Pulmonary Endothelial-Mesenchymal Crosstalk in Hyperactive mTOR Lung Diseases

mTOR 肺疾病中的肺内皮-间质串扰

基本信息

批准号：
10388900
负责人：
Susan M Lin
金额：
$ 2.5万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-01-01 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10388900
关键词：
Affect Animal Model Bioinformatics Biology Biomedical Research Blood Vessels Cell Culture Techniques Cell Proliferation Cell physiology Cells Chronic Obstructive Pulmonary Disease Collagen Communication Cyst Cystic Lesion Data Deposition Development Diffuse Disease Educational workshop Endothelial Cells Endothelium Environment Exons FRAP1 gene Female Flow Cytometry Future Gene Expression Genes Goals Growth Histologic Human Hyperactivity Inflammation Institution Knockout Mice Knowledge Ligands Lung Lung Lymphangioleiomyomatosis Lung diseases Lymphangioleiomyomatosis Mentors Mentorship Mesenchymal Mesenchyme Methods Molecular Mus Muscle Mutation Pathway interactions Patients Pennsylvania Phenotype Pleural effusion disorder Pneumothorax Proteins Publishing Pulmonary Hypertension Rare Diseases Renal Angiomyolipoma Reporting Research Research Personnel Research Proposals Research Training Resources Role Severities Signal Pathway Signal Transduction Sirolimus Smooth Muscle Stromal Cells TSC1 gene Technical Expertise Testing Therapeutic Intervention Training Transgenic Mice Tuberous Sclerosis Tumor-Suppressor Gene Inactivation Universities Up-Regulation Vascular Endothelial Cell Vascular remodeling Venous WNT Signaling Pathway Work airway obstruction angiogenesis base beta catenin career cell growth cell motility cell type density disease-causing mutation effusion experimental study fitness gain of function human RNA sequencing idiopathic pulmonary fibrosis molecular targeted therapies mouse model myogenesis new therapeutic target next generation sequencing novel pulmonary arterial hypertension pulmonary function decline pulmonary vascular remodeling response single-cell RNA sequencing skills transcriptome transcriptomics triple helix

项目摘要

Project Summary/Abstract Pulmonary lymphangioleiomyomatosis (LAM) is a rare lung disease caused by inactivating mutations in the tuberous sclerosis complex (TSC1/TSC2) gene which results in constitutive activation of the mechanistic target of rapamycin (mTOR) pathway. LAM manifests with diffuse parenchymal cysts complicated by secondary spontaneous pneumothorax, chylous pleural effusions and renal angiomyolipoma. In addition, a subset of LAM patients also develops pulmonary vascular remodeling and pulmonary hypertension. More recently, mTOR has been implicated in other proliferative pulmonary diseases including chronic obstructive lung disease (COPD), idiopathic pulmonary fibrosis (IPF) and pulmonary arterial hypertension (PAH). Despite the growing importance of mTOR activation in pulmonary diseases, the effects of mTOR hyperactivation on pulmonary cells remains incompletely understood. To study the effects of mTOR activation on pulmonary vascular remodeling, I will utilize a novel mouse model with tsc2 deletion as well as single cell RNA sequencing of human LAM lungs. I have found pulmonary vascular remodeling in the tsc2-null mice that mirrors histopathological findings in human LAM. Moreover, our lab recently found that LAM cells hijack the lung transcriptome leading to WNT pathway upregulation in the LAM lung. This proposal will examine how WNT activation in mTOR hyperactivated lungs contributes to pulmonary vascular remodeling with transgenic mice created by the cross of our novel tsc2-null (mTOR activated) mouse with Ctnnb1 (β-catenin) WNT pathway-activated mouse lines. This project will expand my training to include key methods and concepts in cellular crosstalk and signaling. My training will comprise of individualized mentorship, coursework/workshops in flow cytometry, training in bioinformatics and animal models. My training will take place under the sponsorship of Dr. Vera Krymskaya, a leader in LAM research. The Krymskaya lab was the first to establish human LAM cell cultures to demonstrate efficacy of rapamycin for inhibition of mTORC1 and abrogating LAM cell growth. This work will be conducted at the University of Pennsylvania, a world-class research institution with a rich intellectual environment, collaborative investigators and extensive resources for the pursuit of biomedical research. Together, the research and training plans proposed herein will facilitate a better understanding of the pulmonary microenvironment and endothelial-mesenchymal crosstalk while preparing me for my future career as an independent investigator in the field of pulmonary biology.

项目概要/摘要肺淋巴管平滑肌瘤病 (LAM) 是一种罕见的肺部疾病，由淋巴管肌瘤失活突变引起结节性硬化症复合体 (TSC1/TSC2) 基因，导致机械靶标的组成性激活雷帕霉素 (mTOR) 途径的 LAM 表现为弥漫性实质囊肿并发继发性。自发性气胸、乳糜性胸腔积液和肾血管肌脂肪瘤此外，还有 LAM 的一个亚型。最近，mTOR 还导致患者出现肺血管重塑和肺动脉高压。与其他增殖性肺部疾病有关，包括慢性阻塞性肺病（COPD），特发性肺纤维化（IPF）和肺动脉高压（PAH）的重要性日益增加。 mTOR 激活在肺部疾病中的作用，mTOR 过度激活对肺细胞的影响仍然存在不完全理解。为了研究 mTOR 激活对肺血管重塑的影响，我将使用一种新型小鼠我发现了 tsc2 缺失模型以及人类 LAM 肺部的单细胞 RNA 测序。 tsc2 缺失小鼠的血管重塑反映了人类 LAM 的组织病理学发现。实验室最近发现 LAM 细胞劫持肺转录组，导致 LAM 中 WNT 通路上调该提案将研究 mTOR 过度激活的肺部中的 WNT 激活如何促进肺部。通过我们的新型 tsc2-null（mTOR 激活）小鼠杂交产生的转基因小鼠进行血管重塑使用 Ctnnb1（β-连环蛋白）WNT 通路激活的小鼠系。该项目将扩大我的培训范围，包括细胞串扰和信号传导方面的关键方法和概念。我的培训将包括个性化指导、流式细胞术课程/研讨会、流式细胞术培训我的培训将在 Vera Krymskaya 博士的赞助下进行。 Krymskaya 实验室是 LAM 研究的领导者，是第一个建立人类 LAM 细胞培养物来展示的实验室。雷帕霉素抑制 mTORC1 和消除 LAM 细胞生长的功效将在 2017 年进行。宾夕法尼亚大学是一所拥有丰富智力环境的世界一流研究机构，共同致力于生物医学研究的研究人员和广泛的资源。本文提出的研究和培训计划将有助于更好地了解肺微环境和内皮间质串扰，同时为我未来的职业生涯做好准备肺生物学领域的独立研究者。