个已有成功指导青少年的历史的环境中进行调查 教职员工走向独立。在专家咨询委员会的监督下，申请者将1) 她的技术技能，包括获得AS小鼠肺功能测试，免疫组织化学和NEXT 世代测序技术；2)学习高级生物统计学；3)学习 靶向治疗。未来的独立研究可能会集中在儿童与儿童之间的发育相互作用 肺泡和血管内皮细胞，以及miR-17~92簇作为生物标志物和/或治疗靶点的应用 对于BPD来说。

项目成果

Pathogenesis of Bronchopulmonary Dysplasia: Role of Oxidative Stress from 'Omics' Studies.

• DOI: 10.3390/antiox11122380
• 发表时间: 2022-12-01
• 期刊: Antioxidants (Basel, Switzerland)

Glutathione reductase deficiency alters lung development and hyperoxic responses in neonatal mice.

• DOI: 10.1016/j.redox.2020.101797
• 发表时间: 2021-01
• 期刊: Redox biology
• 影响因子: 11.4
• 作者: Robbins ME;Cho HY;Hansen JM;Luchsinger JR;Locy ML;Velten M;Kleeberger SR;Rogers LK;Tipple TE
• 通讯作者: Tipple TE